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300+ Rentals · Milan
300+ Rentals · Milan
Home away from home! That's how Cristina makes you feel. Incredible Thank YouMauriceJanuary 2016
Cristiana's appartmente was beautifull, exactly as pictured. Everything well setup, clean and convenient. She was reacheable at all times and helped us with everything. The place is near the train so you get to downtown milan in 10 mins tops!DamianMay 2016
Nice, clean room close to public transport and very friendly host. I recommend staying here.BobMay 2016
The place was really nice and, most of all, cristiana was an amazing host that provided us with all we needed..NicolandreaJanuary 2016
I had an awesome experience here. Cristiana was very nice and helpful, although we didn't meet personally, she left the keys near the house, and I found them easily with the pictures she sent. The room was very clean, and the neighbourhood was also great. You shouldn't miss this one if you want to live far enough from the centre to have some quiet time. In a word, everything was excellent. Thanks Cristiana!ShuchunMay 2016
Cristiana is nice and the room is prefect!YyJune 2016
Cristiana is absolutely a very nice host! We got much help from her and the house was so lovely and cozy! Very clean and comfortable to live in! It's very convenient to go to the Bovisa station to get around the city! Had a wonderful experience here and recommend it! :DWEIXIDecember 2015
A great flat, with breakfast an drinking in the fridge. Cristina cares good about us and told us some good places in milano. she is very organized :):):)EllaJune 2016
Room Type
Private room
Property Type
Bovisa, Milano
The room is exactly how it looks in the photos - cosy and bright and it really feals like a home for a few dayt that you get to stay out there. Definitely reccomend it :)NeringaMay 2016
Silvestro was the best host we could imagine. He is a kind, friendly and an easy person. If you dont know what to do else in milano don't hesitate to ask him cause he and his friend knows really nice places in milano! We had a quite pleasant stay! Hope to see him again one day ;DTue MinhMarch 2015
We like the interior decoration of this apartment. The host is nice and helpful. The location is very quiet and tourists won't worry about the noise.TIANNovember 2015
Silvestro, thank you for all you did for us! You are very kind person, and we very happy that chose your apartments! Very cozy and cute room! And very tasty tea!)) Also, 10 min from metro by bus or 20 min by foot..)AlikFebruary 2015
Silvestro was really nice. We didn't get to see a lot of him when we were there but he was helpful. The space was comfortable, and easy to get around! He was really accommodating! :)EvaMarch 2015
Loved his flat and had a great stay with Silvestro! The apartment is spotless and the bed was really comfortable (It didn't even feel like a sofa bed). Silvestro was very friendly and communicative throughout our stay. We only stayed for one night, but I wish we could have stayed longer in Milan. Definitely recommend! This is a great airbnb (:ChaminieJuly 2015
We stayed at Silvestro for 4 days and everything was great! The room is really as in the pictures, he let us use the kitchen and he also has a very cozy balcony. The neighborhood is calm and everything is close (supermarket, bakery, caffé, bus stops). The flat is a bit far from the centre but it's quite easy to reach it by bus and metro even at night. Silvestro was very kind and he suggested us some places to visit, which truly worth it.KatalinJune 2016
Until 10 am of our travel day we didn't know at which time we could arrive. (We'Re From austria-travel time 5 h) We didn't meet Silvestro, but big thanks to Valeria who showed us everything. The appartement was ok but there was something really bad smelling in the fridge. But all in one we had some nice days.NiciAugust 2015
Room Type
Private room
Property Type
Baggio, Milano
Room Type
Private room
Property Type
Zona 8, Milano
Room Type
Private room
Property Type
Barona, Milano
- Really good and helpful information concerning the city and the facility (of the host) - Reliability and kindness of the host - Restaurants and stores in the neighborhood are very easy to access - Apartment and furniture are very goodJohannesOctober 2015
Just as described. Loved it.NickJune 2016
The apartment was good, except for the bed, please buy a new bed, the matress is so thin and it feels like you sleeping on the ground. Also the kitchen is not complete with cleaning stuff so we had to buy it.MariekeJune 2016
Andrea and Massimo, have provided all we need . It was wonderful!!! They helped us ( after check out) with our luggage . Everything was correct and all the data were sent in the first few messages. Tks for yout attention and hospitality :) ElizeteElizeteNovember 2015
Very bright and clear apartment! Very beautiful area and good location. Perfect hosts and really perfect Milan!!! What you need more?.. Idial for young peopleIrinaSeptember 2015
I had an amazing stay here! The location is fantastic, and the room is super clean. I would stay here again. Thanks Andrea, wish to see you again in Milano!FanqiOctober 2015
My stay at this apartment was great! Its in a very central location with nice shops and good eating places nearby. The apartment was clean and tidy with everything included. Andrea was waiting for me with the keys when i arrived and he was very helpful. Very good place to stay in Milan !HassanJune 2016
Highly recommend this apartment. It appeared to be what the host has described. It is clean and comfortable, together with a fully-equipped kitchen and a comfortable bed. Also, the apartment is well located, with less than 5 mins walk to both tram and metro station, which allowed me to travel to city center in less than 20 mins. What's more there is a supermarket within 5 mins walk from the apart, which is a nice place to buy some groceries. Although Andrea didnt serve me during my stay, I have a great experience with the guy who served me instead. He gave me a lot of advice on having an amazing journey in Milan. You wont regret it if you stay in these apartment when you travel to Milan!!Chun TingJuly 2016
Room Type
Entire home/apt
Property Type
Zona 8, Milano
Room Type
Entire home/apt
Property Type
Zona 7, Milano
The apartment is really great: well located and well organized. It is perfect for a couple willing to discover Milan! Marcello and Filiberto were great hosts, they give us all the necessary information about the city and were always available for any question. I would recommend it!Claire-MarieSeptember 2014
We had the most wonderful time on our trip to Milan! The Tibaldi Apartment was just perfect for the long weekend that we were there. Picking up the keys was a breeze and the information on how to find the apartment were simple and concise. Filiberto was so speedy with the communication before we arrived and Marcello gave us great information when we picked up the keys! He was extremely friendly and gave us some great suggestions on where to explore. The apartment was spotless and we really loved the simple layout. The location is brilliant, just a simple tram ride into the center and a stroll away from the canals. We had a fantastic weekend and would recommend the apartment to anyone who is taking a short break to explore the city! Thanks again to Filiberto and Marcello! Georgie and Damon :)GeorginaDecember 2014
Everything was perfect !! Great location. Thanks Steph !Mariana February 2016
Tibaldi apartment is really convenient for visiting Milan. We really enjoyed our stay there. The location is perfect. It's only 10min Walk from the subway station, it's located in navigli district that is full of nice bars, restaurants and supermarkets and it's just in front of bus and tramway stations. Furthermore, Filiberto and his colleague gave us all the required information to spend a nice time in the apartment and in Milan. Highly recommended !MounaAugust 2014
Filiberto was a great host. He gave us detailed instructions on how to get to the apartment and once we arrived the placed was light are airy. I dont do not recommend have a few vinos before you attempt the ladders to the bedroom. The location was good for the canals and close to the metro to get into the heart of Milan.EmmaApril 2014
The apartment is wel located, near the nightlife so you can go out every night walking. I have no doubt that location is the best about it. It has a hair dryer, towels and everithing that is necessary for a couple of days. We had a problem with the hot water and Filiberto was easy to contact and the problem was solved in few minutes. But... The apartment is not big as it looks in the pictures. It is not as clean as I think it should be. There is some mold in the bathroom, and ceiling pieces falling into bed. Besides this, everytime that you have to open the door you spent some minutes because the keys does not works as it should.TiagoApril 2016
The apartment is very nice. Small but useful, different and intriguing. You have to use 3 different keys to get into apartment, it is sometimes tiring. Meeting at Office and then going to apartment is also tiring. Meeting must be directly at the apartment not to lose time in a city where you visit first time. When leaving the apartment, it is said to leave the keys in the apartment but you cannot open the doors without keys, so how ? This apartment has some tricks that you should cope with.SaylanNovember 2015
Nice little room, quite close to the channel where a lot happens in Milan. Everything was clean and really similar to the pictures, we had to retrieve the keys at the central station agency and Filiberto's colleague was nice and helping.RaphaëlSeptember 2015
Room Type
Entire home/apt
Property Type
Zona 5, Milan
Oxana was a wonderful host. She made our stay in Milan very comfortable and easy. We were ending our trip at her place and needed to get to the airport very early in the morning. Oxana helped us tremendously with finding a ride and even called us a taxi! She also provides a continental breakfast and maps of the city.CaitlinJune 2016
Sweet, cozy and great organized. The appartment is simple but very clean and perfectly arranged for the guests. You will have complementary sweets and fruits ready for you, amazing real coffee from the coffee machine filter for water, nice and very cozy kitchen to start you day in Milano. The room feels very private so you will not notice anyone living in the appartment. You will be provided with nicely washed and ironed towels and bed sheets, you can find shower gel and shampoo at the shower cabin, and even the new slippers in case you request it. Oxana herself is very nice, very helpful host. She is online almost 24/7, she is advanced (SENSITIVE CONTENTS HIDDEN) user and it feels like she cares about her guests. She is very intelligent and following all the agreement and even more. I consider myself very lucky choosing her as a host and her apartment to stay during my trip in Milano. At her apartment instantly you're feeling like at home. Ludmila (the lady taking care of the apartment) is also very helpful and sweet. The location is about 25 mins walking from the Duomo (main square) and 1.3 km far from the nightlife area and very quite restorante were locals are going. It is a great choice for any occasions and if you don't mind to share sharing the space (buy you won't even notice it). And the most important you are going to feel like a welcome guest not like a customer. Highly recomended! Thank you Oxana for everything!AnnaMarch 2016
Oxana was very friendly with lots of advice about the city. She was always making sure we are ok with everything and very helpful regarding the best transport routes. I would definitely recommend her place!SamJune 2016
