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Showing results for tags 'Samsung'.
Found 30 results
blurry Album art is blurry on lock screen (Samsung, Android 9)
Oscar H posted a topic in Bug ReportsThe lock screen feature on my Samsung A50 (Android 9) doesn't work. Cover art shows blurry, even though the album art is well embeded. Happens with every song. I tap the player on the lock screen and it show the album art, which is blurry. Album art properly shown. Lock screen. You have to tap the feature to show the album art. Album art blurry.
Distorted/weak/low sound after Android Pie update
Lando posted a topic in Bug ReportsI recently updated my Samsung Galaxy S8 software to a new version of Android Pie and once I launched the Poweramp app I noticed the sound was distorted and low. It sounded like it had a DVC slapped on it, so once I noticed this I checked the settings and they were the same, the DVC was off. But when I switched it off and on there was the slightest difference. I've been finding others with the same issue as far back as January from the forums and I just hope this can get fixed and update can get rolled out on the app as soon as possible since I'm assuming the app isn't compatible/ not functioning with this version of Android Pie.
support Samsung's ultra powersave
kostas posted a topic in Feature RequestsCould you support ultra power save on Samsung devices? (android 9.0 s9+) (you need to select apps that u want to use in ultra powersave and Poweramp is not listed)
Hi-res Output:sample rate 384khz
Meandr posted a topic in Feature RequestsSamsung galaxy s9 +:adapt sound and Hi-res Output:sample rate 384khz can't work at the same time. (But 192khz can )
Excess battery consumption
RBEmerson posted a topic in General Poweramp ChatterIt appears that PA is a serious battery eater, cycle burner. Starting from about 90% charge, the battery on my rooted (otherwise stock ROM, and Andy 7.0) Sammy S7 is close to flat within about 5 hours. This is using plugged in Klipsch earbuds .In addition battery consumption, the phone gets noticeably hot (no warm). High battery consumption and hot phone all point to a very busy CPU. What's going on here?
Hi! i updated my Samsung Galaxy S9+ to Android Pie on 15th December and when I played music the sound was way too low, usually perfect sound on 70% but not after update, before update, the Samsung Alive equalization never interfere with Poweramp but now is it, help me, I tried everything like turning off dvc but nothing, sorry for my bad English
Hey, soo yeah as the title suggests, i have problems with my s8 and Poweramp after i upgraded to 9.0. The volume is considerably lower, the lower end is weaker, and Poweramp is considered as a cast device (i think, will include photos). When i turn the volume, instead of the "Media" volume bar popping up, a separate bar with a cast icon appears, and has no warning past the 9 level, suggesting it's recognised as a speaker or something? I'm not sure, i'm not entirely sure i understand it. When i disable DVC, the volume corrects but the sound is distorted and the lows bleed into the mids. Same happens if i disable Hi-Res output (48khz/16bit, i read in another post it has to be that). I tried denying Soundalive permission to change system settings and even tried to force stop it, along with Poweramp. I'm not sure what mught be causing the issue, i messaged samsung and they suggested i contact the developer. I submitted a bug report, but i'm posting here aswell in case someone has a solution. Here are the links to the screenshots: https://drive.google.com/file/d/1g_eV7m5CdBOM3PpcF50ZTJQ7SVkR8THk/view?usp=drivesdk https://drive.google.com/file/d/1SVjwg7zq6SSVrNaOReLqvXIGNkUb0HaG/view?usp=drivesdk Thanks in advance!
Samsung Note 8 - Bluetooth resume after phone call
bchliu posted a topic in Bug ReportsCurrent Android version on the Galaxy Note 8. Playing music over Bluetooth and get a phone call. Phone works fine and answer call. Once call is complete, Poweramp resumes the music with the phone speaker instead of through bluetooth once again. Happens on all of my Bluetooth devices from my Bose Headphones to my car audio system.
Output Lag
Myksharma posted a topic in Bug ReportsAfter updating to latest version of Poweramp, it start lagging in audio output. I use sound assistant, both system controller & Sound assistant controller are fine in other apps. When I adjust volume from any controller output cuts some time. Volume is 60/150 but output is like 10. My version is V3 build 816.
Edge Panel
va9rant posted a topic in Feature RequestsHi Max! Love the new app! Will it be possible to incorporate a feature or add-on for Samsung Edge panel?
Samsung and DVC
moyu posted a topic in General Poweramp ChatterHello Samsung S8 Exynos variant still get problem : in audio setting DVC is said to be disabled by ROM by actually it is enabled. I went to audio output setting and use Open SL ES Hi-res output and turn on disable DVC for it and then DVC is disabled finally. I don't hate DVC. To be honest I love DVC but on Samsung Oreo seems that DVC is not compatible with system. When DVC is turned on the sound becomes very small, can't hear it with normal volume. I need very high volume to make music to be able to hear. Do you have this problem? Thanks.
안드로이드 스마트폰 마우스 스크롤 문제 android smartphone mouse scroll issue
기명5 posted a topic in Korean/한글 포럼안드로이드 스마트폰에 마우스를 연결해서 사용하려고 하는데(예를 들면, 삼성 덱스), 파워앰프 애플리케이션에선 마우스 스크롤이 작동되질 않더군요. 이 문제 해결해주시기 바랍니다. 수정) 파워앰프 베타 버전의 경우 I wanna use android smartphone with mouse(for example, Samsung Dex), in Poweramp application, mouse's scroll does't work. Please solve this problem. Thank you. Edit)In case of Poweramp beta version
Poweramp version: 2.0.10-build-588-play (Full Version) Device model: Sprint Samsung Galaxy S5 (SM-G900P) Android version: Oreo (8.1.0) ROM: LineageOS (version 15.1-20180723-NIGHTLY-kltedv) I noticed that after a while, some FLAC files on my phone start getting corrupted. I could tell because it would start always skipping at at the same exact point in a song. I confirmed that the files were corrupted when I copied them back on my computer and did the "flac -t *.flac" command using the flac.exe command-line tool. Maybe it's a bad microSD card (I'm using a genuine SanDisk Extreme PLUS 128 GB formatted as non-portable storage), but I doubt it since I believe I had this issue on my Extreme PLUS 32 GB card as well. I think my pictures and videos are fine and uncorrupted as well (I don't think I have the time to check them all, though). Anyone else here ever have a similar issue? Is it a Poweramp issue or is it a bad microSD card? Maybe it's an issue with Android or LineageOS? Thanks.
Hey guys! I recently got a new phone (Samsung Galaxy S8), and metadata like artist name, album name, song title, etc no longer displays on my car (2012 Mazda 3)'s heads-up display. It has worked perfectly fine before this on my last phone (LG G4). Music plays just fine, it's just that the metadata doesn't display. I realize this is most likely an issue with the phone itself using a different version of bluetooth compared to my last phone, but since you guys are the gurus when it comes to troubleshooting this kind of stuff... is there any chance you can look into it, and see if it'd be an easy fix to implement in Poweramp (v2 or v3, I've tried it on both) itself? Like, maybe a toggle-able option to make Poweramp force the phone to transmit metadata the old way so that it'll display in situations like this? Just an idea. Not sure who else to contact/ask.
