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Kusudama Cloche
If you were visiting during the Fall/Christmas season last year, you might remember the Kusudama Balls that I made from old book pages. I had a LOT of time in waiting rooms for doctors and surgeries last year and I kept supplies in my purse and made tons of the little pieces to pass the time. I ended up with some awesome decorations! There were three posts here, here and here. These are some of those pictures in case you don't want to go look.
They were so pretty that I didn't put them away after the holiday, but I've been concerned about dust. How do you dust these? They took so long to make and I want them to stay nice for a long time. I had an idea.
I fell in love with cloches in Pottery Barn catalogs, but wow are they out of the realm of possibility! So, like any good DIY'er I kept my eyes open for more affordable things that would give the same look. I bought a cloche and base from Hobby Lobby for $13.50 on half price sale.
I used this cloche in my Pottery Barn knock off spring tablescape here.
Ever since taking the spring table down so I could use the table for The Accidental Remodel of our kitchen, I had planned to use the cloche for the Kusudama Balls. There was so much to do in the remodel and the house was a wreck from having the entire kitchen moved into all the other rooms. I only got to it working on the cloche recently.
The redish brown base worked fine on the spring tablescape, but not for the Kusudama Balls. I used the same primer and paint as my cabinets and trim in the kitchen on the base. I had painted the trim in the rest of the house and our coffee table with the same paint before I started the kitchen. So, anywhere in the public part of the house, this lovely soft white base will go perfectly! And if I use it in the kitchen some time, NOTHING looks as good on my black granite counters as a soft white!
Do you remember by Pussy Willow Ball? You can read about it here. It too has been a dust concern and I do love it so. The colors are awesome and it's soooooooo soft!
I finally gathered all my dust endangered treasures from last fall together in the cloche! I love it and no more dust issues! The pussy willow ball is on a spindle of twine. The six pointed Kusudama ball is on a Bath & Body Works candle in it's silver metal snowflake holder. The five pointed Kusudama ball is sprinkled with "diamonds" from Hobby Lobby. They are sold to sprinkle on your table. I got them 50% off for about $1.50.
I may weather the base with sandpaper in the future, but for now it's on a coffee table that isn't weathered and it looks just fine. My husband says it looks snowy. Practical, affordable and beautiful! Yeah! Chalk another one up for DIY!
I'm posting this to The Scoop at Confessions of a Plate Addict and The Sunday Showcase.
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This is my contribution to the BBD# 33: Breads with booze which I have the pleasure to host this month. There's still plenty of time to participate, so go for it!
For the sweet dough
From 'Artisan Baking'
Makes 18 rolls
½ cup milk
2 teaspoons instant yeast
2 2/3 cup unbleached all-purpose flour
3 eggs
1 ½ teaspoons table salt
6 tablespoons granulated sugar
½ cup unsalted butter
Microwave the milk for 4 min or heat it in a pan until bubbles form around the edges and the milk smells cooked. Let it cool to 105 F and dissolve the yeast in the milk. (Scalding the milk denatures the proteins that prevent gluten formation). Let it stand for 10 minutes.
In a large bowl, add the flour and then the yeast mixture and eggs. With a wooden spoon or your hand, mix until combined. Cover the bowl with plastic and let it stand for 15 to 20 min.
Add the salt to the dough and mix until combined. Take the dough out of the bowl and knead until smooth, for about 10 min. The salt will incorporate just fine. Add half the sugar and keep kneading, then add the other half. Finally, add the butter in 2 additions and knead it into the dough until this is satiny and soft.
Place the dough in a covered container in the fridge, for at least 8 hrs and up to 48.
Makes 9 rolls
4 very ripe bananas
½ cup bourbon
1 teaspoon cinnamon
Cut the bananas into big chunks and cook in a pan with the bourbon until all the liquid is absorbed. Transfer the bananas to a bowl and mash together with the cinnamon. Let the filling cool.
To assemble the rolls
Roll the sweet dough into a rectangle, ½ inch in thickness. Spread the cooled banana filing and roll the dough like you would do with a swiss roll. Cut the dough into 1 to 1 ½ inch pieces and place them in a round pan. Let them rise for 30 min. In the meantime, preheat the oven to 325 F. Brush the rolls with egg wash. Bake them for 30-35 minutes or until golden brown on top. Let them cool slightly before eating.

subset: commoncrawl


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On February 4, 2013, Dr. William R. Jacobs, Jr., was awarded the seventh annual Marshall S. Horwitz, M.D. Faculty Prize for Research Excellence. Dr. Jacobs delivered his prize lecture, "Unmasking the Houdini Traits of Mycobacterium tuberculosis Persistence in the Presence of Drugs and Immune Effectors," to an enthusiastic audience in Robbins Auditorium.
Tuberculosis is caused by the bacterium Mycobacterium tuberculosis (MTB). It ranks second only to HIV/AIDS as the greatest worldwide killer caused by as single infectious agent. According to the World Health Organization, 1.4 million people died from tuberculosis in 2011.
MTB infects people through inhalation; it can persist as a latent infection for decades, causing serious illness or death when MTB starts to multiply and the infection reactivates. Dr. Jacobs aptly compared MTB to the master escape artist Harry Houdini. Despite the availability of several drugs that can kill it, MTB has managed death-defying acts that complicate efforts to treat those infected. Its notorious ability to mutate into drug-resistant strains poses a major global health problem.
Considering its deceptive nature, it's appropriate that the scientist dubbed the "TB Terminator" by the American Association for the Advancement of Science (AAAS) is tackling MTB. In his talk, Dr. Jacobs described his team's strategy in confronting MTB—particularly those MTB strains that are proving resistant to most of the standard anti-TB drugs. Working in a lab that overlooks a former TB sanatorium, he and his colleagues are developing vaccines to prevent disease, diagnostic tools to detect it, and drugs to treat infection.
As part of those efforts, his team is systematically knocking out every MTB gene to find genes that are vital for MTB's survival and therefore might be good targets for drugs aimed at killing the microbe. His lab's diagnostic work focuses on rapidly detecting small numbers of MTB in clinical sputum samples and pinpointing the drugs to which they're susceptible.
The Horwitz Prize was established at Einstein in memory of beloved Einstein faculty and renowned scientist Dr. Marshall S. Horwitz, who passed away in 2005.
Posted on: Wednesday, March 13, 2013
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Assay tray
A machine for transferring liquids to and from the wells of assay trays in a controlled, automated manner and a solid phase assay tray for use with the machine. The machine includes a horizontally translatable table (15) that holds the tray (46), a plurality of liquid dispensing manifolds (54) for dispensing liquids into the tray wells (50) and an aspirating manifold (65) for aspirating liquid from the well. The dispensing and aspirating manifolds are mounted on a vertically translatable head (16) above the table. Each dispensing manifold is equipped with a row of dispensing tubes (56) and is connected via a pump (58) to a liquid container (62). The aspirating manifold is equipped with a row of aspirating tube (66) and is connected via a pump (69) to a waste liquid receptacle (73). A microprocessor (85) controls the movements of the table and manifolds and operates the pumps. The tray wells include means, such as sloping bottoms (74) or subwells (83), that cause the solid phase (76) to occupy a particular position in the wells and sumps (75), (84) that are positioned relative to the location of the solid phase such that they may be accessed vertically by the aspirating tubes without danger of disturbing the solid phase. The bottoms of the sumps have optically flat areas so that beams of light may be passed vertically through the liquid contents of the wells without intersecting the solid phase to make optical measurements of the liquid contents.
Latest Cetus Corporation Patents:
1. Technical Field
This invention is in the field of immunoassay apparatus. More particularly it concerns (a) an immunoassay tray for carrying out an immunoassay in which an immunochemical is fixed to a solid phase and the immunochemical-bearing solid phase is incubated with liquid reagents and washed with wash liquids and (b) a machine for use in combination with the tray for depositing and withdrawing the reagent/wash liquids from the tray wells.
2. Background Art
Immunoassays are used to detect the presence or quantity of a given immunochemical in a sample suspected of containing the immunochemical. Most immunoassays involve the formation of immune complexes via antigen-antibody binding. The complexes are detected in various ways, such as by labels bound directly or indirectly to the complex.
Many immunoassays require that the complexes be subjected to one or more isolations, washings, and treatments with liquid reagents. In order to facilitate handling the immune complexes, one of the components of the complex is used in a solid phase form (i.e., immobilized on a particulate immunosorbent material) which results in the complexes also being in solid phase. The immobilized complexes may be easily washed, incubated with additional reagents, and isolated as required by the particular type of assay. Assays that employ a particulate solid phase reagent are commonly referred to as "solid phase", "heterogeneous", or "immunosorbent" assays. For convenience they will be referred to herein as "solid phase" immunoassays.
Solid phase assays are frequently carried out in assay trays or tubes. Assay trays are plates that have a plurality of wells (usually 20, 24, 48 or 96) arranged in rows and columns into which the particulate solid phase is placed and treated sequentially with the liquid reagents and washes involved in the particular assay. Various well configurations have been used or suggested, including wells having flat bottoms, V-shaped bottoms, and U-shaped bottoms. The liquid reagents and washes are normally added and withdrawn from the wells with manual, semiautomated or automated pipettes.