My stay at Oxana's place was very pleasant. The apartment is very clean and quiet. It has a very well equipped kitchen. My room was nice with a large, comfortable bed. She also left me some snacks and fruit for my arrival and for breakfast. Even if we didn't get to meet and chat face to face because it was Easter (her roommate waited for me) she was very concerned about my satisfaction and very helpful in answering all my questions. She was also very fast in meeting my requests and confirming my reservation, even if they came at late hours. Thank you for hosting me!CristianMay 2016
In our opinion Oxana's apartment is not more or less than you need for a short trip in a city. We were fully satisfied with the apartment, our room was perfect for sleeping, the bed was comfortable, and it was all clean. The kitchen was full "upgraded", we had a waterboiler, coffee machine, mikrowave, refrigerator and a diningtable in the middle and there were also everything clean and organized. We got from Oxana juice and cookies, she was really nice and helpful. We had wifi connection everywhere in the apartment, and Oxana also provided us with "standard/usual" plugs for electrical devices(only in the kitchen), because the apartment is equipped with italian types. The bathroom is big,we had no problems with it, it was clean, there was a bathcabin and a tub. The apartment has a really good location, 10min to the park, 5-8min to a supermarket, and 20min to the center of the city,also there is a metro station within 8 minutes walk. We had a really good time in Milan and Oxana's apartment was a really good decision.GáborApril 2016
Oksana, thanks for the hospitality. Room: near the center (20 minutes walk), clean, the apartment is very unusual, a lot of beautiful old furniture. Oksana: very attentive to me; on arrival, in the living room waiting for me fruits, sweets, coffee and many other pleasantness :), you can Always ask for help and Oksana always helped. Thank you. I wish You good tidings. :)АлександраMarch 2016
Oxana's place is very friendly it is close to metro and railway station. In the neighborhood are plenty of good restaurants. The apartment was clean and nice. She offered us fruits, coffee and juice, too. I really enjoyed being in Milano, I would go again to Oxana's place.LiviaMarch 2016
Oxana was a extraordinary host. We arrived in the middle of the night but Oxana's friend awaited us with tea and biscuits. We could ask Oxana anything about the city and got a helpful response almost immediately. So we had no problems at all discovering the city. The flat itself is very nicely located. You can walk easily walk to the Duomo in 20-30 minutes and have a supermarket, restaurants and cafes nearly right in front of your door (one restaurant is literally right in front of the door and was just amazing). The kitchen, bed- and bathroom are exactly like shown in the pictures. Nice and cozy. All in all I can say that you can do nothing wrong with staying at Oxana's. Nice flat, perfect location and an extraordinary, very helpful host.ArminApril 2016
Room Type
Private room
Property Type
Zona 8, Milano
The apartment was very modern and cozy. Good for what you are paying for. The assistant Cindy was very friendly and welcoming. Was a little hard to find because the instructions don't really tell you if the house is to the right or left, just says a couple of meters from the subway exit.PamelaFebruary 2016
Everything was preety much as it was listed in preview. Nice,new-renovated apt,close to the center and to Milano fair,with underground station infront of the house.MilanMarch 2016
A very good experience , she was aware of our time arriving , clean , closet for transportation . However ir was a bit far away from downtownMarleneJune 2016
Maria, Sabrina and their assistant Cindy were all very accommodating! The flat looked exactly like it's pictures and was a comfortable stay. ThanksLeoMay 2016
We chose this apartment as our first night's stay in Italy. We only stayed one night and it was the perfect quiet place to rest our heads after a very long and tiring journey from the USA. We were late by 20-30 minutes but Cindy, the lady who let us in was very gracious and welcoming! She showed us where everything is and everything was very clean and neat! We loved how quiet the neighborhood was as well, so we could have a good night's sleep. The apartment is very close to the train station, so that was very convenient, it also made it easy to get to Milano Centrale! The Milan Metropolitano is pretty easy to navigate, and with Maria's directions, finding the apartment is not hard. I loved it even more that she was very accommodating with my questions about Italy (via text) itself and was very helpful! Loved our stay! Would recommend to all of our friends (singles & couples)!StephanieMay 2016
Myself and my partner spent 2 days in Milan and Sabrina could not have been more helpful and flexible. The apartment was easy to find and the metro right downstairs made it so easy to get in and out of the city. The apartment itself was clean and spacious. Complimentary coffee was a big bonus after a long flight too :) Thank you.AmyJune 2016
Sabrina was a terrific host with a lovely apartment to offer, right next to the metro. Highly recommended.HannyJuly 2016
Maria's place was wonderful. It was clean, spacious, and very nicely decorated. WiFi worked well and everything went great. I highly recommend this place to anyone staying in Milan.ForrestJanuary 2016
Room Type
Entire home/apt
Property Type
Affori, Milano
The experience at Chiara's apartment couldn't have been any better. First, she was there to meet us on our arrival and took the time to explain us how everything worked, not only the apartment but also how to move in the city, where to go, etc. It was fantastic! Then, the pictures were more than accurate: when we got there it was even nicer than what it looked like in the pictures! The place is completely renovated, security in the building is great and she lives right across the hallway. Any time we needed some sort of assistance, she was available (for instance, printing boarding passes). Since we stayed like 3 weeks, she even sent a cleaning lady at her own expenses more than once! Regarding apartment location, I can say it's the best spot in the city. 3 blocks away from the Duomo, 1 block away from the famous Monte Napoleone Street, 3 blocks away from Vittorio Emanuele Gallery, and it even has a Taxi Stop and Metro Station right below the apartment. There is no doubt we will go back to Chiara's apartment!LucasSeptember 2015
Chiara greeted us upon arrival. She made great suggestions. The apartment is in a great location...walking distance to metro, duomo, shops, and taxi stand. We utilized her Milan city guidebook. We stayed for 2 nights. Highly recommend staying in this apartment. Chiara is a great hostess.KyleMay 2014
The apartment could not be located in a better place! And as great as the location of the apartment is its hostess, Chiara. She was really nice to us and informed us about lots of interesting things about Milan and helped us deciding where to go. It was a really great experience!SofíaNovember 2013
Great apartment in an ideal location for Milan. Charia was very helpful & accommodating.HelenaApril 2015
Our first time in Milan and our very first time to opt out of luxury hotels on our holidays. Chiara's place was perfect for us as it was closely located to Via Monte Napoleone and Via della spiga, the famous shopping district. It was within easy walking distance to Duomo, the overwhelmingly beautiful structure. If you arrive, you will be surprised by how big Corso Venezia is and by how well organised the surrounding district is. To our surprise, Chiara's air conditioner was a Daikin which is known to be the best air con and it functioned well. Having come from Hong Kong, a hot and humid country, Milan's 37-38 degree summer was bearable but definitely Chiara's place which was naturally cool having been located within the courtyard helped a lot! Chiara was friendly and texted us back as soon as she could when we had some queries. The place looks better than the pictures- she changed the position of the main bed, which was a lot better from our view. Water pressure was good. Warm water came out well. Plenty of hangers. The only downside was that cutleries and plates were not so clean. We didn't cook much- just had a few instant noodles and ate out most of the time. But we felt for those who'd like to make more use of the kitchen might feel disappointed. Apart from it, no major issues. A very comfortable stay. We are happy to recommend Chiara's place to our close friends. Our first air bnb experience turned out to be highly satisfactory. To add extra info, we didn't go to Expo but Chiara's place is literally one minute from metro San Babila on M1 line which is linked to major attractions in Milan. Be ware of tiger mosquitoes in Milan during summer. Chiara's place was safe but outside I got bitten by them. They gave me some hard times. You must put on preventive sprays and once bitten, seek for medical help. Another good thing about Chiara's place is that a pharmacy is near by. We regret we previously had arranged to move to a hotel for our last night. Next time we will stick to Chiara's place throughout our stay in Milan.IndukJuly 2015
Very good experience in Milano thanks to Chiara and her central apartment. 10 mins away from the duomo and very close to everything, pretty clean - perfect for a couple!ThomasFebruary 2014
Chiara's apartment in Central Milan was absolutely amazing! It was exactly as the photos show and more! Everything was well appointed and it had everything I needed for my stay in Milan. It was clean, cosy, modern, close to everything! Had an amazing location with the duomo within 10 minutes walk, corso buenes aires shopping up the road, Emmanuel shopping around the corner which is a nice stroll, restaurants, grocery store, dolce and gabanna martini bar across the street which is nice to have a drink or lunch. The train station is literally 20 metres down from the apartment and so easy to use and get around Milan. It was the perfect location and chiara was a wonderful host. She even found a weight scale for me on my last day and went out of her way to ensure I had it! Thank you so much! Chiara also has a very useful folder with all the essential information you will need for Milan, how to get around, restaurant and cafe recommendations, shopping areas and other districts that are a must see and do. This was very useful and I loved it! Chiara was very quick with responding and was just wonderful! So polite, sweet, nice and easy going. It was an absolute pleasure to meet you and even more of a pleasure staying at your beautiful and wonderful apartment! I would definitely recommend staying at chiara's apartment and I will definitely be back to stay next time I'm in Milan without a doubt!RefikaJune 2014
I recommend Chiara's place to everyone! It is a very clean and cozy apartment and you get exactly what you see. The location is brilliant! I didn't have a problem getting a bus, taxi or the subway. Also the flat's window faces away from the street so it's actually rather quiet despite being a busy neighborhood. Chiara as a person was very helpful with everything and it was super easy to communicate with her As i mentioned before i highly recommend it and i will come back for sure!DIMITRISOctober 2013
Room Type
Entire home/apt
Property Type
Centro Storico, Milan
Great location and house. Great host extremely available for everything I needed.paoloMay 2015
Tutto come da descrizione!paoloNovember 2015
Room Type