I have the last official version 2.0.10 build 588 play version in my Samsung Galaxy S8 Plus (SM.G955FD) with Android 7.0 and the option Triple double press for next/last song doesn´t work with my headphones. I also have a Moto G 3 smartphone and it works perfectly there.Why is it happening? I'm so sad because i have the paid version and i can not find answer
Samsung S8 High Res Audio support
biffwick posted a topic in Feature RequestsI was wondering when power amp is going to add high res flac support for the Samsung galaxy s8 as it has high res playback
i am using Poweramp v2 build 588. Android ver Nougat 7.0 Stock Samsung rom rooted. I created my playlists in stock samsung music player and i tried to import them to Poweramp but Poweramp says imported playlists = 0 i also cleared Poweramp data and uninstall it but all in vain it still says 0 entries. What could be the problem.? Please Help
Lockscreen popping up 2-3 times
rafa posted a topic in General Poweramp ChatterHi there, I use Poweramp pro for .. 4 years now and I am really happy with the product. great stuff! however, since I got a new phone, a samsung galaxy s7, I am seeing an issue with the lockscreen. whenever I click the "to lock screen" button on the Poweramp lockscreen, it switches back to the phone lock screen for a second or two and then the Poweramp lockscreen appears again. this is really annoying since it not only happens when going to the phone lock screen, but also when accessing the camera of the phone by samsungs lock screen shortcut of double tapping home and not having to go through the finger print/unlock procedure. To be fair, I think this issue existed before the beta, but I found out now with the new phone and thought the new release would be an option to fix the bug.. Cheers, rafa
Hi, I cannot diagnose why Poweramp is skipping/hiccuping/stuttering. I am using a Samsung Galaxy S4 and have tried it on stock Touchwiz 5.0.1 and 4.4.2, Poweramp 2.0.10 build 588 I am aware of the article http://support.powerampapp.com/knowledgebase/articles/323527-skips-hiccups-stutters And the results from trying to use this have are: Not an SD card issue (music plays perfectly in stock player) Not Bluetooth issues (using regular wired headphones) Not tweaked ROM (I use stock Touchwiz 4.4.2 and 5.0.1, unless Touchwiz is considered tweaked?) No additional 3rd party audio effects (Adapt Sound has never been touched/set up) No CPU hungry apps (Show CPU usage in developer options does not indicate high CPU usage and GSam battery monitor shows the highest battery usage being Android OS itself) No broken formats (skipping is not consistent with particular files) If you could please help me resolve this problem I would greatly appreciate it. I have reset my phone on stock 4.4.2 and 5.0.1 in attempts to resolve it.
Need help. Bluetooth tag output issues
AimEgoGod posted a topic in General Poweramp ChatterI recently upgraded to an S7 and put all my music on my new phone (old was an S5.) I used Poweramp premium on both phones and I listen to my music primarily in my car via bluetooth. When I had my S5, the tag info would pop up on my 2014 Civic's display. With my S7, it shows no info. When I play songs in google play the tags show up but not when using Poweramp. Does anyone know how to fix this issues? Thanks
Carlhefler posted a topic in General Poweramp ChatterUsing power amp beta via Bluetooth. At incoming call, music briefly comes out phone and when call disconnected, blue tooth not resumed. Power Amp freezes.Bluetooth
Galaxy EDGE Widgets
swillbur posted a topic in Feature RequestsBAck, Play/pause, Forward.......and lyrics showing when phone is turned off with either a galaxy s6 edge or a note edge.
After syncing songs from my laptop, many songs are randomly disappearing from my Poweramp library. I've tried reloading multiple times but songs are still disappearing. The song files still appear in the Google Play Music app so I know they've been successfully loaded on the Sandisk micro SD my Samsung Galaxy S5. Is anyone else having this issue? If so, were you able to correct this?
Air view & air gesture support on sgs4
Koblavi posted a topic in Feature RequestsIt would be nice if you could support these features on the s4. Skip tracks using air gesture. Preview track/album information using air view. Briefly preview songs when your finger remains on it.
Star Ratings & S4 Adaptive Sound
rexmcartor posted a topic in Feature RequestsI have to two requests for future features. 1. This I have asked for many many times via E-mail. I was told the first time it was planned to be added in the next update (but it still hasn't and has been well over a year). I would LOVE it if you would make it so Poweramp can read the Ratings tag already in all my files. I have every single song I have rated and would love for Poweramp to be able to read these, and not make me re-rate them. Also, standard rating includes 1/2 star (making it out of 10 instead of 5) which is much better. 2. The second request. The Samsung S4 has a feature called Adaptive Sound. This plays tones and then equalizes your left and right headphones to sound more balanced based on your ears. I absolutely love this feature, due to a damaged right ear. I have asked for this feature to be added to Poweramp before but no response of it. Now Samsung did the work of making this feature. Now can you please make Poweramp use it. IDK why but it seems to only work in the default player app (which sucks and I paid for Poweramp). I have tried changing all the setting available in Poweramp but it still doesn't seem to work with adaptive sound. Please, please, please make it work. I will be using the standard music app until then.


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Not good qualities for a parrot seeking a new home.
Still, honesty is the best policy, so the Royal Society for the Prevention of Cruelty to Animals has decided to come clean about his shortcomings.
Supervisor Angelina Lusher said Wednesday that anyone who takes Beaky in will have to put up with some rude language. The agency also says he tends to bite people until he bonds with them.
Beaky is a chattering lory who will only be given to new owners with experience handling exotic birds.
Lusher says Beaky needs interaction with people or other birds to have a fulfilling life.

subset: commoncrawl


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Vandy Vape Ni80 Superfine MTL Fused Clapton Wire 32GA*2 38GA - 10ft - 5.35ohm/ft
Resistances wire Vandy Vape Ni80 Superfine MTL Fused Clapton Wire 32GA*2 38GA.
Contact atmi-zo.gr
Secure transactions
Fast shipping
Returns accepted (According to Terms and Conditions)
Τα προϊόντα με την ένδειξη "Τιμή Internet" αφορούν προσφορές αποκλειστικά μέσω ηλεκτρονικής παραγγελίας. Δεν είναι δυνατή η αγορά τους σε φυσικό κατάστημα στην τιμή προσφοράς.
- Product type: Resistance wire
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- Resistance: 5.35ohm/ft
- Length: 10ft (3.048m)
- 1 x VandyVape Ni80 Superfine MTL Clapton Wire 32ga*2+38ga - 10ft
All orders are checked by the store before they start for any defects. Our products are of the highest quality and we wish to have completely satisfied customers.
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Intel P67 (B3) chipset based
• Military Class II Concept components: Top quality & stability
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• THX TruStudio PRO: deliver the fullest audio experience
• IGP Supports DirectX 10.1, Shader Model 4.0 and OpenGL 2.0
• Support USB 3.0 & SATA 6Gb/s
Military Class II Concept
The perfect trinity of quality, stability and efficiency all exist at one motherboard to meet end user expectations. The MSI Military Class II series products provide the long lifespan, unparalleled quality, extreme performance and ultimate efficiency.
OC Genie II
Just press the OC Genie button or enable OC Genie II function in BIOS menu, the CPU performance (P67 series) or the 3D performance of iGPU (H67 series) will be highly improved. Whether users want to watch a high bit-rate HD movie or go online gaming, both can be enjoyed with image performance that is smoother and lag-free!
APS (Active Phase Switching)
Active Phase Switching, APS, technology is an intelligent design that helps to save energy on MSI's motherboards. The underlying idea is to switch off the power supply as soon as no electricity is needed, and when the system needs more resources again, APS will automatically adjust electricity supply accordingly. APS will automatically detect your processor's loading at any given time and save energy accordingly. This is only possible by means of a very advanced power controlling technology. Other than software-based solutions, APS technology stands out from MSI's own research. The specially developed IC chip can operate automatically depending on the demands of the power supply, which is to reduce the total power usage.
B3 Stepping Ready
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Parsnips are a quintessential cold weather vegetable. They look like carrots but are white and somewhat sweeter, characteristically starchy with a hint of citrus.
Parsnips are a quintessential cold weather vegetable. They look like carrots but are white and somewhat sweeter, characteristically starchy with a hint of citrus.
Like many of the tastiest foodstuffs, parsnips were once a staple among the poor in Italy and elsewhere in Europe. When the potato came from the New World and disseminated throughout the continent, parsnips fell out of favor. For centuries, they were used almost exclusively as animal feed.
Parsnips are complex in flavor, remarkably versatile and easy to cook. Their earthiness works well in broths and soups but also comes through when roasted or braised.
I like to slice parsnips thinly and cook them with rendered pancetta for a simultaneously crisp and toothsome pasta sauce. Don't crisp the slivers like potato chips, just cook all the way through and toss with spaghetti and a few ladles of pasta water.