With most of the current assay trays the location of the solid phase in the well bottom is happenstance. Accordingly, when the pipette tip is inserted into the well there is a possibility of hitting the solid phase and mechanically dislodging bound materials from its surface. Also, the solid phase may interfere with withdrawing liquids completely from the well. In view of this, many assay protocols require that the solid phase be removed from the wells prior to liquid transfer and replaced in the wells after liquid transfer. Also in some types of assays, such as fluorescent immunoassays (FIAs) and enzyme immunoassays (EIAs), the reading step of the assay involves passing a beam of light through the liquid phase. If the solid phase is positioned randomly in the wells it may interrupt the path of the beam, requiring adjustment of the beam path or removal of the solid phase from the well.
Manual, semiautomated or automated pipettes are commonly used for the liquid transfer steps of immunoassays. An example of a fully automated pipette system that may be used to carry out an immunoassay is described in commonly owned U.S. application Ser. No. 06/489,866, filed 5 May 1983. The extensive number of steps and reagents involved in many immunoassays may, however, make it impractical to use such systems where the reagents are carried in open receptacles on a moving table and are transferred via pipette. Also, these pipette systems are generally not capable of carrying out the reading phase of an immunoassay. The investigator must, therefore, transfer the assay tray to reading apparatus. In sum, current assay equipment requires substantial human intervention and is not capable of conducting assays from start-to-finish automatically.
Among the objects of the present invention are to provide a novel assay tray whose wells include means for positioning the solid phase such that it does not interfere with the implement(s) used to introduce and withdraw liquid from the wells or optical measurements made on the liquid contents of the well at the end of the assay and a machine that introduces and withdraws liquids from such wells in an automatic, controlled fashion without necessarily using pipettes and can be adapted to carry out assays automatically from start-to-finish.
Use of the trays of the invention avoids handling of the solid phase, improves the reliability of the assay, and permits solid phase assays to be conducted automatically. The invention machine is particularly adapted to be used in combination with the trays and provides a means for carrying out solid phase assays from start-to-finish with no or only minimal operator monitoring and intervention. In this regard the machine may be readily equipped to conduct the reading phase of the assay automatically and to communicate with a microprocessor to record assay data, correlate such data with other data (if necessary or desired), process or analyze the data, and prepare reports based on the data. When equipped with assay reading means and linked to a microprocessor programmed with suitable software, the present invention provides a self-contained system for conducting, recording, and reporting assays.
There are two principal aspects of the invention: (1) novel assay trays for use in solid phase assays and (2) a machine for supplying and withdrawing liquid reagents to and from the wells of assay trays in an automated controlled manner. These aspects are broadly described below.
The assay trays are used in assays wherein a a particulate solid phase, typically carrying an immobilized reagent, is contacted with one or more liquids. They include means for positioning the solid phase at a predetermined location in the wells combined with other novel structural features that permit liquid to be aspirated from the wells without disturbing the solid phase or the solid phase to be immersed in a minimal volume of liquid or facilitate automated reading of the liquid in the wells.
Thus, in one embodiment the invention tray comprises a plate having a plurality of wells for receiving the solid phase and liquids, the wells having:
(a) a solid phase positioning means for positioning the solid phase in a predetermined location in the wells; and
(b) a sump means spaced from the location of the solid phase whereby liquid may be withdrawn from the wells without disturbing the solid phase.
In another embodiment the invention tray comprises a plate having a plurality of wells for receiving the solid phase and the liquids, the wells each having:
(a) a subwell section for holding the solid phase, the size and shape of the subwell section being such that the solid phase may be immersed in the subwell section in a small volume of liquid; and
(b) a main well section of substantially greater cross sectional area and volume than the subwell section.
The machine comprises in combination:
(a) a head assembly translatable between upper and lower positions along a vertical axis and including:
(i) at least one liquid reagent dispensing manifold having an inlet adapted to be connected to one or more liquid reagent sources and a plurality of spaced outlets each fitted with a vertically depending liquid reagent dispensing tube, the spacing between the dispensing tubes being in correspondence with the spacing between the wells of the assay tray; and
(ii) at least one liquid reagent aspirating manifold having an outlet and a plurality of spaced inlets each fitted with a vertically depending liquid reagent aspirating tube, the spacing between the aspirating tubes being in correspondence with the spacing between the wells of the assay tray;
(b) dispensing pump means operably associated with the source(s) of liquid reagent and the dispensing manifold inlet(s) for pumping liquid reagent from the source(s) into the dispensing manifold(s) whereby liquid reagent is deposited in the wells;
(c) aspirating pump means operably associated with the outlet(s) of the aspirating manifold(s) for creating a vacuum therein whereby liquid reagent is aspirated from the wells;
(d) means for moving the head assembly along its vertical axis;
(e) a table mounted beneath the head and being adapted to carry the assay tray, the table and/or head being moveable horizontally relative to one another;
(f) means for moving the table and/or head horizontally to position the rows of wells in registry with the dispensing tubes and aspirating tubes; and
(g) means for controlling each of the moving means for the head and table, the dispensing pump means, and the aspirating pump means whereby predetermined volumes of liquid reagents are deposited and aspirated from the wells in a predetermined sequence.
In the drawings:
FIG. 1 is a perspective, partly broken-away view of a prototype embodiment of the assay machine of the invention;
FIG. 2 is a partly sectional, partly broken-away side elevational view of the machine of FIG. 1, showing the head assembly of the machine in its upper position;
FIG. 3 is an enlarged sectional view taken in the direction of arrows 3--3 of FIG. 2;
FIG. 4 is a partly broken-away top view of a portion of the machine of FIG. 1;
FIG. 5 is a sectional, side elevational view of a portion of the machine of FIG. 1 showing the head assembly of the machine in its lower position;
FIG. 6 is an enlarged top view of a portion of the assay tray shown in FIG. 1;
FIG. 7 is a sectional view taken in the direction of arrows 7--7 of FIG. 6;
FIG. 8 is a sectional view taken in the direction of arrows 8--8 of FIG. 7;
FIG. 9 is a sectional, partly schematic view of a portion of the machine of FIG. 1 showing the operation of the liquid withdrawal mechanism of the machine;
FIG. 10 is an enlarged top view of a portion of a second embodiment of the assay tray;
FIG. 11 is a sectional view taken in the direction of arrows 10--10 of FIG. 10;
FIG. 12 is a sectional view taken in the direction of arrows 12--12 of FIG. 11;
FIG. 13 is a sectional, partly schematic view corresponding to FIG. 9 showing the operation of the machine to withdraw liquid from the assay tray of FIG. 10.
FIG. 14 is an enlarged top view of a portion of a third embodiment of the assay tray;
FIG. 14a is a sectional view taken in the direction of arrows 14--14 of FIG. 14;
FIG. 15 is an enlarged top view of a portion of a fourth embodiment of the assay tray;
FIG. 15a is a sectional view taken in the direction of arrows 15--15 of FIG. 15;
FIG. 16 is an enlarged top view of a portion of a fifth embodiment of the assay tray;
FIG. 16a is a sectional view taken in the direction of arrows 16--16 of FIG. 16;
FIG. 17 is an enlarged top view of a portion of a sixth embodiment of the assay tray; and
FIG. 17a is a sectional view taken in the direction of arrows 17--17 of FIG. 17.
The tray and machine are especially useful for conventional solid phase immunoassays. It will be appreciated, however, that the invention may be used for assays or heterogeneous reactions that involve physical and/or chemical interactions or reactions between substances other than immunochemicals. The particular trays and machine shown in the drawings are designed to carry out enzyme immunoassays (EIAs), radioimmunoassays (RIAs), chemiluminescent immunoassays (CIAs) or fluorescent immunoassays (FIAs) in which the solid phase is a bead of an immunosorbent material.
The machine, which is best illustrated in FIGS. 1, 2, 4 and 5, is a modification of the machine that is described in copending, commonly owned U.S. application Ser. No. 06/489,866, filed 5 May 1983. Referring to those Figures, the present machine includes two main movable parts: a horizontally translatable table 15 and a vertically translatable head assembly 16. The table 15 is mounted on hardened guide rods 17 by means of slide bearings 18. The table is translated horizontally by action of a stepper motor 19 through a pinion gear 22 connected to the motor and a rack 23 mounted on the underside of the table. Head assembly 16 is mounted for vertical movement on guide rods 24 by means of slide bearings 25. Translation of the head assembly is provided by a stepper motor 21 via a similar pinion gear-rack arrangement or lead screw mechanism (not shown).
Head assembly 16 supports three main subassemblies: a pipette and plunger subassembly 26; a dispensing manifold subassembly 27; and an aspirating manifold subassembly 28. These three subassemblies are the parts of the machine that introduce liquids into the assay tray wells and withdraw liquids therefrom. The pipette subassembly is an optional feature of this machine and is useful when it is necessary or desirable to be able to transfer precise amounts of liquid reagents to and from the tray. The details of the structure and function of the pipette subassembly are described in said copending application Ser. No. 06/489,866, which disclosure is incorporated herein by reference. In view of this, only a brief description of this subassembly follows.