Entire home/apt
Property Type
Zona 5, Milano
We had a fantastic break staying in Fabio and Rossana's place. It feels more like an apartment because you have your own private front door out onto the terrace. We met Rossana who was very helpful and always available when contacted. We would definitely recommend this place, and Milan as a place to visit. It really is a beautiful city.GillyMarch 2016
Our stay was brief but the apartment was very clean and comfortable in a nice neighborhood 15 - would recommendSophieFebruary 2016
The place was cosy and the localization was ideal, very close from the center. Rossana welcomed us very nicely, she spoke great french. And Fabio phoned to ask if everything was fine. We left the car around the flat and traveled inside Milan by Metro, which is fast and easy. The only bad thing was that there were no shutters or even dark curtains for the sun. So we had full light in the room at 7 in the morning... not so nice when you are on holidays. Appart from this point, the flat was perfect, very clean and functional. Thanks again for your warm welcome, Rossana ! Elisa & GregelisaMay 2015
The apartment was exactly as described. Rossana was very professional and nice and provided us with city maps and some recommendations on where to go for some drinks and dinner. Also she was very sweet and brought over a pot to make some tea and an umbrella due to rainy weather in Milan. She even let me check in a little earlier so I didn't have to soak outside in the rain.The apartment is just like that of a nice hotel room - stylish, modern, easy to use with all the necessities for a quick trip to the city. Be ready to walk up 5 flights of stairs. It opens up to a very cute Italian style patio where we had our lunch and breakfast so well worth the effort. Fabio, Rossana's brother who we had please to meet, conveniently lives next door in case of any need. The location of the house is excellent - right next to Venezia metro station and near the main shopping street. There are plenty of bars nearby to go for apperetivo and dinner time. We absolutely loved it! Will be back. Thanks Rosanna and Fabio 3aleksandraMay 2015
Fabio & Rossana's place was great. The apartment was just as depicted and was centrally located within Milan. It was easy to get to from Milano Centrale train station (about a 15 minute walk) and the Duomo was only a 20 minute walk. The only negative we'd say was that it was not noted that there would be stairs, 100 steps to the apartment from the ground floor. Luckily, we are young and healthy enough to handle it, but we do believe that it should be noted for those who might have troubles. Other than that, we definitely feel as if we have found a place we would come back to in the future! :)MichaelApril 2016
Very pleasant stay in the apartment. It was exactly what we needed for a couple of nights in Milan as it is walking distance from the centre. The apartment was modern, clean and comfortable with great air con (we needed it!) and also a lovely terrace to relax at the end of the day. We would return to stay here again!BethAugust 2015
Fabio was a great host, very helpful and understanding when our baggage was lost by the airline. The apartment was perfect and very central. The only downside was that it was on the fifth floor with no lift.BrettJuly 2015
The flat was clean with a comfortable bed. It is in a very good area, 20 minutes walking from the Duomo. Rossana was a nice host, she gave us some useful information and in the flat you also have maps and tourist information. I recommend it.AnettaMay 2015
Room Type
Entire home/apt
Property Type
Zona 3, Milano
Lucio is very friendly. I spent one night at his room on Saturday (not-working day). He modified the check-in and check-out time for me. I enjoyed my stay in Milano.AkifumiOctober 2015
I spent just one overnight at Lucio. They are wonderful hosts ! very helpful and caring ! The appartment is in a great Location ! easy walk to all the sights if you´d like. It´s absolutely clean and very charming decorated. I would definitely recommend this place !klausNovember 2015
Lucio has been incredibly friendly and helpful. The house is 10 minutes walking from Central station and 15 minutes walking fron city centre. The flat is very elegant , the cat is adorable, and the pull out sofa confortable (matress extra strong perfect for a very good night sleep). I will definetely go back thereSoniaDecember 2015
Lucio's place was very comfortable and in a great location. I was traveling between Milan and Venice and his place was very close to metro stations that led to Milano Centrale and if you wanted to walk, it was about 15 minutes only. Once I got there, he gave me a few maps and pointed out some ways to get around the city to see all of the main attractions, which came in handy. Him and his partner were very nice and offered to make dinner the first night but sadly, I had already made plans. I'd highly recommend if you're going to Milan for a short stay and want to get around the city's central area.ShanilDecember 2015
It was perfect. the location and his flat at all. best host i ever met on my traveling in Italy. I really appreciate his Kindness and warmly welcoming! that was only one night, but i felt really comfortable and i spent time with Lucio and his friends together. Everything was exactly same as descriptions. because of him i really want to visit milano again. thanks for the perfect day:)IreneJanuary 2016
Lucio is a great host who owns a lovely flat with a cat! He explained me how to get to his place well ahead of my arrival so I could prepare myself well beforehand. His friend was waiting for me although I arrived at the flat around midnight so it was very kind of them. His sofa was very cozy and big enough (for me who's 162cm tall) so I slept very well. I recommend Lucio and his flat!YunjungJanuary 2016
Everything was like in another comments ,when I was reading it. Only thing that I can add.... the place was so easy to find that you never can lose your way to homeAniMay 2016
Lucio was a great host, both prior and upon my arrival. When I booked my stay, he sent me directions and photos on how to get to his apt from the train station. He was very warm and welcoming! The apt was clean and his cat was sweet. Lucio was very nice and invited me to have dinner at his home. He also checked in on me during my trip to make sure all was well and see if I had any questions.BriannaNovember 2015
Room Type
Shared room
Property Type
Centro Storico, Milano
Silvana was a very nice host. We had a breakfast every day. Even when we woke up at 4.30 for our flight back home. The appartement is even better as we thought, perfect clean and very baeutiful view from the big balcony. All in all a perfect host and appartement!! :)TobiasJuly 2016
We had a wonderful stay with Silvana! She was very welcoming and always available to answer questions. She even picked us up when we were dropped off in the wrong location by our taxi. The room and apartment looked just like in the photos and Silvana set up a lovely breakfast for us every morning. The bus/metro system was easy to navigate so getting to the city center was not difficult. I would highly recommend staying here! Thanks Silvana!KailaMarch 2016
She is a SUPER HOST. We were in very emergent situations, and she gave us quick response and was willing to help solving our problems. She provided everything we needed, and she let us have the privacy in a big room with the bathroom. She was also kind to give us pick up at very late night, which we really do appreciate. We had fun talks together, and she provided nice breakfast too! Although it has some distance to reach the center, the bus and metro are easy to reach, and it was enough for us to take a nice tour:) Thanks for all the memories Thanks to our host, we spent wonderful days in Milano, and I recommend this host to all현진July 2016
Silvana was an amazing host. The breakfast was lovely. The room is excellent with a great balcony. Grazie MilleAzSeptember 2015
We loved it at Sissy's. She is an excellent host and we would return. We plan to recommend her to our friends beautiful view from deck and short walk to bus 63 then metro.DavidOctober 2015
Staying at Silvana's was the best experience ever, she is the best host you could find on airbnb. we felt like home since we got there. Always available to help with anything. I definitely super mega recommended her, I'm coming back to Milan in two weeks unfortunately for me her place was booked for those days.ArturoSeptember 2015
Silvana was so sweet! Unfortunately our train got in very late but she waited up for us and let us in and showed us around. She had work early the next morning and even prepared us breakfast before she left. Although we only stayed one night I would stay here again!CourtneyAugust 2015
Although Silvana's English isn't good she tried her best to communicate with us and made us feel at home and the apartment is lovely.JacquesSeptember 2015
Room Type
Private room
Property Type
Zona 7, Milano
Perfect accomodation in nice location and very friendly host. Thomas thank you for all! I recommend you all my friends.RichardMay 2016
The room we rented was very nice! Thomas was helpful and provided us with the basic nessecities and made sure we felt at home :)DavidMay 2016
Although we didn't have the pleasure of meeting Thomas we really enjoyed our brief stay. The room is delightful: well lit, homey, super clean, organized, and we'll decorated! Also a very good located - near the trian station! There's a cute espresso machine and we appreciate all the little details! 5 stars!Becky and OscarJune 2016
The apartment is very nice, super clean and well decorated. Bed is very comfortable. Super well located, 10 minutes from Milano Centrale. Thomas is very helpful. Better than staying at Armani's 5 start Hotel.IvanApril 2016
The host is a polite gentleman, the room is very clean, we can see that the host is a life-style, looked from the style of the room he is supposed to be a designer or work related with fashion, he introduced a lot for us how to use electrical appliances, it is a pity that we only stay in milan for a day. If return to milan next time, I hope we can live in his home.JeanFebruary 2016
Great place to stay in Milan ! Lovely Hosts, very friendly and here to help you whatever you need. The appartement is very well located in the city center and close to any convenient stores, shops and public transportation. The room was quite big, well furnished and super clean. So was the bathroom. Everything was made to make me feel confortable plus the hosts supplied some quick breakfast snacks to start the day which was very appreciated while being here attending a fair. I do recommend this place not only for the material aspect but also for the human experience. I hope they will keep this high level of hosting. Thank you !FrancoisMarch 2016
It's been a great stay with Thomas. Very interesting and kind man. The apartment is nice and sweet. Simple but well designed. The room is clean. It's very nice of him even preparing us breakfast. Recommend.雨晨(Talia)March 2016
the Room is nice, Thomas is kindly helpful. the location is easy to find and near to tbe city center. recommend!MINGFEN May 2016
Room Type
Private room
Property Type
Centro Storico, Milano
Great place All what was offered was provided Exelent place Easy access to public tranport 100 per cent satisfaction Thank you Ilaria for your time and helpMarioNovember 2015
Ilaria is such a sweet and welcoming host! The space is lovely and clean and sunny, and Ilaria is very thoughtful and provides you with towels, toiletries, snacks and even earplugs. The location is a little far from the city centre, but right by the bus and tram stops so it's pretty convenient. Overall my friend and I had a very good time staying here!CaitlinMarch 2015