I even serve parsnips instead of mashed potatoes as a Thanksgiving side, adding Yukon gold potatoes and yams. The combination of sweet potato and parsnip might seem like it would be too sweet, but it creates the perfect balance.
This dish works next to a turkey or with poultry or braised meet for weekday meals. It beautifully captures the flavors of autumn.
Parsnips with Horseradish and Chives
Serves 8 to 10 as a side dish. Recipe courtesy of "Molto Batali" (ecco 2011).
3 pounds parsnips, peeled and cut into 2 1/2-inch-long batonettes
1/4 cup plus 4 teaspoons extra virgin olive oil
Salt and freshly ground black pepper
1 cup prosecco or champagne
6 tablespoons unsalted butter, at room temperature
1/4 cup plus 1 cup prepared white horseradish
1/4 cup finely chopped fresh Italian parsley
1/4 cup chopped fresh chives
2 tablespoons poppy seeds
Preheat the oven to 400 F. In a large roasting pan, toss the parsnips with the 4 teaspoons olive oil and salt and pepper to taste. Add the prosecco, and stir. Place the pan in the oven and roast, stirring once or twice, until the parsnips are tender and the prosecco has been absorbed, 25 to 30 minutes. Meanwhile, heat the butter and remaining 1/4 cup oil in a small saucepan over medium heat. When the butter has melted, remove from the heat and stir in 1/4 cup of the horseradish. Add the parsley, chives and poppy seeds, and season with salt and pepper. Pour the horseradish-herb butter over the warm roasted parsnips, and toss to coat. Serve in a warmed shallow bowl with the remaining 1 cup horseradish on the side.
Mario Batali is the owner of Babbo, Lupa, Otto and other renowned restaurants. His latest book is "Molto Batali," published by Ecco.)


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Substrates, systems and methods for analyzing materials
Substrates, systems and methods for analyzing materials that include waveguide arrays disposed upon or within the substrate such that evanescent fields emanating from the waveguides illuminate materials disposed upon or proximal to the surface of the substrate, permitting analysis of such materials. The substrates, systems and methods are used in a variety of analytical operations, including, inter alia, nucleic acid analysis, including hybridization and sequencing analyses, cellular analyses and other molecular analyses.
Latest Pacific Biosciences of California, Inc. Patents:
- De novo diploid genome assembly and haplotype sequence reconstruction
- Labeled nucleotide analogs, reaction mixtures, and methods and systems for sequencing
- Hierarchical genome assembly method using single long insert library
- Arrays of integrated analytical devices and methods for production
- Modular nucleotide compositions and uses therefor
This application a continuation U.S. patent application Ser. No. 11/849,157 filed Aug. 31, 2007, which claims priority to Provisional U.S. Patent Application No. 60/841,897, filed Sep. 1, 2006, the full disclosure of which is incorporated herein by reference for all purposes.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH
Not applicable.BACKGROUND OF THE INVENTION
A large number of analytical operations benefit from the simultaneous illumination of relatively large area of substrates in order to accomplish the desired analysis. For example, interrogation of biopolymer array substrates typically employs wide area illumination, e.g., in a linearized beam, flood or reciprocating spot operation. Such illumination allows interrogation of larger numbers of analytical features, e.g., molecule groups, in order to analyze the interaction of such molecule groups with a sample applied to the array.
In the case of DNA arrays, in particular, large numbers of oligonucleotide probes are provided in discrete locations on a planar substrate surface, such that the surface comprises multiple, small patches of identical probes, where the probes' nucleotide sequence for each patch location is known. When one applies a labeled sample sequence to the array, the position on the array to which the sample hybridizes is indicative of the complementary probe sequence, and as such, the sequence of the sample sequence. These arrays are generally interrogated using laser based fluorescence microscopes that are capable of applying excitation illumination over large areas of the substrate in order to interrogate all of the patches. Such systems have employed galvo scanners, slower, scanning microscopes, linearized beam illumination, and wide area flood illumination.
In some cases, however, a more tightly controlled illumination strategy may be desired. For example, it may be desirable to provide stricter control of the volume of material that is illuminated, as well as the overall area that is illuminated, effectively controlling illumination not only in one of the x or y axes of a planar substrate, but also in the z axis, extending away from the substrate. One example of controlled illumination that accomplishes both lateral (x and y) and volume (z) control is the use of zero mode waveguides as a base substrate for analyzing materials. See, U.S. Pat. Nos. 6,991,726 and 7,013,054, the full disclosures of which are hereby incorporated herein by reference in their entirety for all purposes. Briefly, zero mode waveguide array substrates employ an opaque cladding layer, e.g., aluminum, chromium, or the like, deposited over a transparent substrate layer, and through which are disposed a series of apertures through to the transparent layer. Because the apertures are of sufficiently small cross sectional dimensions, e.g., on the order of 50-200 nm in cross section, they prevent propagation of light through them that is below a cut-off frequency. While some light will enter the aperture or core, its intensity decays exponentially as a function of the distance from the aperture's opening. As a result, a very small volume of the core is actually illuminated with a relevant level of light. Such ZMW arrays have been illuminated using a number of the methods described herein, including spot illumination, flood illumination and line illumination (using a linearized beam) (See, e.g., co-pending U.S. patent application Ser. No. 11/483,413 (filed Jul. 5, 2006), and 60/772,908 (filed Feb. 13, 2006), the full disclosures of which are incorporated herein by reference in their entirety for all purposes).
While the various foregoing systems and methods have proven some measure of effectiveness, the present invention provides for improvements over these systems and methods, in a number of respects.BRIEF SUMMARY OF THE INVENTION
The present invention provides substrates, systems and methods for analyzing materials. In particular, in at least one aspect, the invention provides an analytical device that comprises a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface. The device includes an analyte disposed sufficiently proximal to the first surface and external to the waveguide, to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide, e.g., the light field that decays exponentially as a function of distance from the waveguide surface.
In another aspect, the invention provides a method of illuminating an analyte that comprises providing a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface. An analyte is provided disposed sufficiently proximal to the first surface and external to the waveguide, to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide. Light is then directed or propagated through the first waveguide such that the evanescent field from the waveguide illuminates the analyte.
In another aspect, the invention provides a system for analyzing an analyte that comprises a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface. At least a first light source is provided optically coupled to the at least first waveguide to direct light into the first waveguide. An optical detection system is also provided positioned to receive and detect optical signals from an analyte disposed sufficiently proximal to the first surface and external to the waveguide, to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide.
The present invention generally provides a simplified method for delivering illumination to a plurality of discrete analytical regions on a substrate, and does so in a manner that provides better control of that illumination. In particular, the present invention provides methods, systems and substrates that include surface exposed waveguides, such the exponential decay of light outside the waveguide may be exploited in the surface region in the same manner as the light that enters a ZMW, above, in order to selectively illuminate materials provided upon that surface.
The invention provides such substrates alone, as well as in conjunction with one or more other components in an overall system, such as reagents (e.g., dyes, enzymes, buffers and other adjuncts, and the like), illumination sources (e.g., lasers, lamps and the like), observation and detection components or systems (e.g., optical trains including lenses, mirrors, prisms, gratings and the like, detectors such as PMTs, Photodiodes, photodiode arrays, CCDs, EMCCDs, ICCDs, photographic films, and the like).
Also provided are methods and applications of the substrates and systems of the invention in the illumination and preferably the analysis of materials which more preferably include fluorescent or fluorogenic materials. As will be appreciated, the present invention is broadly applicable to any application in which one desires to illuminate materials that are at or proximal to a surface and/or specific locations on a surface, without illuminating materials that are not similarly situated. Examples of such analyses include illumination, observation and/or analysis of surface bound cells, proteins, nucleic acids or other molecules of interest.
In the context of analysis, the substrates, methods and systems of the invention impart numerous additional advantages to an analytical operation. In particular, because the light of interest is applied in a spatially focused manner, e.g., confined in at least one lateral and one orthogonal dimension, using efficient optical systems, e.g., fiber optics and waveguides, it provides a much more efficient use of illumination, e.g., laser, power. In addition, because illumination is provided from within confined regions of the substrate itself, issues of illumination of background or non-relevant regions, e.g., illumination of non-relevant materials in solutions, auto-fluorescence of substrates and/or other materials, reflection of illumination radiation, are substantially reduced. Likewise, this aspect of the invention provides an ability to perform many homogenous assays for which it would be generally applicable.