The pipette subassembly includes a series of pipettes 29 (FIG. 2) that are arranged in a row transverse to the axis of translation of table 15. The pipettes are removably attached to the head assembly by means of a mounting block 32. A plunger mechanism 33 is mounted on the head assembly for vertical movement relative to the pipettes. The plunger mechanism includes a series of plunger rods 34, one being disposed respectively within each pipette. All of the rods are mounted on a common actuator bar 35 for concurrent vertical movement. The bar 35 is translated along guide rods 36 by means of a stepper motor 37 and a lead screw drive mechanism 38. Translation of the plunger rods relative to the pipettes changes the internal air volumes of the pipettes, causing a similar liquid volume to be aspirated into or expelled from the pipette tips 45.
The table 15 includes two work stations 39 and 42 for respectively accommodating two trays 43, 44. Tray 43 is a conventional titer tray that includes a matrix arrangement of wells 41 for housing the liquid(s) to be taken up by the pipette tips. Instead of a matrix of wells, tray 43 may be provided with one or more troughs that provide common liquid reservoirs from which liquid may be taken up. Tray 44 at the rear work station 42 can be a tip tray that contains a similar arrangement of receptacles that accommodate disposable pipette tips 45. The tips 45 are inserted onto and engage the respective ends of the pipettes 29 when the head assembly 16 is lowered by the stepper motor 21 after the table 15 has brought one row of tips 45 into registry with the pipettes.
Once the tips are in place on the pipette ends, the table is translated such that the pipettes are above and in registry with a row of wells in tray 43 that contains the liquid to be taken up into the tips. Instead of wells, tray 43 may have a common trough-like receptacle into which the tips are lowered. The tips are then lowered into the wells by translation of the head assembly and the tips are actuated by the plunger mechanism to withdraw a predetermined volume of liquid from the wells. The tips are then withdrawn from the wells by upward translation of the head assembly. As shown schematically in FIG. 5, the table is then translated to bring assay tray 46 positioned at work station 47 of the table below and in registry with the tips. The tips are lowered by downward translation of the head assembly into the wells 50 of tray and the fluid is ejected by actuation of the plunger mechanism of the pipette subassembly. If desired, the pipette operation may be repeated to extract more or other liquids from the titer tray and deposit it/them in the assay tray wells, with or without replacement of the tips, as desired.
Removal of the tips 45 from the pipettes is accomplished with a tip ejector means. The tip ejector means includes a stripper plate 48 that is best illustrated in FIG. 2. The plate has openings that accommodate the pipettes. The plate 48 is connected to and supported by a pair of vertically translatable rods 49 mounted on the head assembly 16. These rods are translated by means of a pair of solenoids 52 mounted on the head assembly. When the solenoids 52 are deactuated, the ejector plate 48 is maintained in its upper position. Actuation of the solenoids moves the plate vertically downward, to push the tips 45 down and release them from their frictional engagement with the ends of the pipettes 29.
The dispensing manifold subassembly 27 is used to introduce liquids into the wells 50 of the assay plate 46 except in instances where it is necessary or desirable to introduce the liquid via pipette tip because of the availability of the liquid reagent or the need for precise volume control. It is best shown in FIGS. 1, 2, and 3. It comprises a manifold block 53 that is mounted on the front of the head assembly such that it extends outwardly therefrom parallel to and above table 15. The block 53 contains a plurality of horizontally spaced, parallel chambers 54 for receiving the various liquids that are introduced into the wells according to the protocol of the assay being conducted. The spacing between the vertical center lines of the chambers is the same as the spacing between the center lines of the rows of wells in the assay tray. Referring to FIG. 3, each chamber has a single inlet in its upper wall fitted with a tubular sleeve 55 and a plurality of equispaced outlets (six in the depicted manifold) each fitted with a liquid dispensing tube 56 that extends vertically through the bottom wall of the block into a chamber 54. Pressure tight conduits 57 are pressure fit over the outer end of each sleeve 55 and extend therefrom to the exhaust ports of peristaltic (or similar fixed displacement pump) pumps 58. The inlet ports of the pumps are connected via another set of conduits 59 (FIG. 1) to containers 62 that hold the liquids that are used in the assay.
If expensive reagents are involved it will be desirable to use alternate assemblies of liquid containers and pumps that minimize dead volume. Such assemblies would involve a minimum of fluid passageways between the liquid reservoir and manifold. For instance self-contained syringe pump units mounted directly on the head assembly and stepper motors to drive the syringe plungers might be employed. Accordingly, as used in the claims the term "pump means" is intended to denote any device or mechanism that exerts the pressure (or suction in the case of aspiration) required to effect the desired liquid transfer.
Aspirating manifold subassembly 28 is the part of the machine that is used to withdraw liquid from the wells of the assay tray as required by the assay protocol. It includes an aspirating manifold block 63 mounted on the front of actuator bar 35 by means of arms 64 such that it is normally located outwardly and above the leading end of the dispensing manifold block 53. The aspirating manifold block contains a single chamber 65 that has a plurality of equispaced inlets each fitted with a liquid aspirating tube 66. These tubes are longer than the dispensing tubes 56 so that when the dispensing and aspirating manifold blocks are in their uppermost positions the distal ends of tubes 56 and 66 lie in a horizontal plane (FIGS. 9 and 13). The spacing between the tubes 66 is the same as the spacing between the center lines of the columns of wells in tray 46. The spacing between tubes 66 and the first row of dispensing tubes 56 along the axis of translation of table 15 is less than the diameter of the assay tray wells so that they may be positioned simultaneously in the same row of wells 50. Chamber 65 has a single outlet fitted with a sleeve 67. A pressure tight conduit 68 is fit over the outer end of sleeve 67 and extends to the inlet port of a peristaltic (or other fixed displacement pump) pump 69. The outlet port of pump 69 is connected via another conduit 72 to a receptacle 73 for receiving liquids withdrawn from the assay tray wells.
FIGS. 6-17a illustrate the structure of the assay tray wells and their relationship to the liquid dispensing and aspirating tubes. The wells shown in FIGS. 6-8 are cylindrical depressions, each of which has the same structure. Each well has a flat, sloping bottom wall 74 that has a diametrical slot 75 in it. The slope of wall 74 and slot 75 both serve to cause the solid phase used in the assay, in this case spheres or "beads" 76, that are placed in the wells to assume by gravity uniform and relatively fixed positions. In this regard, the width of the slot 75 is less than the diameter of the bead so that latter cannot slip down into the slot. As shown the bead always assumes a resting position seated in the slot at the low end of the sloping bottom wall. The bottom of the slot includes a section 77 that is parallel to the bottom wall of the tray and spaced horizontally from the low end of the bottom wall of the well and an upwardly sloping section 78 that is generally beneath the position normally occupied by the bead. Slot 75 has several functions. First, it provides a seat for the bead. Second, it provides a drain or sump for liquid that may be accessed vertically by the aspirating tubes 66 without disturbing the bead. Third, section 77 provides an optically flat surface through which light may be transmitted in a vertical path that does not intersect the bead in order to carry out optical measurements on the liquid contents of the well. It will be appreciated that the portion of section 77 that provides an optical surface need not be flat but may define a lens or filter for focussing or otherwise altering the light passing through it.
The well 79 depicted in FIGS. 10-13 differs structurally from the well 50 of FIGS. 6-9, but functions in the same manner. This well consists of three sections: (a) an upper main well section 82, (b) a centrally located subwell 83 that provides a receptacle for the bead, and (c) a sump section 84. Each of the three sections opens into the other. Upper section 82 forms the mouth of the well and is generally frustoconical in shape, diverging outwardly directly from the interface with and opening of the subwell. Its shape causes the bead to drop into the subwell 83 when the bead is placed in the well. The frustoconical shape of the main section also facilitates the flow of liquid through the subwell during washing steps and provides a sufficiently large liquid surface to permit overhead fluorescent reading of the liquid contents of the well. In this regard, the minimum diameter for FIA reading is approximately 8 mm. The diameter of the main well section at its mouth will usually range between 15 and 20 mm. The subwell is generally cyclindrical in shape. Its diameter is less than the average diameter of section 82 but greater than the diameter of the bead. Standard bead sizes are 4.4, 6.4 and 8 mm. Accordingly the mean diameter of the subwell normally will range between 6 and 10 mm depending on the size bead it is intended to hold. Preferred sizes are: 10 mm diameter for 8 mm bead; 8 mm for 6.4 mm bead and 6 mm for 4.4 mm bead. It has a concave shaped bottom wall. Sump section 84 is in the form of an elongated slot whose width and thickness are less than the diameter of the bead so that the bead cannot fit into it. It is at least as deep as the subwell 83 so that liquid can freely drain into it from the subwell. The bottom wall of the sump is flat and parallel to the bottom wall of the tray. The configuration of the well 79 serves the same function as that of well 50, with an additional advantage. The frustoconical shape of main section 82 and the bead receptacle section combine to serve as a bead positioning means. Sump section 84 provides a fluid drainage area that may be accessed vertically without danger of disturbing the bead. The bottom wall of the sump section provides an optically flat surface for transmitting light through the fluid contents of the well without intersecting the bead. The configuration and size of the subwell permit the bead to be immersed with a relatively small volume of liquid. In this regard the volume of the subwell will normally be 30% or less of the volume of the entire well, usually 10% to 25% of the entire well. In general, the minimum volume of liquid to immerse the above described standard beads in the subwell will depend upon the size of the bead and diameter of the subwell. In the case of the above standard sized beads and using a subwell diameter about 1-2 mm larger than the bead diameter, the minimum volume will be about 200 to 500 .mu.l. This ability to immerse the solid phase in a minimum volume of liquid is important when the assay protocol calls for use of expensive reagents. At the same time the upper liquid reservoir section allows the use of larger volumes of liquid such as are typically used in washing the solid phase.