Extremely helpful host, hip flat with everything we needed and more. Location is a bit tricky though so be sure to follow the host's directions.MikikoMarch 2016
Perfect perfect perfect - regards SteffenSteffenOctober 2015
beautiful room, kitchen very well equipped, bathroom clean. good connection with tram to downtown. very few announcements from train station in the morning but not disturbing if window closed. possibly other guests in the other room. thanks Ilaria :)Tobias July 2013
I'm sorry to say that our experience wasn't as great as that of others. In part, we chose this room precisely because of those comments, but at the end were deeply disappointed. The apartment itself is not bad. However, it isn't exactly in the nicest part of town, nor is it very easy to get there. You have to take the subway and the a tram, and quite frankly, if you don't know the city, it is very easy to miss the stop, which is exactly what happened to us. But our real complaint is the lack of understanding on behalf of the owner, for rather than being comprehensive about our tardiness (although we were in constant communication) she nearly scolded us for being late. We only stayed for one night, and slept quite well. But after we left, we received a very angry text message about not having "fulfilled" her house ( what ever that means) and yet another near scolding experience. In conclusion I would say the apartment is nice and Milan is a beautiful city, but I would not recommend you stay here, but rather search for a more easygoing place.MarianaDecember 2013
Ilaria was lovely!! When we arrived to milan after 26 hours travel there was a cab strike, and Ilaria came to pick us up from a train station to help out which was great of her. When we booked the house we thought we were getting the whole house but there was a few models staying in the other room of the house. Which is fine and they were nice too, but just wasn't expecting it. The train station is just outside the window and can be quite noisy but also interesting. Overall we had a great stay in Milan and Ilaria was a great host!robbieMay 2014
Far away from town!Lyliana January 2016
Room Type
Private room
Property Type
Zona 6, Milan
Silvia was a wonderful host. When we arrived, she welcomed us to Milan, showed us a few essential spots (such as the supermarket), and gave us a quick tour of the apartment. She also showed us where we could find her suggested locations to visit in the Airbnb app. She was available via the app during the entirety of our stay for suggestions and help navigating whatever issues we had, not only with the apartment, but also with activities we wanted to do but could not understand, and always responded very quickly. She also accommodated an early check-out and some minor maintenance that we noticed was needed. The apartment was very clean, cute, bright, and had everything we needed. It was in a quiet residential area that was very accessible to Porta Romana metro and bus station (which allowed for easy transport to Centrale train station and then the airport), a supermarket, and a number of restaurants and bars. We had a great time, and would happily stay again.WilliamJuly 2015
Silvia's apartment was the perfect place for the two of us. It's just as shown on the pictures. It has all you need, it was very clean and very charming. It's located in wonderful neighborhood in Milano; the subway station is a short walk away, and the tram lines are practically in front of the building. There are good restaurants, coffee houses, and supermarkets all around. Silvia is a great and gracious host. I only have praises for her. We'll certainly stay at this place when we come back to MilanoCarlosMay 2016
Silvia is a very nice host. I had a late check-in and an early check-out but was very easy going about it and helpful. The apartment is georgous and well designed. It has air conditioning which was grateful. It also have every thing you need (coffee machine, washing machine, hair dryer) and so much more : candies, fresh flowers and fresh water. The location is great : tramway station in front and a 10 min walk to the subway station. The street is also beautiful with the tram and a line of old trees. My stay in Milan was awesome and I totally recommand this place. I even might come back there !AdelineJune 2015
Silvia was an amazing host. Our train was delayed an hour and she waited for us until our arrival. Silvia went above and beyond by having instructions on almost everything printed out for us, and even offered to demonstrate how things in the apartment worked. The apartment was very well equipped with everything we needed (even water bottles and delicious Italian candy). The apartment was extremely clean. Wifi connection was fast. The location of the apartment was great - less than a 10-minute walk to the nearest metro station, with a few bars and restaurants even on our street. Overall, it was a great stay and I look forward to returning to Milano and staying here again :)KarishmaJune 2015
We had been traveling for 4 weeks and Silvia was the best host we encountered. She has thought of everything. The apartment is very well equipped, from a first aid kit to a cappuccino machine and of course wifi. There is also a little indispensable binder. It has the bus and metro lines, nearby restaurants, nearby shopping, all kinds of great information. I wish all host did this. Silvia made us feel very welcome and went above and beyond for us. She even printed our boarding passes. Five stars for Silvia.DebraOctober 2015
Silvia's flat was really well located (just a few steps for Porta Romana) and very well connected to public transport. The flat itself is in an old building but the interior has been renovated and it's been decorated with lots of taste. It was really clean and had all the necessary for our stay in Milano. Would definitely recommend it. Thanks Silvia for all your support!SaraMay 2016
We were hesitant on deciding whether to use airbnb or a hotel for our trip to Milan and I'm glad we found this listing. There is no way a hotel can match this charming little apartment in any way. The location is perfect, so is the neighborhood, and apartment itself has everything you need. There are also those little touches such as complimentary Nespresso, candies and cookies. How cool is that? But most important of all is the way Silvia takes care of her guess. This is by far the best airbnb experience we've ever had. This is what makes this listing different and restores faith in the service. There is simply no way a hotel will go such an extra mile to make your stay a joyful experience. And did I mention the mini guide waiting for you at the arrival? Well now I did, and it is extremely useful. Thanks for everything!AlexanderNovember 2015
Silvia's apartment was great for our short stay in Milan. It was clean and just as it looks in the pictures. It was very conveniently located close to both the metro and tram stops. Silvia was very quick to respond to my messages and we enjoyed trying a few bars in her neighborhood that she recommended to us. Overall a great experience.KellyJanuary 2016
Room Type
Entire home/apt
Property Type
Centro Storico, Milano
Book without waiting for your reservation to be accepted


text:
Staff was mostly friendly. Rooms were ok.
PROS: Cool Art Deco building with nice veranda, pool area and lawn area. Good location, walking distance to restaurants and shopping.
CONS: It should be beachfront, but there was some sand dredging work being done right in front of the hotel. There was a huge fenced in area with very large and loud equipment – an ugly sight. Not hotel's fault but… There was no hotel beachchair/umbrella service at beach, so hotel referred us to vendor that offered guests a discount. Turns out vendor was set up next to the noisy equipment and they offered ½ off umbrella rentals (a whopping $2.5 off from the $25 for chair/umbrella set). We ended up to walking away quite a distance to get away from the noise.
Then hotel decides to power wash the pool area while guests were lounging at the pool! They made all guests move to one side of pool and closed off the other side. They were very loud and it reeked of gasoline. Now hotel guests couldn't have a nice experience at the hotel beachfront nor the hotel poolside – they should have at least postpone the pool work until beachfront was cleared.
Pool guy wouldn't help us set up poolside lounge chairs which had being stacked out in the lawn area. I asked for help but he didn't, he just stood there and chatted with co-workers while my 84 year old in-law had to get his own chair and struggle to lift it over other guests to get by! We were assigned rooms on main floor. We asked if possible to change rooms on another floor, but receptionist didn't even check – she just said no. The long hallway (main corridor) was really dark and you could hear people in hallway at all hours. TVs didn't work properly and the 'Wi-Fi for AT&T customers only' didn't work at all. Housekeeping didn't clean rooms until very late in the afternoon, even after requesting service. Manager didn't respond to our request to talk with him. It was a very disappointing experience for us.
- Official Description (provided by the hotel):
- Oceanfront in the heart of Miami Beach's Art Deco District and South Beach's finest location. Authentic deco restoration, directed by our owners and embraced by our hand-selected staff providing cordial hospitality. Features 100 Guestrooms: offering all-white bedding, down comforters, Art Deco decor and unique 1930's-inspired alarm clock with safes, bathrobe and iron/ironing board. Services include: heated swimming pool, Verandah Dining Terrace, WiFi, satellite television and Valet parking, rooftop Terrace and Solarium, business center and Fitness Center. ... more less
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- TripAdvisor is proud to partner with Booking.com, BookIt.com, Expedia, Priceline, Hotwire, Cancelon, Hotels.com, Odigeo and Travelocity so you can book your Richmond Hotel reservations with confidence. We help millions of travelers each month to find the perfect hotel for both vacation and business trips, always with the best discounts and special offers.
- Also Known As:
- Richmond Hotel Miami Beach
- Richmond Miami Beach

subset: commoncrawl


text:
Patent Issued for Light Emitting Device Having Wide Beam Angle...By a News Reporter-Staff News Editor at -- A patent by the inventors Chae, Jong Hyeon (Ansan-si, KR); Lee, Chung Hoon (Ansan-si, KR); Suh, Daewoong (Ansan-si, KR); Jang, Jong Min (Ansan-si, KR); Lee, Joon Sup (Ansan-si, KR); Roh, Won Young ... Equities.com, 1 month ago
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- Great number of group executive officers will be visiting Iran, including Chairman Chey Tae-won and CEOs of each subsidiary - To secure business opportunities which will be created in accordance with the president's tour of Iran - Choosing three ...4 Traders, 2 months ago
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text:
|Publication number
|Publication type
|Publication date
|Nov 18, 1969
|Filing date
|Aug 29, 1967
|Priority date
|Sep 13, 1966
|Also published as
|Publication number
|Franklin R Elevitch
|Original Assignee
|Franklin R Elevitch
|Export Citation
|BiBTeX, EndNote, RefMan
|Non-Patent Citations (1), Referenced by (13), Classifications (23)
|External Links: USPTO, USPTO Assignment, Espacenet
THIN FILM APPARATUS AND METHOD FOR ELECTROPHORESIS, CHEMICAL AND BIOCHEMICAL ANALYSES AND THE LIKE Filed Aug. 29, 1967 4 Sheets-Sheet 1 26% 2| I8A I9 22 r 1111111111 n 1 III!!! 4 INVENTOR.