In addition to mitigating autofluorescence of the substrate materials, the systems described herein substantially mitigate auto-fluorescence associated with the optical train. In particular, in typical fluorescence spectroscopy, the excitation light is typically directed at the reaction of interest through at least a portion of the same optical train used to collect the fluorescence, e.g., the objective and other components. As such, autofluorescence of such components will contribute to the detected fluorescence level and provide a fair amount of noise in the overall detection. Because the systems direct excitation light into the substrate through a different path, e.g., through an optical fiber optically coupled to the waveguide in the substrate, this auto-fluorescence source is eliminated.
Other advantages include an automatic alignment of illumination with analytes of interest, as such alignment will be self defining, e.g., an analyte of interest may be that which is provided within a pre-existing illumination region. This level of alignment is also designed (and is thus programmable) and fabricated into the substrate and is thus not prone to any misalignment issues or other sensitivities of other analytical systems. As such, the alignment of illumination is highly robust.
Finally, the substrates of the invention typically are provided from rugged materials, e.g., glass, quartz or polymeric materials that have demonstrated longevity in harsh environments, e.g., extremes of cold, heat, chemical compositions, e.g., high salt, acidic or basic environments, vacuum and zero gravity. As such, they provide rugged capabilities for a wide range of applications.
Such illumination is particularly useful in illuminating fluorescent and/or fluorogenic materials upon or proximal to the surface, including nucleic acid array based methods, substrate coupled nucleic acid sequencing by synthesis methods, antibody/antigen interactions, and a variety of other applications. These and other applications are described below.II. Substrates
In particularly preferred aspects, a waveguide substrate is used that provides for the propagation of a single beam and preferably its separation into multiple propagated beams in a waveguide array. Examples of such arrayed waveguides have been employed in the fiber optic based systems in splitting beams propagated by a single fiber into multiple fibers. Typically, however, such arrayed waveguides employ a cladding layer disposed over the waveguide carrying substrate. An example of such a device is schematically illustrated in
In contrast to the above described waveguide arrays, the present invention exploits the underlying waveguide substrate and waveguides, but eliminates the cladding layer that blocks access to the light that emanates from the waveguide, thus exposing the substrate surface and providing more proximal access to the evanescent light field emanating from the waveguides at the substrate surface. As noted previously, a large number of analytical operations can benefit from the ability to controllably illuminate materials at or near a surface and/or at a number of locations thereon, without excessively illuminating the surrounding environment. Further, although discussed in preferred aspects as providing access to the evanescent light from an array, it will be appreciated that modifications of waveguides to enhance direction of light propagating therethrough, toward the surface, are also contemplated by the invention, including, e.g., embedded gratings or other optical components embedded in the waveguide that direct light toward the surface region of interest on a substrate. Notwithstanding the foregoing, exploitation of the evanescent field is preferred as it gives rise to a desired limited illumination volume proximal to the surface of the substrate for a number of applications.
An example of the devices (also referred to herein as analytical devices) of the invention is schematically illustrated in
In the absence of a cladding layer, e.g., cladding layer 150 in
As noted previously, this provides access to the evanescent light outside of each waveguide core. By providing materials at or proximal to the surface, e.g., particle 224, one can controllably illuminate such materials without illuminating any materials outside of the evanescent field. Such selective illumination allows for illumination of individual or relatively small numbers of particles, molecules or cells in more concentrated solutions of such materials, as described in greater detail below. As will be appreciated, controllable illumination includes not only control of illumination in the axis orthogonal to the substrate or core surface (z axis), but also in at least one of the axes of the plane of the substrate surface (x or y axes). This lateral control of illumination, particularly when coupled with additional lateral control of analyte location, e.g., through the use of immobilization, or other spatial confinement techniques, e.g., structural barriers, etc., provides additional advantages of selectivity of observation.
In accordance with the present invention, the substrates may include a variety of different configurations, depending upon the desired application(s). By way of example and as shown in
Alternatively, individual waveguide array substrates may include multiple originating waveguides, e.g., like originating waveguides 104 and 304 from
The arrays may include parallel waveguides, e.g., as shown, or may include patterned waveguides that have a variety of different configurations, including serpentine waveguides, divergent waveguides, convergent waveguides or any of a variety of configurations depending upon the desired application. For example, where it is desired to provide evanescent illumination to larger areas of the substrate, it may be desirable to provide such serpentine waveguides, wider or slab waveguide(s), or alternatively and likely preferably, larger numbers of parallel or similarly situated waveguides. As noted previously, the waveguide substrates may include a single waveguide that may span a fraction of the width of the substrate or substantially all of that width. In accordance with preferred aspects however, waveguide arrays are used to split individual originating beams into two or more waveguides, preferably more than 10 waveguides, more than 20 waveguides, more than 40 waveguides, and in some cases more than 50 waveguides or even more than 100, 1000, 5000 or more waveguides. The number of waveguides may typically vary greatly depending upon the size of the substrate used, and the optical resolution of the detection system, e.g., its ability to distinguish materials proximal to different waveguides.
The waveguides may individually vary in the size of the core region in order to vary the evanescent field that one can access. Typically, the waveguides will have a cross sectional dimension of from about 0.1 to about 10 μm, preferably from about 0.2 to about 2 μm and more preferably from about 0.3 to about 0.6 μm. A variety of other waveguide dimensions may be employed as well, depending upon the desired application. For example, in some cases, a single waveguide may be used where the cross-sectional dimension of the waveguide is substantially the same as the substrate width or length, e.g., a single waveguide that substantially spans a substrate's width. Notwithstanding the foregoing, preferred aspects will provide arrayed waveguides, e.g., multiple waveguides typically arranged in parallel linear format.
Although described above in terms of a particular type of waveguide structure, e.g., an embedded waveguide structure, a variety of different waveguide structures are exploitable in the present invention, and are shown in
In some cases, the waveguides described herein are generally produced using conventional ion implantation techniques to selectively ion dope selected regions of SiO2 substrates to provide patterned regions of higher refractive index, so as to function as waveguides embedded in the underlying substrate. Examples of such devices are disclosed in, e.g., Marcuse, Theory of Dielectric Optical Waveguides, Second Ed. (Academic Press 1991). Alternate waveguide fabrication processes and configurations are equally applicable to the present invention, including hybrid material waveguides, e.g., employing polymeric materials as a portion or all of the subject substrate, e.g., a polymer core having a first refractive index, disposed within a substrate of another material having a second refractive index, which may be polymeric, or another material, e.g., glass, quartz, etc.
Additional optical functionalities may be provided upon or within the substrates of the invention, including, e.g., providing additional optical confinements upon the substrate, such as zero mode waveguides as discussed in U.S. Pat. Nos. 6,991,726 and 7,013,054, previously incorporated herein by reference. Other optical functionalities that may be integrated into or upon the substrates including, e.g., mask layers, lenses, filters, gratings, antireflective coatings, or the like. Other functionalities may be incorporated into the fabricated substrate that operate on and/or in conjunction with the waveguides or waveguide arrays of the invention. For example, optical switching or attenuation components may be provided upon or within the substrates of the invention to selectively direct and/or modulate the light passing through a given waveguide or waveguides.
By way of example, the waveguide array may have a controllable optical switch or attenuator built into its structure which can provide control over the amount of light allowed to enter the waveguide structure. Such control permits the careful selection of optimal light levels for a given analysis being carried out using the substrate. Further, using independently activated switches or attenuators on a waveguide array permits one to independently control light application to one or a subset of waveguides in an array. In still other advantageous applications, in combination with multiple light sources coupled to the same waveguides through different originating waveguides, optical switching of the input waveguides will permit one the ability to select the light source(s) for any subset or all of the waveguides at a given time, or even to modulate the intensity of a light from a given source, on the fly. By controlling the light from individual sources, e.g., where such sources have differing spectra, one can consequently control the wavelength of the light reaching a given waveguide and its associated reaction regions.