FIGS. 14 and 14a show a well 93 that has a pair of axial spaced ribs or pillars 94 having sloping flat top surfaces 95. The ribs define a ramp that causes the bead to drop into a bead receptacle section 96 of the well. The space 97 between the ribs is less than the bead diameter and defines a sump area that may be accessed vertically without disturbing the bead.
FIGS. 15 and 15a and 16 and 16a depict trays in which the solid phase positioning means is a plurality of spaced vertical posts positioned strategically in the wells. The well 98 of FIGS. 15 and 15a has three axially elongated posts 99 with sloping top walls 102. The posts are aligned chordally and the distances between the posts and between the posts and the near portion of the cylindrical wall of the well is substantially less than the diameter of the bead. When a bead is dropped into the well this spacing and the sloping top walls 102 cause the bead to be positioned eccentrically in the well. The space between the posts and the near portion of the cylindrical well wall defines the sump area of the well. FIGS. 16 and 16a show a well 103 that has four vertical posts 104. The posts are positioned in the four quadrants of the well and have top surfaces 105 that slope downwardly toward the center of the well. This arrangement of wells causes the bead to be positioned centrally in the well. Sump areas are located between the posts and the cylindrical wall of the well.
FIGS. 17 and 17a depict a well configuration that is similar conceptually to the well of FIGS. 16 and 16a. The well 106 of FIGS. 17 and 17a has four radial ramps or fins 107 instead of the four posts 104 of well 103. The top walls of fins 107 slope downwardly toward the center of the well, again causing the bead to be positioned centrally. Sump areas are located between the fins. The bottom walls of each of wells 93, 98, 103, and 106 are flat so as to provide a flat surface through which a light beam may be passed vertically in a path that does not intersect the bead.
The operations of pumps 58, 69, stepper motors 19, 21, 37 and solenoids 52 are controlled by a microprocessor 85. The microprocessor controls the sequence of operations of each of these elements and thus the interrelated movements of (a) the table on which the assay tray is held (b) the head assembly and the various parts thereof that move vertically, and (c) actuation of the pumps to transfer liquids to and from the wells. When the machine includes means to spectrophotometrically read the liquid or solid phase in the well the microprocessor may also be used to control the spectrophotometer, record the readings made by it, process those readings or correlate them to other data, and generate reports based thereon.
As indicated previously, the apparatus shown in the drawings is designed to carry out EIAs. In EIAs one is typically looking for the presence or quantity of a multivalent antigen or hapten in a body fluid. Alternatively, EIAs may be used to look for the presence of a particular immunoglobulin in a body fluid which is indicative of a particular medical condition. In the former an immunoglobulin that is specific for (binds selectively to) the antigen/hapten of interest is immobilized on the beads. In the latter the antigen is immobilized on the bead surface. Various types of materials are used to make the beads depending upon the nature of mechanism by which the immunochemical (antibody or antigen/hapten) is affixed to the bead surface. In this regard the term "immobilization" is intended to denote the ability to hold the immunochemical by chemical (e.g., covalent bonding, hydrogen bonding) or physical mechanisms (e.g., adsorption, hydrophobic-hydrophilic affinity). Immunosorbent materials such as crosslinked dextran, agarose, silicated glass beads, polyacrylamides, polystyrene, or latex that bind the immunochemical by adsorption are commonly used. It is expected that for commercial distribution the immunochemical-bearing beads will be prepared beforehand, placed in the wells, the wells sealed such as with a strippable cover and the trays packaged for storage or shipment. The wells of a given tray may contain the same immobilized immunochemical or different immobilized immunochemicals if the tray is to be employed for different assays. Color coding or labels may be used to designate the contents of the wells and/or the assay for which they are intended.
An EIA for an antigen/hapten will typically involve the following sequence of steps.
1. Add samples and control reagents to the tray wells (and beads if they have not been pre-loaded)
2. Dilute sample with buffer (this is optional and the buffer may be added to the well before or after the sample is added)
3. Incubate well contents
4. Withdraw samples/controls from wells
5. Wash beads with buffer (typically repeated 2-5 times which involves addition and withdrawal of buffer)
6. Add enzyme-labeled antibody reagent that binds specifically to the antigen/hapten
7. Incubate
8. Withdraw enzyme-labeled antibody reagent
9. Wash beads with buffer (again, typically repeated)
10 Add substrate
11. Incubate
12. Add enzyme deactivator (optional in many assays)
13. Read wells spectrophotometrically. Enzyme activity is related to antigen concentration and is determined by measuring the optical properties of the liquid contents of the wells at the conclusion of the assay (step 13 above) and comparing those properties to standard curves.
The invention machine would perform these steps in the following manner. The assay tray 46 preloaded with immunosorbent beads bearing the cognate antibody to the antigen/hapten is placed on table 15 at work station 47. Buffer is placed in the container 62 that feeds the first of chambers 54 (i.e., the one nearest the aspirating manifold subassembly 28). Enzyme-labeled antibody reagent, substrate, and enzyme deactivator are, respectively, placed in each of the other containers 62. Predetermined volumes of samples/controls may be loaded into a titer tray 43 at work station 39 and transferred therefrom to the wells 50 of tray 46 using the pipette subassembly as described above. After the samples and controls have been loaded into the assay tray wells the table is translated so that a row of loaded wells is below the row of dispensing tubes 56 that communicate with the dispensing manifold chamber that is interconnected to the container 62 holding the buffer. The dispensing manifold is then lowered such that the ends of those tubes are over a row of wells. The peristaltic pump 58 that serves the buffer line is then actuated to pump buffer into the dispensing manifold and out of the dispensing tubes into the wells. The samples/controls in the row of wells being worked on is thereby diluted to the desired concentration. The table is then moved to position the next row of wells beneath the buffer dispensing tubes, and the sequence is repeated to dilute sample/control in that row. Thus the sample/control in all or some of the rows may be diluted.
After the buffer dilution step the tray is incubated under conditions that permit any antigen/hapten in the sample/control to bind to the bead. If desired the machine can be equipped with a heating plate (not shown) that underlies the table 15 so that station 47 can be positioned over the heating plate to control the incubation temperature. Also, the table 15 may be translated to position the tray 46 beneath a shroud 86 to lessen the likelihood of the contents of the tray being contaminated. The table might also be agitated to facilitate mixing of the wells' contents through use of stepper motor drive 19. At the completion of the incubation, the table is translated to move the assay tray into position for the sample/control to be removed from the wells and the beads to be washed. In this operation the row of wells to be worked on is positioned beneath the aspirating tubes 66 and the first row of dispensing tubes 56 (which dispense buffer). (As indicated previously the spacing between the row of aspirating tubes and the first row of dispensing tubes is such that both rows of tubes may be received in the same row of wells). The aspirating manifold is translated downwardly to move the aspirating tubes down into the sump area of the wells (as shown in FIGS. 9 and 13). The dispensing manifold is concurrently translated downwardly to move the buffer dispensing tubes over the same wells. The peristaltic pump 69 is then activated, causing the spent sample/control to be aspirated into the aspirating manifold, and pumped therefrom into waste liquid receptacle 73. The peristaltic pump in the buffer line is then activated to pump buffer (wash liquid) into the wells, thereby washing the bead. Waste wash buffer is withdrawn by reactivating pump 69. By alternating activation of the aspirating pump 69 and the buffer line pump as many wash cycles as desired may be made. The aspirating tubes are then withdrawn from the row of wells and the table is moved, if desired, to carry out wash operations on succeeding rows of wells.
Upon completing the washing, the table is translated to position a row of wells beneath the dispensing tubes 56 that communicate with the chamber of the dispensing manifold that is interconnected to the container 62 that holds the enzyme-labeled antibody reagent and the dispensing manifold is translated downwardly to lower those tubes into the wells. The pump in the enzyme-labeled antibody line is then activated to cause a predetermined volume of that reagent to be deposited in the wells. The dispensing manifold is then raised. This sequence is repeated to deposit labeled reagent in other rows on the tray. After the labeled reagent has been added to the wells, the tray is again incubated. Following incubation spent labeled reagent is withdrawn and the beads are washed using the same procedure that was used to withdraw spent sample/control and wash the beads previously. Substrate is then added to them by translating the table, lowering the substrate dispensing tubes into the wells and activating the pump in the substrate line. Following another incubation, enzyme deactivator is added to the wells by moving the wells into position beneath the deactivator dispensing tubes, lowering them, and activating the pump in the deactivator line.