FRANKLIN R. ELEVITCH BY :"fi
TORNEYS 3,479,265 CHEMICAL Nov. 18. 1969 F. R. ELEVITCH THIN FILM APPARATUS AND METHOD FOR ELECTROPHORESIS,
AND BIOCHEMICAL ANALYSES AND THE LIKE 4 Sheets-Sheet Filed Aug. 29, 1967 GAMMA GLOBULINS ALBUMIN 1 a 9 L 2 e xw 2 U (Cr .32: k B ni h vv w .56.. G A T l E B s m L U B m 9 "'1' 3 G A W H i| P L A 7 3 INVENTOR.
FRANKLIN R. ELEVITCH A ORNEYS Nov. 18. 1969 THIN FILM APPARATUS AND METHOD FOR ELECT F. R. ELEVITCH ROPHORESIS, CHEMICAL AND BIOCHEMICAL ANALYSES AND THE LIKE 4 Sheets-Sheet 3 Filed Aug. 29, 1967 so (64 I I E. El
FRANKLIN R. ELEVITCH TTORNEYS Nov. 18. 1969 F. R. ELEVITCH PPARATUS AND METHOD FOR ELECTROPHORESIS v 3,479,265 CHEMICAL THIN FILM A AND BIOCHEMICAL ANALYSES AND THE LIKE Filed Aug. 29, 1967 4 Sheets-Sheet 4 Its-4|] \l \il "Er-1E1 INVENTOR.
FRANKLIN R. ELEVITCH BY 7%/ ATTORNEYS United States Patent 3,479,265 THIN FILM APPARATUS AND METHOD FOR ELECTROPHORESIS, CHEMICAL AND BIO- CHEMICAL ANALYSES AND THE LIKE Franklin R. Elevitch, 476 F erne Drive, Palo Alto, Calif. 94306 Continuation-impart of applications Ser. No. 300,341, Aug. 6, 1963, and Ser. No. 579,089, Sept. 13, 1966. This application Aug. 29, 1967, Ser. No. 664,133
Int. Cl. B01k /00 US. Cl. 204-480 11 Claims ABSTRACT OF THE DISCLOSURE The apparatus employs a thin film of permeable gel into which chemical reagents or biological fluids such as blood serum and proteins can diffuse. The film has a thinness of not more than about one millimeter or less and it is characterized by uniformity of thickness such that voltage gradients and current densities through the film are uniform when it is used as an electrophoresis medium and such that diffusion of chemical reagents or of biological fluids into the film occurs uniformly. The thinness of the film allows rapid dissipation of heat when the apparatus is employed for electrophoresis and it economizes on the use of chemical or biological reagents which are incorporated in the film. The apparatus may be supplied by the manufacturer to the user as a two-piece mold having a first section to which the gel is adherent and a second section to which the gel is non-adherent. When the customer is ready to use the film, the first mold section (to which the thin film of gel is adherent) is stripped from the second section, thus exposing the thin film supported on the first mold section. This film may be formed with wells for the introduction of reagents, unknown specimens, or the like.
This application is a continuation-in-part of my copending application Ser. No. 300,341 filed Aug. 6, 1963, now abandoned, entitled Thin Layer Gel Electrophorectic Couvette and Method of Electrophoresis and Ser. No. 579,089, filed Sept. 13, 1966, now abandoned, entitled Method for Molding a Film of Gel and Method of Analyzing Electrophoretically Separated Materials in the Gel.
The present invention, like that of Ser. No. 300,341, relates to the use of thin films of permeable gel-like material for electrophoresis and it employs a simplified type of apparatus for providing such films as described in Ser. No. 579,089. The thin uniform films of the apparatus of the aforesaid patent applications and of the present application permit the use of high voltage gradients, for example volts per centimeter and consequent rapid electrophoresis, but without damaging heat buildup and without the need for positive cooling. Heretofore, thick films have been used and, as a result, low voltage gradients have been required (and long periods of time have been required) or positive cooling has been required. The uniformity of thickness of these films provides the advantage of uniform voltage gradients and current densities and consequent reproducibility of results. With thick films as used heretofore, variations of thickness of the order of, say, 0.1 mm. have not been significant, but with films of about 0.5 mm. thickness, such deviations are proportionately much greater and more serious.
In my application Ser. No. 579,089 a simplified apparatus is described which has certain advantages over the earlier apparatus of Ser. No. 300,341; e.g., it is less expensive to manufacture and it lends itself more effectively to manufacture of the entire apparatus including the film at a factory and it avoids the need to form the film at the place of use.
The present application relates to disposable, factory made, thin film equipment of the character described in my application Ser. No. 579,089 and it relates also to improvements upon and variants of that equipment.
Certain forms of the apparatus of the present invention are illustrated by ways of example in the accompanying drawings in which:
FIGURE 1 is an isometric view of a mold which embodies the invention, and showing the cover removed from the base of the mold.
FIGURE 2 is a sectional view taken on line 22 of FIGURE 1 but showing the base of the mold with the cover thereon.
FIGURE 3 is a sectional view taken on line 3-3 of FIGURE 1.
FIGURE 4 is an isometric view showing the gel casting on the base of the mold with the mold cover removed therefrom.
FIGURE 5 is a plan view of a single portion of the gel containing a single set of wells and showing the relative positioning of electrophoretically separated material thereon.
FIGURE 6 is an isometric view which is similar to FIGURE 4 but showing the application of a liquid reagent to the gel during a novel process of accurately measuring isoenzymes.
FIGURE 7 is an isometric view which is similar to FIGURE 6 but showing the liquid reagent being spread across the gel.
FIGURE 8 is a perspective view similar to that of FIGURE 1 but of a different type of mold which produces a thin, permeable film of gel as in FIGURE 4 but with certain modifications which adapt it for use in the radial diifusion-precipitin technique.
FIGURE 9 is a cross sectional view, taken along the line 9-9 of FIGURE 8, but with the mold sections assembled.
FIGURE 10 is a plan view of the film produced by the apparatus of FIGURES 8 and 9 supported on one of the mold sections.
FIGURE 11 is a section taken along the line 1111 of FIGURE 10.
FIGURE 12 is a fragmentary plan view of the film of FIGURE 10 showing how it is used in the radial diffusionprecipitin technique.
FIGURE 13 is a diagrammatic view showing another embodiment of the invention.
Reference is now made to FIGURE 1 of the drawings wherein one form of novel mold of the invention is shown comprising upper and lower portions or sections 10 and 11, respectively. The lower mold section 11 simply comprises a flat member of generally rectangular shape made of any suitable material such as a transparent high optical clarity, low water-absorbent, low-fluorescent, preferably flexible plastic. The upper section 10 which is formed of semi-rigid, transparent, 10w water-absorbent, low fluorescent plastic is provided with vertical end walls 12 and side walls 13 to define a rectangular shaped recess 14 in the lower face thereof. Plastic such as polystyrene is suitable for use in forming the mold sections. The base section 11 is of less thickness than the cover 10 whereby the base 11 is relatively flexible and the cover is relatively rigid. The low-fluorescent quality of the plastic base 11 results in a device which may be used in fiuorometric analysis, and the high optical clarity thereof enables density measurements with a densitometer. The face 16 of the top mold section 10 is adapted to be placed on and secured to the plane upper face 17 of the bottom mold section 11. The upper and lower sections and 11 are made of substantially the same length and width for easy alignment of the cover 10 on the base 11. Although any suitable means may be used for securing the cover on the base, a non-hardenable water-resistant adhesive 18 is shown. (For purposes of illustration, the adhesive is shown only in the enlarged cross-sectional view of FIGURE 3 and not in the other drawing figures.) As described hereinbelow the adhesive is of the type permitting subsequent removal of the cover from the base. In practice, a rubber-base cement has been found to be satisfactory. Obviously, other suitable cements or means such as pressure sensitive adhesives or heat sealing may be used if desired. With the top mold section 10 attached to the lower mold section 11 a rectangular-shaped cavity or chamber 18A is formed by the recess 14 in the cover 10 and upper face 17 of the base 11 as shown in FIGURE 2.
The top mold section or cover 10 is provided with a plurality of horizontally-spaced, downwardly depending dividing ribs 19, each of uniform distance from the next, all parallel with the end walls 12, and of the same height as the end and side walls 12 and 13. Each of the ribs 19 has its longitudinal ends at a fixed distance from side walls 13 thereby forming channels or passageways 21 and 22 at each end of the ribs 19. The position of the ribs 19 creates a series of uniform sized parallel spaces 23 between each of the ribs, and between the end walls 12 and adjacent ribs.
Well-making ribs 24a and 24b (of identical shape but provided with different reference characters for purpose of description) are also formed in the recess 14 of the cover, which ribs are of lesser height than the dividing ribs 19. For example, they may be approximately onehalf the height of the dividing ribs. The well-making ribs 24a and 24b are spaced longitudinally in pairs, and in the illustrated embodiment, one well-making rib 24a of each pair thereof is located in the passageway 21 and the other well-making rib 24b of each pair is formed within one of the parallel spaces 23. In a modification of the invention, the ribs 24a also could be spaced to fit within the parallel spaces 23.