In addition to the ability to more precisely control the light parameters of the overall system for precise tuning of the application, the controlled aspect of the light application provides further abilities to mitigate potential adverse effects of excessive illumination on reaction components, such as photo-damage effects on reactants, or other reaction components resulting from prolonged high intensity illumination.
A variety of different optical devices may be employed in controlling light passage through the waveguides used in the substrates of the invention. In particular, optical modulators, such as Mach-Zhender modulators (see, e.g., U.S. Pat. No. 7,075,695 for discussion of high speed Mach-Zhender modulators), Michelson modulators, thermally tunable modulators that may include other optical devices (see, e.g., Published U.S. Patent Application No. 2005-0232543 for a discussion of thermally tunable modulators) or may employ heating elements to modulate the refractive index of one or more waveguides, optical switches, and the like.
In addition to the optical functionalities of the substrates of the invention, in some cases, such substrates may include additional functionalities that provide a defined region on the substrate surface to limit the access that reagents or other elements have to the illumination zone above a waveguide. For example, in some cases, the substrates may include a patterned structure or set of structures over the surface of the substrate providing selected exposure of the surface exposed waveguide(s). Such selected regions may provide limited areas of illumination on a given substrate by blocking the illumination region existing above other portions of the waveguide(s). As a result, only selected portions of the surface will be within the illumination region of the waveguides. Such regions may be selected to align with detection systems or the requirements of such systems, e.g., sample spacing permitting spectral separation of signals from each region (See, e.g., U.S. patent application Ser. No. 11/704,733, filed Feb. 9, 2007, which is incorporated herein by reference in its entirety for all purposes). In addition to limited access, such structures may also provide structural confinement of reactions or their components, such as wells or channels. In one aspect, for example, microfluidic channels may be provided disposed over surface exposed waveguide or waveguide array. Such channels may be independently used to deliver different reagents to different portions of a waveguide or waveguide array.
Substrates including mask layer 604 may be prepared by a variety of known fabrication techniques. For example, lithographic techniques may be used to define the mask layer out of polymeric materials, such as photoresists, using e.g., conventional photolithography, e-beam lithography, or the like. Alternatively, lithographic techniques may be applied in conjunction with layer deposition methods to deposit metal mask layers, e.g., using aluminum, gold, platinum, chrome, or other conventionally used metals, or other inorganic mask layers, e.g., silica based substrates such as silicon, SiO2, or the like. In particularly preferred aspects, both the underlying substrate and the mask layer are fabricated from the same material, which in particularly preferred aspects, is a transparent substrate material such as an SiO2 based substrate such as glass, quartz, or fused silica. By providing the mask and underlying layers of the same material, one can ensure that the two layers have the same interactivity with the environments to which they are exposed, and thus minimize any hybrid surface interactions.
In the case of SiO2 based substrates and mask layers, conventional fabrication processes may be employed. In particular, a glass substrate bearing the surface exposed waveguide has a layer of resist deposited over its surface. A negative of the mask layer is then defined by appropriate exposure and development of the resist layer to provide resist islands where one wishes to retain access to the underlying waveguide. The mask layer is then deposited over the surface and the remaining resist islands are removed, e.g., through a lift off process, to provide the openings to the underlying waveguides. In the case of metal layers, deposition may be accomplished through a number of means, including evaporation, sputtering or the like. Such processes are described in, e.g., U.S. Pat. No. 7,170,050, which is incorporated herein by reference in its entirety for all purposes. In the case of silica based mask layers, a CVD process may be employed to deposit a silicon layer onto the surface. Following lift off of the resist layer, a thermal oxidation process can convert the mask layer to SiO2. Alternatively, etching methods may be used to etch access points to underlying waveguides using conventional processes. For example, a silicon layer may be deposited over an underlying waveguide substrate. A resist layer is then deposited over the surface of the silicon layer and exposed and developed to define the pattern of the mask. The access points are then etched from the silicon layer using an appropriate differential etch to remove silicon but not the underlying SiO2 substrate. Once the mask layer is defined, the silicon layer is again converted to SiO2 using, e.g., a thermal oxidation process.
In addition to the advantages of reduced auto-fluorescence, waveguide substrates having an integrated mask layer provide advantages of optical alignment over similar arrays of wells or structures. In particular, in illuminating an ordered array of reaction regions with minimal excess illumination, one typically presents a corresponding array of illumination spots. In doing so, one must take substantial care in aligning the optical presentation of the illumination spots to the ordered array of reaction regions. Such alignment presents challenges of both design and robustness, as such systems may be prone to drifting or other misalignment influences. Where, as in the present invention, illumination is hard wired into the substrate by virtue of the integrated waveguide, alignment is automatic.
In other cases, surface features may include other confinement strategies, including, e.g., chemical surface functionalities that are useful in a variety of surface analytical operations, such as hydrophobic coatings or hydrophilic coatings that are optionally patterned, to provide confinement or direction to aqueous materials, chemical derivatization, e.g., to facilitate coupling of other functional groups or otherwise, e.g., by providing hydrophobic barriers partially or completely surrounding a desired region. As will be appreciated, in some cases, particularly where structural confinement is provided upon the surface of the substrate, it may not be necessary to divide up light through a series of discrete waveguides in a given substrate. In particular, because one can obtain a desired level of multiplex and spatial separation organization from structurally dividing up the surface, one need not obtain that property through the use of separate waveguides. In such cases, a single field waveguide disposed at the surface of the substrate will suffice to deliver light to the various reaction regions on the surface of the substrate, e.g., as defined by the mask layer. An example of this is illustrated in
In addition to structures and strategies that provide for positioning and/or confinement upon a substrate surface, other components may be provided upon a substrate, including backside coatings for the substrate, e.g., antireflective coatings, optical indicator components, e.g., structures, marks, etc. for the positioning and or alignment of the substrate, its constituent waveguides, and/or for alignment of other components. Other components may include substrate packaging components, e.g., that provide fluidic interfaces with the substrate surface, such as flow cells, wells or recesses, channel networks, or the like, as macrostructures as compared to the surface defined structures above, as well as alignment structures and casings that provide structural protection for the underlying substrates and interactive functionality between the substrates and instrument systems that work with/analyze the substrates.III. Methods and Applications
A. Generally
As noted previously, the substrates, systems and methods of the invention are broadly applicable to a wide variety of analytical methods. In particular, the substrates of the invention may be employed in the illumination mediated analysis of a range of materials that are disposed upon or proximal to the substrate's surface. Such analyses include, inter alia, a number of highly valued chemical, biochemical and biological analyses, including nucleic acid analysis, protein interaction analysis, cellular biology analysis, and the like.
B. Exemplary Applications
1. Sequencing by Synthesis
One example of an analytical operation in which the present invention is particularly applicable is in the determination of nucleic acid sequence information using sequence by synthesis processes. Briefly, sequencing by synthesis exploits the template directed synthesis of nascent DNA strands, e.g., using polymerase mediated strand extension, and monitors the addition of individual bases to that nascent strand. By identifying each added base, one can deduce the complementary sequence that is the sequence of the template nucleic acid strand. A number of "sequence by synthesis" strategies have been described, including pyrosequencing methods that detect the evolution of pyrophosphate upon the incorporation of a given base into the nascent strand using a luminescent luciferase enzyme system as the indicating event. Because the indicator system is generic for all four bases, the process requires that the polymerase/template/primer complex be interrogated with only one base at a time.