After deactivation the optical properties of the liquid contents of the wells are read to measure enzyme activity. This may be done by removing the trays from the machine and reading the wells with standard EIA reading equipment. Alternatively, the reading step may be carried out without removing the trays as follows.
Referring to FIG. 2, the machine is equipped with spectrophotometric means for carrying out the reading step without removing the tray. This means includes a light beam generator 87 that underlies the table 15 that is capable of generating beams of light 88 and a spaced, photoelectric receiver 89 mounted on the underside of shroud 86 opposite generator 87. The table has an aperture 92 in it at station 47 such that the bottom wall of the tray beneath the wells 50 is exposed. In operation, the table is translated to position the optically flat sections of the wells' sumps between the generator and receiver, the generator is activiated to pass the beam upwardly through the liquid contents of the well to the receiver. The light energy received by the receiver may be recorded and analyzed by conventional means. The operation of the generator and receiver and the recording and analyzing of the energy received by the receiver may also be handled by microprocessor 85. Microprocessor 85 may be linked to data storage means (e.g., disk drives) and read out means (CRT displays, printers) to store the data and display it or the reports generated using it.
Other modifications of the above described embodiments of the invention that are obvious to those of skill in the fields of mechanical engineering, immunoassay devices, robotics, and related fields are intended to be within the scope of the following claims. Such modifications may include, without limitation, altering the machine so that the head assembly is horizontally movable and the table is fixed horizontally or both are moveable horizontally. Also the horizontal movement may be rotational rather than linear (e.g., the trays could be carried on a carousel with wells arranged in radial or circumferential rows). The machine might also be altered to carry out FIAs automatically. In such an embodiment the assay reading means would comprise a fluorometer which would read the fluorescence of the liquid contents of the wells. The predetermined location of the bead in the well would enable accurate and reproducible direction of the excitation energy and reading of the resulting emission energy. Further, well configurations that are conceptually similar to the embodiments described in the drawings are readily apparent, such as configurations that comprise a bead receptacle well and a separate horizontally spaced sump that is interconnected via one or more channels to permit fluid to move freely between the bead well or wells that contain screens or projections other than the illustrated posts, ramps and fins.
1. An assay tray for carrying out an assay wherein a particulate solid phase is contacted with one or more liquids said tray comprising a plate having a plurality of vertical wells therein for receiving an equal number of solid phase elements and one or more liquids, said wells having
- sump means in the bottoms thereof which are unobstructedly accessible from the tops of the wells such that liquids may be essentially completely aspirated from the wells by aspiration apparatus lowered into from the tops of the wells;
- a solid phase positioning means in and the sump means comprising an integral part of the structure of each of said wells for positioning a solid phase element in each well such that when said wells are in an upright position a solid phase element placed in any one well in contact with said solid phase positioning means always moves under the influence of gravity to a predetermined location in said any one well away from said sump means.
2. The assay tray of claim 1 wherein the solid phase elements are generally spherically shaped beads said solid phase positioning means comprise a flat sloping bottom wall in each well and the sump means comprise a slot in each sloping bottom wall that runs along the axis of slope, and wherein the sump means comprise walls that are transparent such that a light beam may be passed through the sump means.
3. The assay tray of claim 1 wherein the solid phase elements are generally spherically shaped beads, and said solid phase positioning means comprise a plurality of spaced axial posts within each well which posts are spaced so as to cause a bead to be positioned at a predetermined location at the bottom of each well.
4. The assay tray of claim 1 wherein the solid phase elements are generally spherically shaped beads, and said solid phase positioning means comprises a plurality of spaced ramps within each well that are spaced so as to cause a bead to be positioned at a predetermined location at the bottom of each well.
5. The assay tray of claim 1 wherein said predetermined location is generally in the center of each well
6. The assay tray of claim 1 wherein each sump means comprises at least one wall that has an optical area through which a beam of light may be passed along a path that does not intersect the predetermined location whereat a solid phase element is positionable by means of the solid phase positioning means.
7. The assay tray of claim 6 wherein the optical area is optically flat.
8. The assay tray of claim 1 wherein the solid phase elements are generally spherically shaped beads said solid phase positioning means comprise a sloping well sidewall in each well and a subwell in each well for receiving the beads, and the sump means comprise a vertical slot in each sidewall that opens into the subwell whereby liquid may drain into the slot from the well and subwell when the wells are in an upright position.
9. The assay tray of claim 5, wherein each slot has an optical area through which a beam of light may be passed along a path that does not intersect the predetermined location whereat a bead is positioned by means of the solid phase positioning means.
10. An assay tray for carrying out an assay wherein a particulate solid phase is contacted with one or more liquids, said tray comprising a plate having a plurality of vertical wells therein for receiving an equal number of solid phase elements and one or more liquids said wells each having:
- (a) a subwell means therein for guiding a solid phase element to a predetermined location in each well and holding it in said predetermined location when the wells are in an upright position:
- (b) an upper mainwell section; and
- (c) sump means in the bottoms thereof located away from said predetermined location such that said sump means may always be used to essentially completely aspirate liquids by aspiration apparatus lowered into said sump means from the tops of said wells, said sump means comprising lowest portions of the well bottoms.
11. The assay tray of claim 10 wherein the volume of each subwell section is less than about 30% of the volume of its respective well.
12. The assay tray of claim 10 wherein the volume of each subwell section is about 10% to about 25% of the volume of its respective well.
13. The assay tray of claim 10 wherein said
- sump means are spaced from said subwell sections such that liquids may be withdrawn from the wells without disturbing solid phase elements located in the predetermined locations.
14. The assay tray of claim 10 wherein said subwell sections are generally cylindrical and open to the upper mainwell sections and wherein the upper mainwell sections are generally frustoconical and diverges outwardly and upwardly from the openings of the subwell sections and terminate at open mouths in the upper mainwell sections.
15. The assay tray of claim 14 wherein the diameter of the subwell sections is in the range of about 6 to 10 mm and the diameter of the upper mainwell sections at the open mouths thereof is in the range of 15 to 20 mm.
Referenced Cited
U.S. Patent Documents
|August 1974
|Drozdowski et al.
|November 1975
|Bak et al.
|January 13, 1976
|Beall et al.
|February 17, 1976
|Lanier et al.
|October 12, 1976
|March 15, 1977
|Finter et al.
|March 15, 1977
|October 11, 1977
|May 2, 1978
|April 3, 1979
|Suovaniemi et al.
|November 4, 1980
|December 23, 1980
|May 5, 1981
|Mandle et al.
|October 13, 1981
|March 2, 1982
|Halbert et al.
|March 16, 1982
|Suovaniemi et al.
|May 10, 1983
|Kutsusawa et al.
|September 20, 1983
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Patent History
Type: Grant
Filed: Oct 1, 1984
Date of Patent: Jul 21, 1987
Assignee: Cetus Corporation (Emeryville, CA)
Inventors: Larry J. Johnson (San Jose, CA), Stephen R. Coates (Orinda, CA), Rueyming Loor (Hercules, CA)
Primary Examiner: Barry S. Richman
Assistant Examiner: Michael S. Gzybowski
Attorneys: Thomas E. Ciotti, Ronald A. Fish, Leona L. Lauder
Application Number: 6/656,234
International Classification: G01N 3350; G01N 3500;


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The Oldham County History Center will be hosting an archaeology investigation from 11 a.m. to 2 p.m. on Sunday, May 22. This is a continuing investigation at the Gatewood Plantation, located outside of Bedford in Trimble County.
As wife of the 16th President of the United States of America, Mary Todd Lincoln led a fascinating life. She was privy to the White House and all it contained and was often criticized for spending too much money. Many assumed she came from a slave-holding Kentucky family and therefore had Confederate sympathies. Others felt her partnership with Lincoln was a betrayal of her Southern heritage. But Mary Todd had a life before and after she met Abraham Lincoln at a dance in 1839.
The Oldham Chamber and Economic Development office is currently accepting nominations for the 2015 Oldham Countian of the Year Award. Applications/nominations have been distributed and are available by contacting the Chamber office at 222-1635 or visiting their website at www.oldhamcountychamber.com. Applications must be submitted by Friday, Oct. 2.
If you take care of a home and/or family, then you're a homemaker. Men and women of any age and background can be homemakers. Today, many also work outside the home to help provide financially for their families.
Spend a few hours on a downtown walking tour of La Grange and learn things you never knew about the quaint town beside the railroad tracks. Join Dr. Nancy Stearns Theiss, Ph.D., executive director of the Oldham County History Center, as she leads an informative and fun stroll through La Grange, pointing out various highlights of the town's history.
A 12-person crew from Boy Scout Troop 28, chartered to Kavanaugh Life Enrichment Center in Crestwood, recently returned from a 12-day expedition through the Sangre de Cristo Mountains at Philmont Scout Ranch in Cimarron, New Mexico.