In use, a suitable release agent such as a silicone compound (not shown) is applied to the recess 14 of the top section 10, and a suitable adhesive 18 is applied to the face 16 of the mold cover 10. The cover 10 (to which the gel adheres by capillarity) is then placed on the mold base 11 with the face 16 and the dividing ribs 19 in engagement with the base 11. The cover and base are preferably clamped together to assure the formation of a cavity 18A of uniform thickness.
It is important that the layer of gel formed by the apparatus be uniform in thickness. The base 11 is flexible, hence subject to distortion and the more rigid cover 10 may depart from flatness because of limitation of manufacturing techniques. However, the end walls 12, the side walls 13 and the ribs 19 are of uniform height. Therefore when the apparatus is placed in a press the mold cavity 18a, hence the layer or film of gel produced in this cavity, have a very uniform thickness.
The cover 10 is provided with filling and air vent passageways or openings 26 for filling the mold, which openings may be closed by caps 27. The mold is filled with a suitable medium for electrophoresis such as aqueous solutions of agar, agarose, starch, acrylamide, or the like, through one of the openings 26, which medium except acrylamide is introduced as a heated liquid but assumes a solid form when cooled. Acrylamide is introduced at room temperature and is polymerized in the mold by use of a catalyst such as light or a chemical catalyst. Such solutions are well known. The solution passes along the channels 21 and 22 and between the ribs 19 to fill the entire mold cavity 18A including the spaces 23. The solution is then hardened to form an electrophoresis gel (shown in FIGURE 4 and identified by the reference character 28). The caps 27 are applied to the mold for storage of the gel in the mold until ready for use.
Prior to filling the mold, the recess 14 and the ribs 19 and 24 formed in the recess are coated with a suitable release agent. No release agent is applied to the base 11. After the mold is filled and the gel is hardened, the flexible base 11 with the gel 28 adhered thereto simply is peeled off the cover for use. A plurality of uniformly spaced parallel rows of wells 29a and 29b are formed in the gel 28 by the well-making ribs 24a and 24b, respectively. The wells 29a and 29b are extremely thin, on the order of 0002:0001 inch in width, and are about 0.006i0.001 inch in depth. In one embodiment a gel layer of about 0015:0001 inch thick has been used. A thin gel layer is desired to permit use of relatively high voltage gradients, of about 15 volts per centimeter, without destructive heat build-up. The wells are filled by means of thin capillary tubes with a measured amount of material, each well being filled by capillary action to its volume, and with a uniform amount.
The gel 28 is also formed with a plurality of parallel slots 31 which extend through the film, which slots are formed by the dividing ribs 19. These slots facilitate cutting the gel into uniformly sized strips for examination following the electrophoresis process. Also, the slots 31 may be filled with antiserum when the gel is employed in an immunoelectrophoresis process. As best seen in FIGURE 1, the lower edges of the ribs 19 are curved for line contact with the face 17 of the base, whereby the slots 31 formed in the gel by the ribs 19 have arcuate shaped walls. The walls of these slots 31 are readily wettable by fluid introduced therein whereby the slots are readily filled. The plastic base 11 is not readily wettable and if the ribs 19 were formed with parallel sides, the resultant slots in the gel would be diflicult, if not impossible, to fill. In addition, the ribs 24a, 24b and 19 are all formed with curved edges rather than sharp edges to facilitate removal of the top mold section from the bottom mold section without damaging the gel.
In use, the film of gel 28 on the flexible plastic base 11 is placed on a flat surface such as a desk, counter top, or the like, for loading of the wells 29a and 29b. Biological fluid to be analyzed such as blood serum, is placed in a well or wells. If fewer than eight samples are to be analyzed, the gel and base may be cut, as by a scissors, to the desired length along one of the slots 31, and the unused portion thereof stored in a plastic wrapper until ready for use.
With the well, or wells, loaded, the gel 28 on the base 11 is then placed face down on buffer soaked wicks in an electrophoresis cell of an electrophoresis apparatus. Electrophoresis apparatus which is commercially available and of well known design may be used. Such apparatus, not shown, comprises an electrophoresis cell and a power supply connected thereto. A typical electrophoresis cell is divided into four chambers. Two central chambers have platinum wires running down their centers, and are connected to the outside chambers by cotton or glass wool wicks or filter paper. Opposite edges of the gel film 28, along the long sides thereof, are placed on wicks which dip into the outside chambers of the cell. The power supply is connected to the platinum wires for current flow through the chambers and the gel film 28 thereon.
It is common knowledge that almost all of the inorganic or organic ions in solution which might be placed in the slots or wells will have different mobilities; mobility being dependent upon electrical charge, molecular weight, size, shape, and the like, with electrical charge being the primary factor in electrophoresis. Because the molecules or class of molecules in the inorganic or organic solutions used at any particular time have different mobilities, the molecules of blood serum or virus, or bacteria, as the case may be, will thereby be caused to move a varying distance toward a positive or negative pole of the gel from their starting place in the slots or wells 29a and 2911. In
the illustrated arrangement the one edge portion designated 33 of the gel 28 is connected to the power supply anode, and the opposite edge portion designated 35 is connected to the cathode. The electrophoretic separation is preferably performed at a given voltage gradient (e.g., -15 volts/centimeter) with a constant current over a predetermined time interval,
One method of electrophoretic analysis which involves the use of both of the wells 29a and 29b is described in my above-identified copending patent application Serial No. 300,341. In the method described in said copending patent application, biological fluid to be analyzed, such as blood serum, is placed in the Wells 2912 and a suitable antigen solution, such as a bacteria or virus-containing solution, is placed in the adjacent well 29a. The fluid and antigen are subjected to electrophoresis whereupon the fluid is electrophoretically separated for isolation of the antibodies in the gamma globulin fraction. The antigen reacts with the antibodies to form a complex which may be analyzed. For details of this method, reference is made to said copending patent application.
A novel method of quantitative determination of isoenzymes will now be described employing the gel molded by the novel mold of this invention. The method contemplated by this invention employs a sucrose-agarose gel comprising a 10% sucrose, 1% concentration of agarose, solution in a buffered electrolyte solution made up of sodium barbital and hydrochloric acid, pH 8.8, 0.05 M although other butfers of similar pH and ionic strength may be used if the electrolytes are non-fluorescent. This solution is used ecause it is convenient to make and has no autofluorescence. It will be understood that this gel is of the preferred type, but that the method also may be employed with other gels such as starch, agar, acrylamide, or the like.
In use, biological fluid to be analyzed such as blood serum, is placed in a well or wells 2%. A precise amount of such fluid, e.g., one microliter, is placed in the wells. As mentioned above, if fewer than eight samples are to be analyzed the film and base may be cut to desired size.
With the well or wells 2% loaded, the gel 28 on the base 11 is placed in an electrophoresis apparatus for electrophoretic separation of the biological fluid.
An example of a strip of agarose gel after removal from the electrophoresis cell and cutting into strips and showing the distribution of the now separated biological sample into fractions is shown in FIGURE 5. The albumin is shown at 37, the alpha globulins at 39, the beta globulins at 41 and the gamma globulins at 43. Isoenzymes will lie equally spaced between the remote ends of bands 39 and 43, It will be understood, of course, that after the electrophoresis process has taken place, the tracings shown in FIGURE 5 are not visible to the eye unless some coloring or fluorescent solution, or the like, has been added.
The present method contemplates diffusing a reagent, or the like, in solid, liquid or gaseous form through the thin layer of gel to react with the electrophoretically separated sample. The chemical reaction products are then analyzed in the gel. Depending upon the reagent employed, and the reaction obtained, the analysis is by fluorometric, spectrophotometric, radioisotopic, or the like, means. The present method is not limited to the use of any particular reagent, or to any particular chemical reaction between the reagent and electrophoretically separated sample, or to any particular analysis method or apparatus employed in the analysis of the chemical reaction products. However, for purposes of explanation, one embodiment of the novel method is illustrated in the drawings and will be described in detail. In accordance with this one embodiment, fluorescent patterns are developed for precisely and accurately measuring lactate dehydrogenase (LDH) isoenzymes. After electrophoresis of the biological sample, such as blood serum, in the manner described the LDH isoenzymes are incubated with a reagent. A suitable reagent comprises a substrate or a substrate combined with a coenzyme. An example of the latter comprises sodium lactate and the coenzyme, nicotinamide adenine dinucleotide (NA-D+), which reagent is simply prepared by mixing the lactate substrate fluid with the coenzyrne NAD+. About twenty drops of the liquid reagent are dispensed along the anode side of the electrophoresis gel 28 as shown in FIGURE 6. In FIG- URE 6, the liquid reagent 47 is shown being dispensed from a bottle 49 onto the gel 28. The reagent is then spread evenly over the gel. For this purpose, a glass rod 51, or the like, may be employed as shown in FIGURE 7. The rod is placed on the gel at the anode side thereof and pushed across the film of gel 28 in a single smooth motion in the direction of the arrows 53 without pressing down thereon. A film of the liquid reagent is thereby evenly spread over the gel and the excess liquid swept off the opposite edge of the gel.