Other reported sequence by synthesis methods employ uniquely labeled nucleotides or nucleotide analogs that provide both an indication of incorporation of a base, as well as indicate the identity of the base (See, e.g., U.S. Pat. No. 6,787,308). Briefly, these methods employ a similar template/primer/polymerase complex, typically immobilized upon a solid support, such as a planar or other substrate, and interrogate it with nucleotides or nucleotide analogs that may include all four bases, but where each type of base bears an optically detectable label that is distinguishable from the other bases. These systems employ terminator bases, e.g., bases that, upon incorporation, prevent further strand extension by the polymerase. Once the complex is interrogated with a base or mixture of bases, the complex is washed to remove any non-incorporated bases. The washed extended complex is then analyzed using, e.g., four color fluorescent detection systems, to identify which base was incorporated in the process. Following additional processing to remove the terminating group, e.g., using photochemistry, and in many cases, the detectable label, the process is repeated to identify the next base in the sequence. In some cases, the immobilized complex is provided upon the surface as a group of substantially identical complexes, e.g., having the same primer and template sequence, such that the template mediated extension results in extension of a large number of identical molecules in a substantially identical fashion, on a step wise basis. In other strategies, complexes are immobilized in a way that allows observation of individual complexes resulting in a monitoring of the activity of individual polymerases against individual templates.
As will be appreciated, immobilization or deposition of the polymerase/template/primer complex upon or proximal to the surface of the waveguide core in the waveguide arrays of the invention will allow illumination, and more notably in the case of fluorescence based assays, excitation, at or near selected regions of the surface without excessive activation and fluorescence interference from the surrounding environment, which can be a source of significant noise in fluorescence based systems.
In another sequencing-by-synthesis process, one monitors the stepwise addition of differently labeled nucleotides as they are added to the nascent strand and without the use of terminator chemistries. Further, rather than through a one-base-at-a-time addition strategy, monitoring of the incorporation of bases is done in real time, e.g., without the need for any intervening wash steps, deprotection steps or separate de-labeling steps. Such processes typically rely upon optical strategies that illuminate and detect fluorescence from confined reaction volumes, such that individual complexes are observed without excessive interference from labeled bases in solution that are not being incorporated (See U.S. Pat. Nos. 6,991,726 and 7,013,054, previously incorporated herein, and 7,052,847, 7,033,764, 7,056,661, and 7,056,676, the full disclosures of which are incorporated herein by reference in its entirety for all purposes), or upon labeling strategies that provide fluorescent signals that are indicative of the actual incorporation event, using, e.g., FRET dye pair members on a base and on a polymerase or template/primer (See U.S. Pat. Nos. 7,052,847, 7,033,764, 7,056,661, and 7,056,676, supra).
This aspect of the invention is schematically illustrated in
In accordance with the foregoing sequence by synthesis methods, one may optionally provide the complexes over an entire surface of a substrate, or one may selectively pattern the immobilized complexes upon or proximal to the waveguide cores. Patterning of complexes may be accomplished in a number of ways using selectively patternable chemical linking groups, and/or selective removal or ablation of complexes not in the desired regions. In some cases, one can employ the waveguides in selectively patterning such complexes using photoactivatable chemistries within the illumination region. Such strategies are described in detail in U.S. patent application Ser. No. 11/394,352 filed Mar. 30, 2006, and incorporated herein by reference in its entirety for all purposes.
In addition to selective immobilization, and as noted above, in some cases it is desirable to immobilize the complexes such that individual complexes may be optically resolvable, e.g., distinguished from other complexes. In such cases, the complexes may be immobilized using highly dilute solutions, e.g., having low concentrations of the portion of the complex that is to be immobilized, e.g., the template sequence, the polymerase or the primer. Alternatively, the surface activation for coupling of the complex component(s) may be carried out to provide a low density active surface to which the complex will be bound. Such surfaces have been described in U.S. patent application Ser. No. 11/240,662, filed Sep. 30, 2005, which is incorporated herein by reference in its entirety for all purposes. Again, such low density complexes may be patterned just upon or proximal to the waveguides or they may be provided across the surface of the substrate, as only those reaction complexes that are proximal to the waveguides will yield fluorescent signals.
While described in terms of real-time nucleic acid sequencing by synthesis, it will be appreciated that a wide variety of real-time, fluorescence based assays may be enhanced using the substrates, methods and systems of the invention.
2. Molecular Arrays and Other Surface Associated Assays
Another exemplary application of the substrates and systems of the invention is in molecular array systems. Such array systems typically employ a number of immobilized binding agents that are each specific for a different binding partner. The different binding agents are immobilized in different known or readily determinable locations on a substrate. When a fluorescently labeled material is challenged against the array, the location to which the fluorescently labeled material binds is indicative of it's identity. This may be used in protein-protein interactions, e.g., antibody/antigen, receptor-ligand interactions, chemical interactions, or more commonly in nucleic acid hybridization interactions. See, U.S. Pat. Nos. 5,143,854, 5,405,783 and related patents, and GeneChip® systems from Affymetrix, Inc.
In accordance with the application of the invention to arrays, a number of binding regions, e.g., populated by known groups of nucleic acid probes, are provided upon a substrate surface upon or proximal to the waveguides such that a hybridized fluorescently labeled probe will fall within the illumination region of the waveguide. By providing for selective illumination at or near the surface, one can analyze hybridized probes without excessive interference from unbound fluorescent materials.
This aspect of the invention is schematically illustrated in
3. Cellular Observation and Analysis
In still another exemplary application, cell based assays or analyses may be carried out by providing cells adhered to the substrate surface over the waveguides. As a result, one could directly monitor fluorescently labeled biological functions, e.g., the uptake of fluorescent components, the generation of fluorescent products from fluorogenic substrates, the binding of fluorescent materials to cell components, e.g., surface or other membrane coupled receptors, or the like.
4. Other Applications
It will be appreciated by those of ordinary skill that the substrates of the invention may be broadly applicable in a wider variety of applications that analytical processes. For example, such substrates and methods may be employed in the identification of location of materials on surfaces, the interrogation of quality of a given process provided upon the surface, the photo-manipulation of surface bound materials, e.g., photo-activation, photo-conversion and/or photo-ablation. As such, while some of the most preferred applications of the present invention relate to analytical operations and particularly in the fields of chemistry, biochemistry, molecular biology and biology, the discussion of such applications in no way limits the broad applicability of the invention.IV. Systems
In general, the substrates of the invention are employed in the context of other components as a system or one or more subsystems. By way of example, in preferred aspects, the substrates of the invention are employed in the analysis of materials disposed upon the substrate surface as described elsewhere herein. In such cases, the substrates of the invention are generally exploited in conjunction with and as a part of an analytical substrate and reagent system that is used in the desired analysis. Such reagent systems may include proteins, such as enzymes and antibodies, nucleic acids including nucleotides, nucleosides, oligonucleotides and larger polymers of same, substrates for a given reaction, cells, viruses or phages, or any of a variety of different chemical, biochemical or biological components for a desired analysis.
In addition to the "wet-ware" components of the systems set forth initially above, the invention also includes the substrates of the invention in conjunction with hardware and/or software system components. As noted previously, such hardware components include, e.g., optical components such as lenses, mirrors, prisms, illumination sources and the like, detection systems, while software components include processes for controlling overall systems and/or software for processing, evaluating and presenting data derived from those systems.
A. Reagent Systems and Kits
As set forth above, the substrates of the invention may be applied and/or packaged in kits with other reagents, buffers and other adjuncts that are used in the desired analysis. The nature of such reagents is generally application specific and will vary according to such applications. By way of example in application of such substrates to nucleic acid sequencing methods, as described below, the substrates may include one or more of template nucleic acids, nucleic acid probes, polymerase enzymes, native and/or normative nucleotides or nucleotide analogs, that will in certain preferred aspects, include labeling groups such as fluorescent labels. One or more of the foregoing components may be either packaged with and/or applied as immobilized components on the surface of the substrate that is exposed to the evanescent radiation from the waveguides.