Philmont covers 214 square miles of wilderness. The crew climbed peaks as high as 11,700 feet and hiked over 70 miles.
Presented by the Oldham County History Center, 11:30 a.m.-1:30 p.m. on June 26
The tiny town of Westport has a huge history that dates far back, long before Oldham County became a county in 1824. Native Americans once inhabited the area before the town began as a land grant bestowed upon Elijah Craig for a Treasury Warrant on May 22, 1780. Some years later, on April 25, 1796, Craig sold the land to Joseph Dupuy and Harman Bowman for 450 pounds. It was a solid investment that would produce a once flourishing river town on the banks of the Ohio River.
Jo Ross has spent 48 years in the fashion industry learning the ins and outs of fast-paced runway shows, working internationally to produce shows in places such as Peru, Prague and Mexico, reporting on fashion for local newspapers and TV stations and curating costume exhibits for many local museums. In all that time she has learned a wealth of information she often shares with others through special programs relating to the textile industry.
By Taylor Riley
Hermitage Farms in Goshen will be the scene of the Hermitage Grand Gala, a star-studded Derby Eve party this Friday, May 1.
The party is a joining of Laura Lee Brown and Steve Wilson, founders of 21c Museum Hotels, and Junior Bridgeman, President of Bridgeman Foods.
Copyright © 2019
The Oldham Era is your source for local news, sports, events, and information in Oldham County and LaGrange, Kentucky, and the surrounding area.


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A lot of time newer is not better; other times it is. We all know that we like the new gadgets and gizmos to come out on the market and some people habitually "upgrade". We all know someone who has a pile of last year's good ideas or has a small warehouse full of items on ebay that are no longer in use. Sometimes I look at a new advertised piece of gear and think; I would like to have that. I start thinking; yeah; I could really use that. Then inevitably my logical side kicks in and I ask the important questions like; do I really need this? Will this improve my life any? Can I get by with something I already own? The answer most of the time is "No you don't need this".
I guess I am not a very good capitalist because I find that I do not spend very much on gear. The odd times I do though; especially if I can find something that will replace two or three things that I would normally take on a trip.
Recently I bought a Leatherman Skelitool multi-tool because it reduced my canoe fix-it kit to a couple of ounces instead of a bag filled with small tools. The problem is that the society we live in says that my purchase is not enough. Shortly after releasing the Skelitool, Leatherman released the Skelitool CX with "New Carbon Fibre Handles".
But what is wrong with my old one? Well the answer is; nothing. Problem is that all the cool kids that bought the new one can look at me and point, laugh, make fun of me and feel good about themselves because their multi-tool is newer then mine and they have a higher status in the tribe than me. They can point to how inferior a person I am because I don't own the latest and the greatest.
The peer pressure and societal pressure can be immense. That is how our society works however and we are taught that this is the way it should be from the time we are kids. We get the Dora the Explorer lunch box which works fine and then one day some girl like Hannah Montana instead and all of a sudden Dora is not cool any more. You have to ditch that Dora lunch box or you will be ostracised by everyone. The overwhelming message in conform or be crushed.
What happens though when the new stuff is just pure crap? Unfortunately better marketing wins the day. Anyone remember VCR and Beta? Not to worry though because six months down the road that crap item will be replaced too.
I don't really have the money required to upgrade all the time and sometimes I get thinking that this may limit the sort of outdoor experiences that I might have. Then I remember that people have been living outdoors for a hundred thousand years; it has just been the last sixty that companies have been trying to sell me things to make my life better. Has much of what they are selling actually made life better in the outdoors? Well yes; it is hard to argue that Royalex is not better than cedar canvas canoes for beating around in the rocks. But it could be said that before plastics people just did not beat canoes around in the rocks, they lined, waded, and portaged. How fun is that? Still I do like my plastic canoes.
Much of the gear that I have tried has not survived the test of time. When something is ready for the garbage I tend to disassemble it and save buckles, straps, and the useful parts for some of my make it yourself projects. I am constantly using parts of dead gear to fix up or build new gear. I save old lawn chare bags to make rescue rope bags out of, and buckles and clips all have a thousand uses. I am not contributing to that capitalist economy I know, but waste not, want not my grandmother always said. She grew up in the great depression so she probably knows a thing or two about wanting and not having.
Over the years I have learned to make a lot of gear out of necessity. Not everyone can do this but if you can you are fortunate. First of all you control the quality and design and second you can sell the odd one so you can more or less support your outdoor hobbies instead of them costing you money. Another thing I learned out of necessity (from learning to make things) is how to spot quality and that is about the most important thing you can learn. If you get that down, then you can literally save yourself thousands because you will only need to buy gear once and you will pretty much be set for life. The down side is you do not get that addictive rush that comes with spending money on new things. In my experience though wanting is more powerful than having so you will probably be happier in the long run having good gear and just wanting the new junk.
Old Versus New
When it comes to canoes you are looking at a big investment. You are not likely going to get much for under a thousand dollars or pounds as the case may be. You can go lower of course but you get what you pay for. You have to decide if this is an investment you will enjoy five or ten times per year, or if this is something that you will use fifty days of the year. If you are approaching the fifty days a year in a canoe then you are well advised to consider investing a little more.
When it comes to canoe build quality though newer is almost always better but you come to a plateau very quickly in canoe quality and beyond that quality plateau marketing takes over and reality is left behind.
For portaging the light modern canoes cannot be beat but the cedar canvas is really no heavier than a modern plastic boat. Traditional Cedar canvas canoes are wonderful for the right person. The right person is someone who is handy and knows how to use tools and fix things but more importantly has patience. Not only the patience it takes to fix things but the patience not to beat the canoe through rocks in the first place; the sort of person who stops and gets out in the water up to his knees and never lets the canoe touch shore. The sort of person who will store and care for the canoe like it is on loan from a museum. The traditional cedar canvas is also good for the type of person who is just paddling about on lakes and canals and is not really going to beat the heck out of it. Having said that; the cedar canvas canoe is capable of being beaten around, is infinitely repairable, and even in the worst condition can usually be brought back to look like new. While it is not on par with the modern it is an old school piece of equipment that comes close in a lot of ways to modern canoes in the hands of the right person.
I won't even mention aluminum and plastic paddles because we all know what they are for. When it comes to paddles a lot of people swear by carbon fibre and manmade materials. I am willing to bet they have never used their monstrously expensive paddle on a hundred kilometre plus trip with thirty or more portages through the woods where you are so exhausted that nothing has value at the end except a few minutes rest. I bet that they have never used it to dig a water trench to keep the rain puddle from forming under the tent in a storm in the dark. Chances are they have not tossed it in the back of a pickup truck to be bounced down a forty mile dirt road.
They may be just the thing for sprinting across the lake but they are unrepairable on a trip, will give you blisters, and any slight damage will give you splinters or cuts. They are light as a feather and have their place but they are not the part of the average canoe user's experience. Traditional wooden paddles are in all ways superior for canoe tripping and paddling purposes period. They are repaireable, can be reasonably light, and you can generally beat the crap out of them and they only look better for it. You will not win a race with them against a determined racer with a carbon synthetic, but the racer will not follow you into the wilderness very far. Also they will need to own all sorts of accessories like paddle bags, paddle gloves, and will spend more time babying their paddle than using it. A lot of people like their modern paddles though.
Well there are no old school PFD's so it is hard to make a comparison. The older style ones that came out before kayaking was popular used to come down and cover the kidneys and keep them warm. The kayaking crossover PFD's are scalloped out for the back of the seat and for the spray skirt. They are not as good but it is what most of us are stuck with. If you can find an older style canoe PFD you should hang on to it; especially if you paddle in cold weather.
Well I really like the modern footwear. I got myself some military type dive boots called Amphib Boots and I really like them for the spring whitewater season and for summer on rivers with a lot of wading and lining. The problem is that they are wearing out all too fast because they are made from all synthetics. A lot of people accept this and when something wears out they just get a new one. I begin to ask; is there something that will not wear out? I picked up some LL Bean Maine Hunting boots and they come with a lifetime guarantee. If they wear out I get a new pair. They have been around for a century and are still made so they have obviously they got something right. Most people have been conditioned that they need to buy footwear every year or so, and are shocked to hear that a pair of boots will last a life time. My great granddad had LL Bean boots in the 40's and they are still as good as my new ones.
Modern feet are used to a level of comfort and support that our grandparents were not used to. At one time shoes were made to fit either foot. Things have advanced considerably but it is pretty hard to find anything wrong with moose hide moccasins or Bean Boots for canoe tripping. I would have to say that some old school footwear is superior to its modern counterparts but not all. The modern stuff is in some cases a little more comfortable, lighter, and better for wading and rock hopping but it just wears out too darn fast. For me this edges the old stuff out on top by just that much. But now I have Gortex liners for my Bean boots and get the best of both worlds.