The LDH isoenzymes are incubated for ten minutes with the reagent by placing the gel on a support above water level in a covered water bath at 37 C., during which time the sodium lactate is converted to pyruvate and the non-fluorescent NAD+ is converted to reduced nicotinamide adenine dinucleotide which is fluorescent. For determination of total LDH, one or more of the otherwise unused outer wells 29a in the gel 28 may be loaded with one microliter of NADH (l mg./ml.). Preferably, three such wells are loaded with NADH. The gel 28 is then dried in an oven at about 90 C. for approximately fifteen minutes. Because of variations in oven convection and ambient humidity, the drying time is empirically determined. In any event, the agarose film must not make contact with hot metal, and preferably, the gel 28 on the base 11 is placed gel side up on several layers of paper towel or filter paper during drying.
The developed LDH isoenzyme sites appear as bluewhite fluorescent lines under near ultraviolet light (365 my), and the resulting enzymatic reaction products may be analyzed by fluorescence. The gel 28 on the base 11 is cut into individual strips with a scissors, or the like, along the slots 31. The individual strips are analyzed by use of a fluorometer. Preferably, the fluorometer output is connected to a linear recorder for recording the LDH isoenzyme patterns. The percentage of LDH isoenzyme at each site is determined by measuring the peak height of each recorded isoenzyme, and calculating the percent LDH isoenzyme at each site by the following formula:
LDH isoenzyme percent:
Peak height LDH isoenzyme Sum of the peak heights The total LDH isoenzyme present in each strip is calculated by use of the following formula:
Total LDH (in micromoles NADH/ ml./l0min.)=
Sum of the peak heightsxconstant Peak height of 1 g. NADH standard The peak heights of 1 mg. NADH standard is determined by analysis of the wells loaded with one microliter of NADH (1 mg./ml.) as described above. It should be noted that, at the present time, different lots of NADH vary in their fluorescent efliciency. This foregoing formula is satisfactory if the NADH is uniform in this respect, but if it is not, each lot may be calibrated against a uniform fluorescent standard and appropriate corrections may be made.
The above-described method of accurately quantitatively measuring LDH isoenzymes is much simpler than prior art methods. With my novel method the film of reagent spread over the thin electrophoretic gel rapidly diffuses therethrough and is uniformly distributed therein for complete chemical reaction with the electrophoretically separated serum enzymes. The novel method provides for rapid and accurate analysis of the serum enzymes and isoenzymes. It will be understood that the novel determination of isoenzymes of lactate dehydrogenase does not depend upon the electrophoresis of an antigen solution with the serum to be analyzed as described in the above-mentioned copending patent. That is, the serum enzyme analysis may be made on serum which has been electrophoretically separated without electrophoresis of an adjacent antigen solution. In this case, the wells 29a along the edge of the gel 28 would not be used during the electrophoresis process and the NADH (1 mg./ml.) may be placed in these outer wells in the course of performing the novel serum enzyme analysis described above.
Referring now to FIGURES 8 to 12, and preliminarily to FIGURE 8, an apparatus is there shown which is generally similar to that of FIGURE 1 and is designated by the reference numeral 60 and which is a two-part mold as in FIGURES l and 2 but with certain modifications. The mold sections are designated as 61 and 62 and they may be made of the same material as the mold sections 11 and 12 shown in FIGURE 1. Preferably, although not necessarily, the mold section 62 is rigid and the mold section 61 is flexible, such relative rigidity and flexibility being suitably accomplished by making the section 62 of relatively thick plastic material and the top section 61 of relatively thin plastic material. The rigid mold section 62 is formed with a rim 63 which extends entirely around the periphery and which defines the thickness of mold cavity 64. The rigid mold section 62 is also formed with upwardly extending projections or risers 67 and with one or more spacer ribs 66. The ribs 66 have the same height as the rim 63, i.e., they are equal to the depth of the mold cavity 64, and they provide additional support for the flexible mold section 61 and ensure the production of a film of uniform thickness. Alternatively to or in addition to the spacer ribs, some of the risers 67 (described below) may extend to the plane of the upper edge or rim 63 and perform the same function.
In the selection of materials of construction of the mold parts, that is the mold sections and 11 of FIG- URE 1 and the mold sections 61 and 62 of FIGURE 8, certain desiderata are set forth above. It is in order to amplify these desiderata.
The mold section 10 (FIGURE 1) and 62 (FIGURE 8)that is, those parts of the apparatus which define the mold cavity and which, in most embodiments of the invention, also form sample-receiving wells, etc.are preferably rigid and they may be opaque or transparent. The optical properties of these mold parts are not as important as the optical properties of the opposing mold sections 11 and 61 in certain cases because the sections 10 and 62 are discarded at the time of use. However, it is preferred that the mold sections 10 and 62 be transparent because that enables one to inspect the apparatus when filled with gel to determine whether there has been complete filling of the mold cavity with the gel.
With respect to the mold sections 11 and 61 (that is, the sections which are flexible and which are stripped from the other sections with a film of gel adhering to them) optical properties are more important but will vary according to other factors.
Thus, if the apparatus is to be used in a fluorometer having a drum to which the film of gel and its supporting base are attached, it is preferred that the mold section 11 be quite flexible so as to be easily bent around the cylindrical surface but with fluorometric or other optical reading devices which use specimens in flat condition flexibility is not as important, although some flexibility is preferred in any case to facilitate peeling the mold section 11 or 61 from the mold section 10 or 62. With regard to optical properties, transparent material may be used and is preferred in certain situations. For example, if in the measuring technique that is employed, light passes through the film of gel and its base to a detector such as a photocell,
the base supporting the film should be transparent. Where, however, the measuring technique does not require passage of light through the base, for example, in some types of fluorometry and in radio-isotopic measurements, the radiation that is measured is emitted radiation and an opaque background is permissible and may, in fact, be preferred.
Referring to FIGURE 9, the mold sections 61 and 62 are assembled as shown and a gel, e.g., any of those mentioned above in connection with FIGURES 1 to 7, is poured in through one of the holes 68. Displaced air is vented through the other hole 68. The mold cavity 64 is completely filled. The mold apparatus may be placed in a press and allowed to cool to harden the gel.
The adjacent surface of flexible mold section 61 (hereinater referred to as the base 61) is adherent to the gel whereas the inner surface of the rigid mold section 62 including the ribs 66 and risers 67 are non-adherent, e.g., by reason of being coated with a release agent such as a silicone. Therefore the rigid mold section can be stripped from the gel after it has hardened.
Referring now to FIGURES l0 and 11, the film of gel formed on and adhering to the base 61 is indicated by the reference numeral 75. Wells 76 are formed in this gel, such wells being cylindrical and of uniform diameter. Slot 77 is formed by rib 66. As shown in FIGURE 11, each of the wells 76 is separated by a thin layer of film 78 from the base 61. The apparatus of FIGURE 8 is designed so that the film is quite thin, e.g., less than 1 mm. and preferably about 0.3 to 0.5 mm., and it is designed so that this thickness is quite uniform.
The apparatus shown in FIGURES 10 and ll can be employed to great advantage in the radial diffusionprecipitin technique. This technique is described by Stiehm and Fudenberg in a paper entitled Serum Levels of Immune Globulins in Health and Disease: A Survey, published in Pediatrics, vol. 37, No. 5, May 1966, pages 715 to 727. As there explained, a glass plate was coated with a uniform, two millimeter thick-layer of agar-antibody mixture. The antibody used by these authors was a rabbit antiserum to human q/G, M, and 7A globulins. After the layer of gel had hardened, wells were punched in it and uniform samples of abnormal serum under investigation were placed in some of these wells. A standard serum was placed in other of the wells, The plates were submerged in mineral oil for six days at 37 C. and were then rinsed and stained. The staining resulted in visible rings resulting from the precipitin reaction between the specific antiserum and antigen. The areas of these rings were directly proportional to the concentration of antigen inserted into the respective wells. By inserting known quantities of antigen into certain of the wells, and by measuring the ring diameter resulting from known and unknown quantities of antigen, the unknown can be determined.
Thus, radial diffusion of a suitable material such as an antigen from a circular well into a permeable film of gel which is impregnated with a suitable reactant such as an antiserum, results in the well known precipitin reaction and the formation of a ring which is visible even without staining. For example such rings can be observed and measured in oblique light. By adding known quantities of antigen in certain of the wells, followed by measurement of ring diameters, a quantitative determination of the unknown can be made.
Referring now to FIGURE 12, each of the four wells 76 is indicated by a numeral enclosed in parentheses. The precipitin ring around each well is indicated by the broken circle 80, and the area of the annulus 81 between each well 76 and its precipitin circle 80 is a quantitative measure of the amount of active component in the sample introduced in the respective Well. For example; assume that well No. (1) is the well to which the unknown has been added and that wells Nos. (2), (3) and (4) are those to which known but different quantities of standard have been added. Using a calibration curve and by comparison of the areas of annulus 81 surrounding well No. (l) with the areas of annuli 81 surrounding wells Nos. (2), (3) and (4), the amount of active component in the sample placed in well No. (1) can be readily determined.
As stated above, the film of gel 75 is very thin, for example about 0.4 mm. in thickness, and it is also quite uniform in thickness. Uniformity of thickness is, of course, important; otherwise the results would not be reproduceable. The extreme thinness of the film is advantageous because, among other reasons, material such as anti-sera incorporated into the film to react with standard and unknown specimens introduced into the wells are oftentimes very expensive. Inasmuch as all or a large part of the film must be uniformly impregnated with antiserum or other reagent, it follows that a film of, say, two millimeters thickness will require much more of this expensive material than a film which is four-tenths of a millimeter in thickness.
Another feature of importance is the fact that the wells 76 do not reach entirely to base 61 but are separated from it by a thin layer 78 of the gel. This is advantageous because if the wells penetrate to the adjacent surface of the base 61, a certain, not insignificant portion of the specimen introduced into the well will creep under the film of gel rather than diffusing into the gel, thereby destroying or impairing the reproducibility and significance of the results obtained.