In the context of application in real time sequencing by synthesis, reagent systems may include polymerase enzymes, e.g., as described in U.S. Patent Application No. 60/753,515 filed Dec. 22, 2005, which is incorporated herein by reference, or commercially available polymerase enzymes, e.g., taq polymerases, Sequanase® polymerases, Phi 29 polymerases, DNA Polymerase I, Klenow fragment, or any of a variety of other useful polymerases known to those of skill in the art. Additionally, such systems may include primer sequences that are either specific for a particular template sequence or are specific for a universal tag sequence that may be ligated to or otherwise provided in line with the template sequence. Such systems may further include nucleotides or nucleotide analogs, such as fluorescently labeled nucleotides like those described in U.S. patent application Ser. Nos. 11/241,809, filed Sep. 29, 2005, Published U.S. Application No. 2003/0124576, and U.S. Pat. No. 6,399,335, the full disclosures of which are all incorporated herein by reference for all purposes.
B. Optics and Instrumentation
As noted elsewhere herein, instrument systems are also included for use in conjunction with the substrates and methods of the invention. Typically, such systems include the substrates of the invention interfaced with appropriate light sources, such as one or more lasers, for delivering desired electromagnetic radiation through the waveguides. Also included is an appropriate optical train for collecting signals emanating from the substrate surface following illumination, and detection and data processing components for detecting, storing and presenting signal information.
One example of a system for use in the present invention is illustrated in
While illustrated with a first light source, e.g., laser 904, and an optional second light source, e.g., optional laser 906, it will be appreciated that additional light sources may be provided optically coupled to the waveguide arrays, e.g., using additional originating waveguides to direct light from each the various sources to all or a subset of the waveguides in a given array. For example, in some cases, 3 light sources, 4 light sources or more may be used. Additional light sources will preferably provide light having different spectral characteristics, e.g., peak wavelengths, to the waveguides, although they may also be employed to provide additional intensity or variations in other light characteristics, such as frequency.
The detection system is typically configured to detect signals from large areas of the substrate, e.g., multiple signals emanating from a plurality of different regions on the substrate, and preferably, do so simultaneously. Thus while scanning detection optics may be employed for certain applications of the invention, in general, larger area imaging detection systems are preferred.
Other optical trains and detection systems that may be employed in the present invention are described in U.S. patent application Ser. Nos. 11/201,768 (filed Aug. 11, 2005) and 60/772,908 (filed Feb. 13, 2006), each of which is incorporated herein by reference, which, with provision of a waveguide substrate of the invention and direction of excitation illumination through the waveguides, would be directly useful in the context of the invention. In particular, in certain aspects, the imaged signal will be a series of discrete signal sources or points of signal origin on the overall surface of the substrate. As such, the detection systems described in the aforementioned application would be directly applicable to the present invention.
Although described in some detail for purposes of illustration, it will be readily appreciated that a number of variations known or appreciated by those of skill in the art may be practiced within the scope of present invention. To the extent not already expressly incorporated herein, all published references and patent documents referred to in this disclosure are incorporated herein by reference in their entirety for all purposes.
1. An analytical device, comprising:
- a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface; and
- an analyte disposed sufficiently proximal to the first surface and external to the waveguide, to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide.
2. The device of claim 1, wherein the substrate comprises more than one waveguide disposed upon the first surface.
3. The device of claim 1, wherein the at least first waveguide is disposed within the first surface of the substrate.
4. The device of claim 1, wherein the substrate comprises at least 10 waveguides disposed upon the first surface.
5. The device of claim 1, wherein the substrate comprises at least 20 waveguides disposed upon the first surface.
6. The device of claim 1, wherein the substrate comprises at least 40 waveguides disposed upon the first surface.
7. The device of claim 2, wherein a plurality of the more than one waveguides disposed upon the first surface are optically coupled to a common originating waveguide.
8. The device of claim 1, wherein the substrate comprises SiO2.
9. The device of claim 1, wherein the substrate comprises a polymer.
10. The device of claim 1, further comprising at least a first excitation radiation source optically coupled to the waveguide to pass excitation light therethrough.
11. The device of claim 10, wherein the at least first excitation radiation source is coupled to the first optical waveguide by at least a first optical fiber.
12. The device of claim 1, further comprising at least two excitation radiation sources optically coupled to the first optical waveguide to pass excitation light therethrough, each of the two excitation radiation sources providing light of a different spectrum.
13. The device of claim 1, further comprising a mask layer disposed over the first surface of the substrate, the mask layer covering at least a first portion of the waveguide on the first surface and not covering at least a second portion of the waveguide on the first surface.
14. The device of claim 13, wherein the mask layer comprises a plurality of apertures disposed therethrough, the apertures providing access to the at least a portion of the waveguide on the first surface.
15. The device of claim 13, wherein the mask layer and the substrate layer comprise the same material.
16. The device of claim 15, wherein the same material comprises a silica based material.
17. The device of claim 16, wherein the silica based material comprises SiO2.
18. The device of claim 13, wherein the mask layer comprises a polymer.
19. The device of claim 13, wherein the mask layer comprises a metal.
20. A method of illuminating an analyte, comprising:
- providing a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface;
- providing an analyte disposed sufficiently proximal to the first surface and external to the waveguide, to be illuminated by an evanescent field emanating from the waveguide when light is passed through the waveguide; and
- directing light through the first waveguide such that the evanescent field from the waveguide illuminates the analyte.
21. The method of claim 20, wherein the analyte is confined proximal to a selected portion of the first surface.
22. The method of claim 21, wherein a structural barrier confines the analyte proximal to the selected portion of the first surface.
23. The method of claim 24, wherein the structural barrier comprises a well in a mask layer disposed over the first surface, the well providing confinement of the analyte proximal to a portion of the first surface.
24. The method of claim 20, wherein the analyte is immobilized proximal to the first surface.
25. The method of claim 20, wherein the analyte comprises a fluorescent moiety, and the step of directing light comprises directing light having an excitation wavelength for the fluorescent moiety.
26. The method of claim 20, wherein the analyte is confined proximal to a portion of the first surface by a hydrophobic region of the first surface.
27. The method of claim 26, wherein the hydrophobic region surrounds the portion of the first surface.
28. The method of claim 20, wherein the substrate comprises at least a first fluidic channel disposed over and in fluid communication with the first surface, and the providing step comprises delivering the analyte proximal to the first surface through the first fluidic channel.
29. The method of claim 20, wherein the analyte comprises a complex of a polymerase enzyme, a target nucleic acid sequence, a primer sequence complementary to at least a portion of the target sequence, and a nucleotide or nucleotide analog.
30. A system for analyzing an analyte, comprising:
- a substrate comprising a first surface and at least a first optical waveguide disposed upon the first surface;
- at least a first light source optically coupled to the at least first waveguide to direct light into the first waveguide; and
- an optical detection system positioned to receive and detect optical signals from an analyte disposed sufficiently proximal to the first surface and external to the waveguide to be illuminated by an evanescent field emanating from the waveguide when light from the light source is passed through the waveguide.
31. The system of claim 11, wherein the optical detection system receives and detects signals from a plurality of locations on the surface of the substrate.
32. The system of claim 30, wherein the analyte comprises a fluorescent moiety that is excited by light of the wavelength provided by the first light source, and wherein the optical detection system comprises a detector for detecting fluorescent signals, and an optical train positioned to receive fluorescent signals from the first surface of the substrate and direct the fluorescent signals to the detector.
33. The system of claim 30, further comprising at least a second light source optically coupled to the at least first waveguide.
34. The system of claim 30, wherein the substrate comprises a plurality of optical waveguides disposed upon the first surface of the substrate.
35. The system of claim 30, wherein the substrate comprises at least 10 waveguides disposed upon the first surface.
36. The system of claim 30, wherein the substrate comprises at least 20 waveguides disposed upon the first surface.
37. The system of claim 30, wherein the substrate comprises at least 40 waveguides disposed upon the first surface.
Type: Application
Filed: Oct 31, 2007
Publication Date: Jun 26, 2008
Patent Grant number: 7834329
Applicant: Pacific Biosciences of California, Inc. (Menlo Park, CA)
Inventors: Paul LUNDQUIST (San Jose, CA), Stephen TURNER (Menlo Park, CA)
Application Number: 11/981,795
International Classification: G02B 6/26 (20060101);


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This giant slab of titanium delight certainly has all the aesthetics of a top-notch water cooling sytem.