I have worn out a lot of outdoor clothes. I love the modern quick dry fabrics in the summer but sometimes they do not last more than two hard trips. My old flannel and wool stuff has been with me seemingly for a lifetime. I find that a mixture of old and new is the best compromise on a canoe trip. The lighter modern stuff is good for paddling and portaging while the wools are better at night and for doing camp chores. Trips in the autumn though you can just leave the moderns stuff behind and go with the old school stuff and that certainly lightens the load a bit. My experience with some modern fabrics leads me to believe that they are excellent for other parts of the world, just not mine. It seems that Gortex stops working if it is the wrong temperature or does not work at all if it gets dirty. I am not sure what wilderness you can travel in and not get dirty? I have found that the best waterproof fabric for me is proofed canvas. It is bulky and heavy though so the modern nylon gets used instead in spite of being inferior. The lightness and packability brings it out on top. The waterproofed canvas is prone to abrasion wear though more so than untreated so in spite of how good it works, modern is just better.
I am pretty partial to the modern base layer systems like the Under Armour line up. Wool long underwear is great but I like about the fast drying nature of Under armour. When combined with the old school wool outer clothes they make an unbeatable combination.
While I cannot dismiss modern fabrics all together it has been my experience that they work best when combined with more traditional outer layers because of their increased durability and natural properties. Wool is not Gortex but if you are working hard your body heat will eventually drive any moisture out where it will be shed off and it still works no matter how dirty you get it. For these reasons I conclude that traditional old school clothing still have a place in today's modern world. Just forget being a fashion model for the capitalist finger pointing society if you are wearing granddads hand me downs.
Canvas packs are the subject of a lot of debate. Typically there are two types of people. Most think; why do you want to carry around that old garbage? The others like the look and use it because it was recommended by Bill Mason thirty years ago. Chances are if he was still alive he would still be using it though because there are a third group of people that understand how canvas works. The secret to it is that it breaths. Don't be fooled by companies that say that they wax their canvas or treat it. It will wear out faster. Trust me on this; I have used canvas coats for horseback riding for a lot of years. A waxed canvas coat develops holes inside of 3 years from abrasion and my untreated canvas coat was made in the 1940's.
Canvas packs are infinitely repareable as any tack and harness guy will tell you and if they get wet they will breathe dry with a bit of sun or wind. I would not recommend treating them with anything other than a UV protectant such as 303 Aerospace say every twenty years. They should never be put away wet though because they will start to mold. Another reason some do not like them.
While you have to use dry-bags or a waterproof liner inside them they seem to be infinitely stuffable and can be abused and tossed around with little regard to their wellbeing. They can be prohibitively expensive to some though because they are a quality item in a niche market; a market which few people appreciate or understand. You may notice that the design of the average canvas pack has not changed in the last century. That is because it was perfect then and still is now. Some companies try to sew little pockets on them and add other little gimmicks but really that is just marketing however useful the minor changes are. If they increase the line from two models to ten, then they can sell a few more. Business is business.
Most of the mainstream will not agree but I think old school canvas is far superior for canoe tripping than the modern gear. I have some of both and use canvas exclusively in the canoe. Like the canvas canoe you need to be able to make minor repairs (which is dead easy) and must store canvas packs totally dry to keep them serviceable. At the end of the day though when something goes wrong with a canvas pack I fix it, when something goes wrong with a modern pack I save the buckles and fabric that is salvageable and toss the rest. I hate to toss anything that I paid money for.
At the end of the day
I could go on about tents and stoves and all sorts of gear but I thought I would stick to the basics, the stuff almost everybody uses. I like a lot of modern stuff but I see the value in a lot of old school gear and cannot see it ever being retired completely to the museum. A lot of it has great value and a lot of it is superior in all ways to the modern equivalent. By using the old hand me downs you may get pointed at and laughed at by the cool kids and anyone else that is pretty materials oriented, but you know who you are and what you have. If you need to buy new canvas packs, wool shirts, and quality wooden paddles you will sure pay a lot more because you cannot get them at Walmart or Asda, but at least you will have the knowledge that you can likely pass it on for a few generations and your money is going to a craftsman and not a board of suits with a sweat shop.
If you do go on any of the more arduous type of canoe trips, you can feel safe in the knowledge that you can abuse your gear like you stole it and that it will still work after abusing it for twenty years. At the end of the day it comes down to choice and some people will try to go all modern; others my try to emulate the old-timers and go all antique. Others still will choose the middle path and go with what works regardless. The problem is that it is harder and harder to find some of the old school items that work really well.


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Menus 134-235 USD s.i.
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From December 24th to 25th, from December 31th to January 1st, from May 1st to 1st, Bank holidays, Christmas Holidays.


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Meet Sam Gordon.
She's like a cross between Jamaal Charles, Adrian Peterson and Lance Briggs. If any of those NFL players was a 9-year-old girl.
Sam plays in the Salt Lake City area and scored 35 touchdowns against male competition. Our favorite part of her game: Her vision and yards after contact are ridiculous.
We know who we're taking No. 1 in our Ute Conference Youth Football League fantasy draft.
Follow Gregg Rosenthal on Twitter @greggrosenthal.


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Visions of Oz
Hey WIZARD OF OZ fans!! if you're in LA next month, this is going to be
a really wonderful art show to visit. I'm excited to be participating.
=D Not to mention the fact that the location is just super neat! This is a place I'm dying to visit. Check out "Heritage Square Museum" on google and you'll understand why.
Anyhow, when you visit this link you'll see the list of names. I'm
honored to be included on this list. And I sincerely hope to see you


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Salomon's "5 ways to slay" board arsenal
 PORTLAND, OR (January 27, 2014) – For almost 20 years Salomon Snowboards has aimed to bring together a group of people who are not only some of the best riders in the world, but who help Salomon build some of the best product in the world. For FW2014 we are pleased to present five team setups for freestyle destruction, whether you're riding park, pipe, pow jumps, or back alley streets, each setup is built with its own capacity for carnage- This is 5 WAYS TO SLAY.
All terrain ripper and X Games gold medalist Bode Merrill worked closely with us to develop this back, side and frontcountry creation-The Man's Board and Quantum binding. This setup is not for the faint of heart with the whole 'lotta camber and quadratic sidecut of The Man's Board coupled with the natural fit and enhanced response of the new Crossbow Technology on the Quantum.
Wolle Nyvelt and Taka Nakai's do-it-all quiver killer, the Assassin is teamed up with the brand new Defender binding for a fully loaded weapon of slash destruction. A cross between our most progressive powder and freestyle constructions, the Assassin is a force to be reckoned with when paired with the new Defender-featuring Crossbow Technology for the most reactivity, transmission, and support in the ShadowFit family.
Built for the likes of Jamie Nicholls, Hans Mindnich and younger bro Nils Mindnich, the Sabotage is paired with the Hologram binding for disabling gravity so you can go bigger and badder. The camber Sabotage hands over superior edge control and stability, while the Hologram delivers on the support, flex, and comfort demands of true park bosses like these three.
Arguably today's best urban rider and X Games gold medalist, Quebecois Louif Paradis was our cohort in crime in developing the Villain snowboard and District binding setup for one all-terrain jib machine. The Villain's EQ Rad and true twin shape combined with the response, comfort, and flex of the District binding featuring ShadowFit, means freestyle tweaking and perfect pressing like never before.
Crafted under the watchful eye of Jed Anderson and Chris Grenier, the Salomonder snowboard has forged an alliance with the jib-specific member of the ShadowFit Family, the District binding for a street legal assault vehicle .Featuring the original art of Chris and Jed, a flat profile, and easy press core, The Salmonder has got some serious street cred.
Team tested and approved. Choose your weapon wisely.
Salomon Snowboards has been building and designing innovative snowboards, boots and bindings since 1995 from Annecy, France with marketing headquarters in Portland, Oregon. We challenge the status quo, push boundaries, and critically damage preconceived notions of what riding gear can be. Ride It For Real, Stay Loose and once and for all – Slay Boredom.


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G Suite Training
Interactive training and walkthroughs
- G Suite Training is a Chrome extension, offering simple and interactive lessons to get you and your organization up and running fast with G Suite.
Interactive training and walkthroughs
Using Chrome App management, you can provide the G Suite Training Chrome extension to your whole organization in a matter of minutes.
Access training directly from G Suite applications, including Gmail, Calendar, Drive, Docs, Sheets, Slides, Sites, Forms, Google+, Groups and the Admin console.
When new features become available for G Suite, new training becomes available automatically.
Links to a specific training can be shared within your organization and used as a support tool to answer common questions.
Reporting tools allow administrators to track training usage within your organization.
We have approximately 26,000 corporate employees using G Suite on a daily basis. G Suite Training presented an ideal, scalable training solution that will help our staff become more familiar with using these new tools. We are already seeing increases in employee engagement, productivity and flexibility.
Woolworths Limited
G Suite Training is available in English with limited support for Mandarin, Cantonese, French, German, Japanese, Brazilian Portuguese and Spanish.
You can use Chrome App management, found in your Admin console, to deploy the G Suite Training Chrome extension to your domain. Chrome App management is helpful if your domain has a large number of users or if your users are spread across multiple offices.
Interactive training works on Chrome for desktop, and video-based training works on all modern browsers and devices, including mobile.