Another embodiment of the invention is shown in FIG- URE 13. In this embodiment a length of tape 85 is provided which travels continuously or by increments in the direction shown. This tape consists of a base 61 like that in FIGURES and 11 but in the form of a length of tape. Adhering to the base 61 is a film 75 of permeable gel. This gel, which may be as in FIGURES 10 and 11, may be applied in any manner, e.g., by brushing, squirting, spraying or dipping. It is, however, a thin film as in the case of FIGURES 10 and 11, i.e., preferably not more than about 0.3-0.5 mm. and it is uniform in thickness. Such uniformity can be achieved by various means, e.g., by passing the tape between rollers that are set apart a predetermined distance and which squeeze out excess gel, or a doctor blade may be employed to remove excess gel.
The layer of gel 75 may, if desired, have sample receiving wells formed in it such as the wells 76 in FIGURES 10 to 12. However, there need be no such wells.
The film of gel 75 may be uniformly impregnated with a reagent e.g., by premixing the gel and reagent before applying it to the base 61. The tape is moved beneath a pipette 91 which applies a measured amount of the unknown. If the tape is moved slowly, or if the pipette is caused to undergo a cycle in which it moves forwardly with and at the rate as the tape and moves quickly rear- Wardly to its starting position, the tape may move continuously. Alternatively the tape may be moved by increments and the unknown added during each pause between increments of forward movement.
Then the tape may be processed (after it has travelled a sufficient distance or has paused long enough for the reaction between reagent and unknown to be completed) in any suitable manner. For example, in the analysis of serum protein, the serum protein is applied by the pipette 91 and any suitable dyestulf such as 8-anilino naphthalenel-sulfonic acid is applied by means of a nozzle 92. The selected dyestuff is one like S-anilino naphthalene-l-sulfonic acid which will attach itself to the protein undergoing analysis and in so doing will become fluorescent in proportion to the amount of protein. By this means the amount of protein (the reagent dyestuif being added in excess) can be determined by fluorometry. Other dyestuffs than 8-anilino naphthalene-l-sulfonic acid may, of course, be used provided they function in this manner, i.e., they are non-fluorescent but attach themselves to protein and become fluorescent in proportion to the amount of proteindyestuif complex. In the case of 8-anilino naphthalene-lsulfonic acid, it is preferably used in the form of an acid solution, e.g., 30 mgs. of the dyestutf in ml. of 5-10% aqueous acetic acid solution.
If desired a fixing step may be interposed between pipette 91 and nozzle 92 to fix the protein; e.g., a dilute aqueous solution of formaldehyde and perchloric acid may be sprayed onto the are to which serum protein has been applied.
After the reagent has been applied by nozzle 92, after a fixing step (if any) has been carried out and after such period of time as is required for reaction of reagent with the protein to be completed, the tape is passed through a fluorometer 97 which may be provided with a readout to record the value of the determination on a printed tape and/ or on a screen.
8-anilino naphthalene -1-sulfonic acid is unstable in aqueous solution if held for any great length of time; it should be used when freshly prepared. This leads to another embodiment of the invention, as follows: A film 75 is provided in the form of a tape as in FIGURE 13, or in any other form in accordance with the present invention. 8-anilino naphthalene-l-sulfonic acid is stable for long periods of time if it is dry. The film 75 is impregnated with this reagent which is then dried, e.g., by application of mild heat and/or vacuum. The reagent is reactivated when moistened, which may be accomplished by dipping the film in or spraying it with water just before use. In many cases the water in the sample of unknown, e.g., blood serum, will serve the same purpose. In general, permeable films, preferably but not necessarily made in accordance with the present invention, may be impregnated with a labile reagent which is stable in anhydrous environment but which is unstable in the presence of moisture, and this film may be dried; stored, shipped and delivered in dry form; and reactivated by adding moisture at or just before the time of use.
Rather than using a tape, a disc may be employed having a base as at 61 and an adhering film of gel, and this disc may be rotated in a turret type of apparatus wherein reagent and/ or unknown may be applied at different points and a sector of the disc may rotate through a fiuorometer.
In the tape and disc embodiments of the invention the extreme thinness and uniformity of the film of gel are advantageous not only for the reasons previously stated but because the steps involved, moistening, drying, curing, etc. can be accomplished very quickly. If thick layers of gel are employed, the time required for these phenomena may be prohibitive.
In general optimum thickness and optimum uniformity can be determined by criteria such as those mentioned immediately above and elsewhere above. I have found that thickness less than about 1 mm. should be used, preferably not more than about 0.5 mm.; and that departures from average thickness preferably do not exceed about 10%, most advantageously not by more than about 7%.
It will therefore be apparent that novel and very useful analytical apparatus has been provided.
I claim:
1. A disposable film package adapted for electrophoresis comprising:
(a) an impermeable substantially flat base,
(b) a thin film of permeable aqueous gel having one surface in adherent contct with said base,
(0) cover means in contact with the other surface of said film of gel and in edge contact with the base completely enclosing and sealing the film, and
((1) said film of gel being adherent to said base and non-adherent to said cover means such that when the base is stripped from the cover the film of gel remains adherent to the base.
2. A package according to claim 1 wherein the base is flexible.
3. A package according to claim 1 wherein both the film and the base are transparent.
4. A package according to claim 1 wherein both the film and the base are non-fluorescent.
5. A package according to claim 1 wherein both the film and the base are transparent and non-fluorescent.
6. A package according to claim 1 wherein the cover is provided with well forming projections extending into the film of gel.
7. A package according to claim 6 wherein said well forming projections terminate short of the base.
8. A package according to claim 1 wherein the edge sealing is accomplished by means of a non-hardenable water-resistant adhesive.
9. A disposable film package adapted for electrophoresis comprising:
(a) an impermeable substantially fiat base,
(b) a thin film of permeable aqueous gel having one surface in adherent contact with said base,
(0) cover means provided with projecting spacer members and well forming members on one surface thereof,
(d) said base and cover being assembled with filr'n of gel in between and with the spacer members and well forming members projecting into said film of gel, said spacer members being in contact with said base, and said well forming members terminating short of said base, said cover and base being in edge sealing contact around their entire periphery and thereby completely enclosing the film, said film of gel being adherent to said base and non-adherent to said cover such that when the base is stripped from the cover the film of gel remains adherent to the base.
10. A package according to claim 9 wherein said base is flexible and both the base and the gel are transparent and non-fluorescent.
11. A method of forming a film of permeable gel 12 adapted for electrophoresis comprising:
(a) providing a flat impermeable base,
(b) providing a cover for said base having spacer members,
(c) assembling said base and cover in edge sealing contact with said spacer member projecting toward said base contacting the same and spacing the inner surfaces of the base and cover apart to form a mold cavity, and
(d) filling said mold cavity with a permeable aqueous gel, said gel being adherent to said base and nonadherent to said cover such that when the base is stripped from the cover the film of gel remains adherent to the base.
References Cited Bodman, Chromatographic and Electrophoretic Techniques, Agar Gel, Starch Block Starch Gel & Sponge Rubber Electrophoresis, vol. II, 204-180g. October 1960, pp. 97-105.
Raymond, Clinical Chemistry A Convenient Apparatus for Vertical Gel Electrophoresis, vol. 8, No. 5, October 1962, pp. 445-464, 469-470, 204180g.
Raymond et al., Analytical Biochemistry, Preparation and Properties of Acrylamide Gel for use in Electrophoresis, January 1960, pp. 391-396, 204-180g.
Smithies, Journal of Biochemistry, An Improved Procedure of Starch Gel Electrophoresis, vol. 71, No. 3, 1955, pp. 585-587, 204180g.
JOHN H. MACK, Primary Examiner E. ZAGARELLA, JR., Assistant Examiner US. Cl. X.R. 204-299
|Citing Patent
|Filing date
|Publication date
|Oct 28, 1970
|Jul 4, 1972
|Millipore Corp
|Electrophoretic apparatus
|Dec 27, 1974
|Jun 22, 1976
|Medac Gesellschaft Fur Klinische Spezialpraparate Mbh
|Chamber and process for crossed immunoelectro-phoresis
|May 24, 1977
|Sep 26, 1978
|Corning Glass Works
|Electrophoretic analyzer with automatic reference circuit
|Jul 1, 1980
|Feb 9, 1982
|Beckman Instruments, Inc.
|Electrophoretic gel container
|May 28, 1980
|Feb 16, 1982
|Karlson Eskil L
|Apparatus for continuous electrochromatographic separation
|Nov 14, 1986
|Dec 1, 1987
|Helena Laboratories Corporation
|Container for an electrophoretic support medium
|Jan 5, 1987
|Jul 26, 1988
|Helena Laboratories Corporation
|Container for an electrophoretic support medium
|May 20, 1987
|Jan 3, 1989
|Bios Corporation
|Method and apparatus for molding thin gel slabs horizontally with integrally molded large volume sample wells
|Jul 17, 1987
|Jan 9, 1990
|Helena Laboratories Corporation
|Electrophoresis plate and method of making same
|Feb 22, 1989
|Dec 4, 1990
|Helena Laboratories Corporation
|Electrophoresis plate and method of making same
|Apr 21, 1995
|Apr 30, 1996
|E. I. Du Pont De Nemours And Company
|Gel cassette for enhanced electrophoretic separation and processes for the preparation thereof
|Oct 4, 1995
|Aug 6, 1996
|Biotech Holdings, Inc.
|Gel electrophoresis cassette
|Jul 1, 1981
|Jan 6, 1982
|Beckman Instruments, Inc.
|Electrophoretic gel container
|U.S. Classification
|204/620, 422/940, 422/945
|International Classification
|Cooperative Classification
|European Classification


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