EP0248882A1 - A disposable microtiter stick - Google Patents

A disposable microtiter stick

Info

Publication number
EP0248882A1
EP0248882A1 EP87900414A EP87900414A EP0248882A1 EP 0248882 A1 EP0248882 A1 EP 0248882A1 EP 87900414 A EP87900414 A EP 87900414A EP 87900414 A EP87900414 A EP 87900414A EP 0248882 A1 EP0248882 A1 EP 0248882A1
Authority
EP
European Patent Office
Prior art keywords
wells
stick
well
microtiter
per
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP87900414A
Other languages
German (de)
French (fr)
Inventor
Wayne M. Henry
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
ALERCHEK ASSOCIATES dba ALERCHEK
Original Assignee
ALERCHEK ASSOCIATES dba ALERCHEK
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by ALERCHEK ASSOCIATES dba ALERCHEK filed Critical ALERCHEK ASSOCIATES dba ALERCHEK
Publication of EP0248882A1 publication Critical patent/EP0248882A1/en
Ceased legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5085—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above for multiple samples, e.g. microtitration plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • G01N33/5304—Reaction vessels, e.g. agglutination plates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing

Definitions

  • This invention relates to medical apparatus in general and more particularly to a disposable microtiter stick for performing immunochemical tests.
  • the micro- titer stick is an elongated molded polystyrene strip, with a series of evenly linearly-spaced, shallow lensed bottom wells for receiving the fluids to be tested and has a handle extending from one end by which the stick can be grasped.
  • the stick is taken by . its handle and by a snap of the wrist the wells are easily and quickly emptied.
  • a raised border upon the top surface surrounds the wells and permits all the wells to be easily filled together by simply flooding the top surface area about the wells with the fluids. Both the filling and emptying of the wells is done without the need for expensive filling and decantation equipment.
  • the well lense bottom portions are transparent while the well walls are frosted to eliminate optical color interference between adjacent wells and to increase in effect the available work surface area versus smooth optical surface throughout the well interior.
  • the flat bottom surface of the stick is recessed to keep the bottom lense portions form being scratched as when the sticks are rested or slid upon a supporting surface. Further, these sticks are disposable with the completion of any one test.
  • present immunoassay test stick has a series of optically-shaped and frosted test wells except for their -lense portions and a handle by which the stick can be picked up to empty the wells of their test fluids.
  • the top surface of the stick has a boundary border around the wells so that all of the wells can be filled at once by a controlled flooding of the top surface area.
  • One big advantage of the present stick is that it is convenient to use and requires little or no instrumentation to fill and empty in the performance of the immunoassay tests.
  • a further advantage is that maximum capacity of each well is but 100 milliliters and automatic measured dispensing is easily achieved in one step by simply flooding the wells within boundary area with the test fluids.
  • the invention resides in a microtiter stick for use in performing immunochemical tests to determine the extent of the allergy content of a blood speci ⁇ men by chemical treatment of the specimen. It comprises a transparent plastic like body having an affinity for protein.
  • the body is elongated and has a flat top surface with a plurality of small shallow wells in the surface.
  • the wells are aligned in a series along the body in a close proximity to one another.
  • Each of the wells has an optically concave surface and the remainder is frosted to lessen color interference between wells and to improve its surface for the bonding of protein and chemical residues to the surface.
  • a handle projects from one end of the body portion of the stick by which the stick may be grasped and flipped to empty the wells of their contents when the tests have been per ⁇ formed.
  • the body has a flat bottom supporting surface and is recessed provide portions of the lenses of the well. These lense portions are spaced from the- flat bottom to protect the len ⁇ e portions from being scratched when the stick is placed on a supporting surface.
  • the upper surface of the body of the stick has a boundary border in the form of a wall surrounding the flat top surface, the wall being located in close proximity to the well openings so that the wells may be simulataneously filled by controlled flooding of the top surface with specimen or rea- gents used in testing.
  • the stick is of a length sufficient to span the sides of a standard tray and has an extension at one end of accomodate an up ⁇ standing small projection at one side of the tray with the handle extending from the opposite side of the body of the stick over the opposite side of the tray.
  • the body of the stick is notched at the end opposite the handle to accomodate spaced inwardly extending projections on the opposite side of the tray while the handle portion is rested thereon.
  • Figure 1 is a perspective view of a prior microtiter strip now used in performing immunoassay tests, and the like.
  • Figure 2 is a similar perspective view of the present invention in the form of a stick that has shallow wells and a handle by the use of which the stick can be flipped to empty the shallow wells of their fluid contents at times while carrying out the immunoassay tests.