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This amazing painting is the master reproduction of "Flower Vendor with Basket 1942" by Diego Rivera. This painting is available in eight different sizes.
More paintings by Diego Rivera
Available Sizes:
1. 8x10 inches (20x25 cm)
2. 12x16 inches (30x40 cm)
3. 16x20 inches (40x50 cm)
4. 20x24 inches (50x60 cm)
5. 24x36 inches (60x90 cm)
6. 30x40 inches (75x100 cm)
7. 36x48 inches (90x120 cm)
8. 48x72 inches (120x180 cm)


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ForumsQuestionsGrid format, priority column
Grid format, priority column
I normally use Multi-Line format, but now that there's a quick switch button I'm considering the value of the Grid format.
One thing I was trying to do with it, though, is make a column really narrow so it only shows a little bit of the data.
For example, I've made the Date Due column narrower than its contents, only showing the month and day and not the year.
However, I can't do the same trick with the Priority column. I really only need to see 2 to know it's High priority, but I can't resize it to be narrower than the column title.
Is this important enough to anyone else to request something to get around this? It's really just a little thing, in a view I seldom use.
Anyways, just putting it out there as an oddity. If the Toodledo folks are tweaking anything else in the Grid view, this might be something else to look at.
Toodledo Founder
Thanks for the suggestion.
Is there any way to set the default for multi-line view to the collapsed state rather than expanded?
As more tasks are added, the multi-line view becomes inundated with info. Collapsing each item individually, in order to get an overview (better than grid view), is repetitive and time-consuming. However, the detail that multi-line view provides is often essential.
Would it not be better if the view opened in the collapsed state, with the ability to then expand the particular item of interest?
Thank you.
Please disregard my note of 35 minutes ago re the default for multi-line view. I found the toggleswitch! Terrific!
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Professor Martin Elliott
Martin Elliott was Gresham Professor of Physic from 2014-2018. He is Co-Medical Director at The Great Ormond Street Hospital for Children, London (GOSH). He led the cardiorespiratory and thoracic transplant teams at GOSH for many years, and established, and is Director of the National Service for Severe Tracheal Disease in Children, the largest such service in the world.
Professor Elliott studied at Newcastle University both as an undergraduate and a postgraduate, and also in the Universities of Southampton and London. He was appointed a consultant paediatric cardiothoracic surgeon at GOSH in 1985 and has been Professor of Cardiothoracic Surgery at UCL since 2004.
His research has covered several domains, including the pathophysiology of cardiopulmonary bypass (developing modified ultrafiltration, now used in most repairs of congenital heart disease (CHD)); the development and application of outcome registries in surgery for CHD (he developed the European Congenital Heart Defects Database, the forerunner of the EACTS and STS registries); the specification of the nomenclature for congenital heart defects (he was founding president of the International Nomenclature Society for CHD); the improvement of teamwork within cardiac care (consulting and working with Formula 1 teams, the airline and hotel industries); and most recently the management of severe tracheal disease in children, including the first homograft and stem cell-based transplantations of that organ in children.
Professor Elliott is passionate about improving the experience for patients and providing them with insight they and their families need to make informed choices. At GOSH he leads on quality and safety, and GOSH has won several prizes for its work in this field. He has given evidence to the Bristol and Francis Inquiries related to the importance of information provision for quality improvement.
He has published over 260 peer-reviewed articles and is in demand as a speaker, having delivered over 300 invited lectures worldwide, including 20 named lectures, to professional and lay audiences. He has taken part in several television documentaries. He reviews for many scientific journals and teaches and operates throughout the world. He has trained many of the world's leading paediatric cardiac surgeons and has been involved in service reform in several countries.
All of Professor Elliott's Gresham lectures can be accessed here.
[The image of Professor Elliott is used courtesy of Becan]


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A sales CRM built for minimum input and maximum output
Immediate access. No credit card required.
So we've designed and refined every element to zero-in and double down on actions that drive deals to close
Pipedrive is rated the easiest sales tool to implement by the leading software review site, G2Crowd. With us there's no 200 page list of functions, and no two day training required. Just log in, fill up your pipeline and start selling.
We believe in selling. It's in our DNA, and we've lived and breathed it for decades. We made a product that makes selling beautiful. It's visual, intuitive, customizable, and our users actually choose to stay logged in constantly. Now that's really beautiful.
From proposals to e-signatures to automatic lead scoring. With 100+ of your favourite apps and integrations available in the Pipedrive marketplace, you can arm yourself with everything you need to sell with certainty.
Immediate access. No credit card required.
The ability to completely customize the data fields and workflow gives me a tailor made CRM. Much different than my past experiences with these types of programs! The mobile app is just as functional as the desktop version, and even has a few little extras.
As a long time user of SFDC and other CRMs, I love the simplicity and intuitive nature of Pipedrive. My favorite feature is the automated pop-up forcing the next step.
It has made our sales department more effective and efficient. It's easy to use and it's constantly updated.
We've recently started using Pipedrive and the changes it has made to the overall efficiency of the sales component of our business is outstanding. If you're looking for a fantastic CRM, don't go past Pipedrive!


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Fresh Friday Finds
1. Watch Ellen today. Yesterday she confronted John MCcain(who couldn't even make eye-contact with her)and rumor has it she may reveal something quite excting (watch out Oprah...)! Special guest and friend of DB, Rory Freedman (Skinny Bitch, Skinny Bitch in the Kitch) will be her guest.
2. For those that missed it, Oprah is vegan (for at least 3 weeks). All I have to say is wow...
3. Sign this petition to help call for a South Atlantic Whale Sanctuary, and help end whaling!
3. Don't Teese Yourself! Ok, fine - Teese yourself. It melts!
4. DB reader, MOR is a former employee of Whole Foods. He calls attention to Wholefoods' not-so-socially-progressive policies. Read Wholefoods Breaks Their Promise.
5. Fujimori is being tried for Human Rights Violations in Peru. He is the first head of state to be extradited to his home country to stand trial for human rights violations. Praxis Institute for Social Justice invites you to visit their bilingual liveblog and follow current developments in the ongoing human rights trial of former Peruvian President Alberto Fujimori.
"Former Peruvian President Alberto Fujimori served from 1990-2000 and is suspected of committing innumerable human rights violations during his decade in office. These allegations have been documented by the Peruvian Truth and Reconciliation Commission (TRC) in its Final Report, published in 2003, and the trial against the former president marks the fulfillment of one of the TRC's most important recommendations for preventing future human rights violations by the state."
For more info:
- BBC News Profile: Alberto Fujimori
- Articles from the New York Times
- Biographical information from Association Pro Human Rights (APRODEH)
6. Non-leather belts can be more than a hemp braid - thank god.
7. On Tuesday, the USDA made it official, implementing a no-downer policy for cattle in the United States. Read the article HERE. 'Downers' are animals too sick or injured to walk or stand, and are typically left to suffer - as veterinary care and euthanasia cost more than most farmers will make on the body of the animal - making it economically undesirable, and ethically disgusting.
7. Cruelty-free, vegan colognes (thanks Peta2):
Abercrombie & Fitch and Fierce by Abercrombie & Fitch Mesmerize Cologne Spray by Avon California North for Men and O2XYGEN for Men by California North Number Six by Caswell-Massey Eau Dynamisante and Eau Tranquility by Clarins Chemistry by Clinique 360° for Men and Perry Ellis m by Perry Ellis Tommy for Men by Tommy Hilfiger Aramis and Lauder Intuition for Men Cologne Spray by Estée Lauder OP Juice by OP REGENCY™Cologne by Merle Norman
Joshua's Vegan Baked Mac & Jeeeeze Casserole
What You'll Need:
- 1 Block Silken Tofu
- 1 Block "Follow your Heart" Cheddar style vegan cheese (or Teese!)
- 2 Tbs "Earth Balance" or other vegan margerine
- 3 Tbs Nutritional yeast (more to taste if desired)
- 1 Tbs sugar or other sweetner
- 1 Tbs salt
- 1/4 cup vegan milk (soy, almond, rice, etc make sure it's plain and not "vanilla" falvored)
- 1/2 cup Bread crumbs (make your own by putting crisp toast in a blender)
- 1 - 2 cups of elbow pasta
Directions: Boil pasta. Combine all ingredients aside from bread crumbs and pasta in a blender or food processor. Mix the "cheese sauce" with the boiled pasta. place in casserole dish, and top with breadcrumbs. Bake on 350 for 15 - 20 minutes.
Tip - for a cheesier dish, shred additional vegan cheese over the bread crumbs before baking.


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Each 1/16" diameter cable has machined and threaded chrome-plated brass tips (male and female) for quick connection and release. Although the manufacturing process is designed to accommodate keys and card holders, most of the cables can withstand much more stress but the manufacturing cost of testing each cable would more than quadruple the cost and selling price.
Accordingly we test received shipments to arrive at average strength ranges only. Our tests indicate the following: if you want resistant strength (RS) above 5 lb the odds are very good; 97% of the cables will be above 8.5 lb RS. 88% of the cables will average 26.4 lb to 90.5 lb, and 9% of the cables will average 189.6 lb.
Offered in packages of five.