The external tank has three LED dials and the coloured coolant snakes its way into your case with the tech-cool credentials of a Japanese tailor.
The Reserator's set-up is relatively simple too - with patience and a pair of pliers the risk of leakage is minimal.
Lacklustre cooling
Where we have doubts, though, is the straightforward air cooling of the fluid. Essentially, it means that the actual cooling capability of the Zalman isn't as good as a £30 air fan, and it's not totally silent either.
Still, if you rig up the GPU block too, you can do a fair job of quieting a noisey games PC, but for high-end overclocking you'll probably need to invest in some expensive phase changing goodness. It's available from QuietPC.
Test results:
Full resource draw stress test: Prime 95: 46°C
Max load multi-core stress test: HL2 map draw: 55°C
Real-world gaming stress test: World in Conflict: 48°C
Idle recovery (1 min after stress test): Resting: 37°C


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Find a new car near Bristol, RI on YaSabe. Use our website to find new cars and get new car prices. You can also choose from any of the following links to help in your search for a new car.
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The Super Bowl of Food: Pittsburgh vs. Green Bay
It's Super Bowl time, and at Serious Eats, that means the same showdown we do every year: which team wins for the best food city?
This year, we'll give each city a wide berth—say, 150 miles, or until you encroach on another team's territory. After all, Packers fans don't just live in Green Bay.
So if delicious foods were predicting the Super Bowl score, in our necessarily incomplete and arbitrary judging, how would the game play out? Follow along with us, quarter by quarter.
First Quarter: Burgers and Sandwiches
The first quarter is a battle of food between bread, and it's tough competition indeed. In Green Bay territory, you've got crazy butter burgers and perhaps my favorite new chef burger of the year, at Graze in Madison; but Pittsburgh has one of our man Nick Solares's very favorite burgers at Tessaro's.
Chain burger territory? It's Culver's v. Steak 'n Shake, but we've got to give the nod to the latter.
Sandwiches? Wisconsin doesn't have an iconic sandwich the way that Pittsburgh has Primanti's, but that's not to say there aren't sandwiches worth eating; one of the surprises of my travels this year was the corned beef from Jake's in Milwaukee. Bite for bite, I'd prefer Jake's, but history matters, too.
Score? 7 Packers, 6 Steelers. Both scored major victories, but Pittsburgh just didn't get that last point.
Second Quarter: Food Legacies
Which region has made the bigger mark on the food world? Pittsburgh, of course, is the home of Heinz—and that's a national and international legacy that's hard to beat. Latrobe, not far outside Pittsburgh, is the home of Rolling Rock; and Klondike bars weren't invented far away, just across the state line in Youngstown (which, our in-house Steelers expert assures us, is firmly Steeler country).
But of course, any beer drinker knows Wisconsin as the original home of Miller, Pabst, and many more—not to mention cheeses, and brats and sausages including Usinger's. And what about present-day food culture? Pittsburgh's got the still-active Strip District, a happy place for food shoppers, but Madison's got one of the most impressive farmers' markets we know.
Another tough one. Of course the mega-breweries score Green Bay points, but beer plus ice cream plus the one bottle on every diner table in America gets Pittsburgh more.
Score? 7 Steelers, 3 Packers.
Third Quarter: Pizza and Fried Foods
Ah, my favorite quarter. Pittsburgh has a somewhat surprising pizza legacy (Il Pizzaiolo was a VPN-certified Neapolitan pizzeria years before there was a 1000° oven in every city), but Wisconsin's got its own idiosyncratic joints, our favorite of which is Maria's in Milwaukee. In Sheboygan, there's also Il Ritrovo. The pizza round is a wash—a touchdown each.
Fried foods? Pittsburgh is a proud pierogi town, no doubt about it, and it's not as if they can't get their fry on—heck, they put French fries on their most famous sandwiches, which has to count for something. But between fried cheese curds and fish fries, Wisconsin is the decisive victor—a touchdown to Pittsburgh's field goal.
Score? 14 Packers, 10 Steelers. Huge quarter.
Fourth Quarter: Sweets and Desserts
It'd be unfair not to recognize the deliciousness of the hotcakes from Pamela's in Pittsburgh, so breakfast sweets count too. But Wisconsin's got some up-and-coming bakeries, like the charming Batch Bakehouse in Madison.
As far as desserts go? Pittsburgh may have tasty ice cream and sweets, and old-school bakeries like Prantl's, but frozen custard all over Wisconsin is absolutely unequaled (Leon's, Kopp's, North Point, Michael's). And that's just the start of it.
Score? 7 Packers, 3 Steelers.
Final Score
It was a nail-biter to the end, but the final score is 31 - 26 in favor of the Packers—though, to be fair, with a lot of help from its supporting cities, whereas Pittsburgh gets almost none.
What say you, eaters? What's your prediction for Sunday's game?
All products linked here have been independently selected by our editors. We may earn a commission on purchases, as described in our affiliate policy.


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- Calendar
- Food and Drink
- Date
- 22nd September 5:45pm
- Location
We offer a limited number of Pre Show meal tickets at a special discounted rate of £14.50. These tickets include 2 courses from a pre-theatre set menu – don't forget to book your show tickets too.
How Does it Work?
When you book your Pre Show Meal tickets (either online or over the phone), your table is automatically reserved in the café bar. You must take your table by 90 minutes before the performance. We will reserve your table in the café bar under the same name that the tickets were booked in. Please note drinks and service are not included in the Meal Deal.
How Much Does It Cost?
The price of a two course meal is £14.50. The on the day price will vary as the menu changes.
What Will Be On The Menu?
The menu changes regularly and will always include a vegetarian option. You can check out the current menu by clicking the link below.
Location information
New Wolsey Theatre
- Country
- United Kingdom
- County
- England
- City
- Ipswich IP1 2AS


text:
The Sky Is Falling
The sky is falling, actually.
Well, sort of.
We had a scary incident this week at Car Talk Plaza in Hahvahd Squayah. Fortunately, no one was hurt.
What happened? The old concrete ceiling in our Producer Dougie Berman's office actually came crashing down. Among the rubble was a chunk of ceiling that landed right on Dougie's chair. Had he been napping there at the time, with his head tilted back... it would have rammed itself right down his gullet - and Dougie would be history.
From this little workplace safety incident, Tom and Ray drew an important lesson:
Beware of work.
Had Dougie been more diligent about being at work, he might not be here right now (asleep in his temporary office next door). So, let this be a lesson to us all:
Don't go to work. It's dangerous!


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- This event has passed.
South Devon Coffee Afternoon coffee, cake and catch up
12th November 2018 @ 1:30 pm - 3:00 pm
We would like to invite you along to our coffee, cake and catch-up afternoons. The aim of this is for local adoptive families to come together for peer support. Children under 5 can explore and play with the toys ; however all parents are more than welcome if your children are at school. You can come along and enjoy a coffee from the cafe with a complementary piece of cake for our families.
Please contact us for venue details


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Aircraft Rescue Firefighters (ARFF)
If you get sick at the airport, you'll likely have several Emergency Medical Technicians (EMTs) at your side within two minutes—the airport has a full time fire department and every firefighter has Firefighter 1 or Firefighter 2 certification and is a certified EMT. Besides responding to routine medical calls, ARFF members stand ready to respond to the worst case scenario—a crashed or burning aircraft.
Airport Police
The airport community is protected by the Airport Police Department (APD). With an authorized strength of 10 full time commissioned officers, the department provides the airport with 24/7 police protection. Airport policing involves extensive customer service and officers frequently find themselves assisting customers to ensure their visit is a pleasant experience.
Protecting the public and providing exceptional customer service are the main priorities of the APD. APD officers have at least two years prior police experience before being hired. Officers at the airport have a thorough knowledge of airport security issues and police methods applicable to the airport environment.
The APD achieved status as a criminal justice agency in June of 2002 and it is recognized by Missouri Department of Public Safety as a stand alone law enforcement agency.
Contact the APD at 417.868.0580.
Public Safety Resources
Irregular Operations (IROPS) Contingency Plan


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