  • Figure 3 is a fragmentary perspective view of both the prior and present pieces assembled together in a tray used to support the pieces while carrying out the immunoassay tests.
  • Figure 4 is a perspective view of the prior microtiter strip and the present stick with illu ⁇ stration being made of the manner in which their wells are filled from a squeeze bottle, the prior strip wells -having been individually filled while shallow wells of the present stick may be filled by flooding the top surface of the stick.
  • Figure 5 is a top plan view of the present welled test stick shown in actual size.
  • Figure 6 is a longitudinal sectional view of the present stick as viewed on line 6-6 of Figure 5.
  • Figure 7 is a bottom plan view of the present stick.
  • Figure 8 is an enlarged end view of the stick as viewed upon its handle end.
  • Figure 9 is an enlarged cross-sectional view taken on line 9-9 of Figure 5.
  • Figure 10 is an enlarged end view of the other end of the stick.
  • Figure 11 is an enlarged fragmentary longi ⁇ tudinal sectional view showing the interior of the wells, the raised boundary border, and the lensed well bottom portions as viewed also on line 6-6 of Figure 5.
  • Figure 12 is an enlarged fragmentary bottom plan view looking upon the bottom of the stick and into the lense bottom recesses.
  • Figure 13 is a perspective view illustrating the manner in which the well contents of the stick are dumped as by grasping the stick by its handle and with but a snap of the wrist dumping its con ⁇ tents into a sink drain rather than by using costly apparatus for decantation of the individual wells.
  • Figure 14 is a perspective view of a photo ⁇ sensor arrangement for reading the residue showing upon the well lense portions at the completion of the tests.
  • the wells When the blood sample of the patient is added and the excess blood subsequently decanted, the wells washed and with a tracer reagent, the wells will be colored in a degree to determine the extent that the patient is allergic to these contagions. With no coloring arising in any well, the patient will be regarded as immune to the particular allergy antigen of the one well.
  • Prior well strips 15 shown in Figures 1, 3 and 4 have been formed of transparent plastic and have a top flat strip 15' with a series of raised openings 16' from which depend cylindrical or test tube-like wells 16 that extend some one half inch in depth below the top flat strip 15'.
  • the deep wells are interconnected with one another at their sides and their bottoms are closed by flat circular portions 16" upon which the residue results from the test will be observed.
  • One end of the top flat strip 15* is extended at 17 and has a small hole 17' therein to fit over a small upstanding projection 19' of a tray 19 normally required for the support of the prior strips 15 to prevent spilling of their well contents.
  • top flat strip 15* is retained on the tray 19 at the opposite side between small inwardly-extending projections 19" that will lie astride end well portion 16 so that the well strip 15 is held against sliding displacement along the tray 19.
  • a small window area 21 is provided in the opposite end extension 22 through which a raised indicia 23 on the tray frame may be read while the welled strip 15 over-lies the tray 19.
  • a top hold down frame e 20 is placed over the collected strips 15 and such present sticks 25 which can be similarly assembled therewith upon this standard tray 19 to hold down all pieces against vertical displacement therefrom.
  • the prior deep welled strips 15 have to have their wells 16 filled individually with measured quantities of reagent or specimen as by the use of a pipette and that is a time consuming task. Decan- tation of the well 16 becomes a similar task and costly laboratory evacuation apparatus is required. Laboratory surroundings have had to be more or less available by the physician for the carrying out of immunoassay tests with these prior welled strips 15.
  • These prior deep welled strips 15 can be placed on a flat supporting surface, but because of their height they cannot be placed with safety while having liquid contents in them. They are easily upset and thus are dependent upon the tray 20 for safe support. Thus, for filling and decantation -. ' operations they need to be assembled between the tray parts 19 and 20.
  • the flat bottom plates 16" of the cylindrical wells 16 of the prior strip 15 make for reading of the test results as to color with the projection of light from through the well bottoms.
  • Such flat bottom plates 16" make for the existence of often unclean circular corners against their cylindrical side walls and significant capillar effects arise thereby to reduce the accuracy of the readings.
  • both the prior and present pieces are made of polystyrene, a plastic well known to have an affinity for capturing antigen and antibody proteins and to enhance the separation of solid matter during the solid phase cycles on pre-coating the stick wells and the performing of the immunoassay tests.
  • the prior flat bottom well plate 16" of the prior strip 15 while having the same affinity for proteins takes on a heterogeneous coating preparation due to the diffi- culty in cleaning of the corners of the flat bottom plate 16" with its cylindrical wall and a hetero ⁇ geneous specimen assay will result.
  • the present stick has been designed to overcome such deficiencies to reduce labor in the use of the stick, facilitate the procedure of carrying out of immunoassay tests, lessen the costly laboratory equipment needed in the carrying out of the assay tests as with the prior microtiter strips 15, and finally to make the procedure more adaptable to physicians and laboratory personnel.
  • the present microtiter stick 25 principally differs in construction from the prior strip 15 by the provision of but shallow well depressions 26 fashioned in a solid plastic body that are bottomed with optically transparent concaved lense portion 26' that conform to the rounded well bottom. On projecting light upwardly through these lense portions 26" onto a photo-sensor 35 see Figure 14, the solid material on the lense portion will be focused on a plane and be more accurately read with the photo-sensor instead of merely sensing through an unfocused plate bottom 16" of prior strip 15. By the focused lighting arrangement a better color interpretation will be obtained.
  • the present microtiter stick 25 is about the same size as the prior art strip 15 and can be placed therewith in the standard two-part try 19-20, see Figure 3.
  • the present stick has an end lip 27 with a small hole 27' therein for assembly over the small upstanding projection 19' on the tray bottom part 19.
  • the other end of the present welled stick 25 instead of a mere lip extension has an elongated handle extension 28 by which the stick 25 is handled at times of being placed upon the tray 19 or removed therefrom to empty its contents.
  • This handle is also conveniently used to label the stick for patient identification purposes deemed critical for retaining the integrity of results.
  • the top tray part 20 is cutaway at 20' on one side to accommodate the handle extension 28. To retain the handle end of the stick against lateral displacement the bottom of the stick is placed between two inwardly-extending projections 19" of the tray 19.
  • the depending body of the stick has spaced vertically-extending re ⁇ Memos 29 and 30 under the handle extension 28 to respectively receive the small inwardly-extending frame projections 19" as the stick 25 is laid down upon the tray 19.
  • the handle 28 can be easily grasped to lift the stick 25 from the tray 19 and this is often done many times during the assay to rid the shallow wells 26 of their contents.
  • the wells 26 being shallow decantation devices are unnecessary to empty the wells.
  • the stick 25 can be lifted and with a snap of the wrist over a sink drain 31 in the manner illu ⁇ strated in Figure 13, the wells are quickly and completely ridded of their contents. All wells are at once emptied with the flip of the stick. Imple ⁇ ment decantation of the individual wells is there- fore not needed.
  • An upstanding boundary border 31 surrounds all of the wells 26 running along the sides and across the ends of the stick to permit flooding of the top surface of the stick for effecting simultaneous filling of the wells 26 with unmeasured quantities as from a squeeze bottle as shown in Figure 4.
  • Each well 26 holds 100 mi ⁇ roliters, suitable in volume amount in drops for assay testing, and to obtain full colorization of the well surface.
  • the resul ⁇ tant residue material will develop color and be read through bottom concaved lense area 26'.
  • the boundary border 31 runs tangentially to the side edges of the wells 26 with no surface area lying therebetween.
  • the border 31 runs to the full extent of the eight wells and is closed off at 31' on the end projection 27 and at 31" on the opposite handle end 28.
  • the wells are fully confined by the boundary border 31 so that as illustrated in Figure 4 the entire series of wells 26 will by simple flooding with a squeeze bottle 32 or a free flowing device for spilling of the reagent or specimen on the top surface area, so that wells 26 will be instantly filled to measured capacity with little excess overflow, and each well holding 100 micro- liters.
  • a stream of large specimen drops 34 will abound from bottle nozzle 33 to flood the top surface of the stick 25 and instantly fill the wells 26. It should be apparent that this flooding procedure cannot be effected with the deep welled prior stick 15 and as illustrated in Figure 4 the deep cylindrical wells will have had to have been filled separately and individually with the drops 34 from the squeeze bottle 32. Any measured quantity of blood specimen could have been ascertained only by counting the drops 34, short of the actual filling of the wells 16, unless and more often a pipette would have been used to discharge measured quantities into cylindrical wells.
  • prior strips 15 comprising the deep transparent cylin ⁇ drical test tube-like wells lying attached and closed upon one another that the optical readings of one well can be hampered by optical interference of the showing of the other wells so that true readings of the respective wells are not so likely to pre ⁇ vail.
  • the wells 26 are not tubes as with the prior stick 15, but are mere shallow and frosted openings formed with inclinded walls in the stick's solid body and without cylindrical well walls and such that there can be little, if any, optical interference between adjacent wells.
  • the present stick 25 is frosted throughout all of its surface areas except for the relatively small surface area of the well lensed portions 26'.
  • the frosting extends over all sur ⁇ faces, top, sides, bottom and down into the shallow wells 26, except for their lensed bottoms 26'.
  • the wells by themselves will also be given by the frosting therein effective added contact surface area to ensure that adequate coating densities will be obtained for better observation of test results.
  • the affinity action of proteins upon the polystyrene thus has been enhanced by the frosting.
  • the frosting will have first been made upon the molding surface of the stainless steel die parts and such frosting transferred during the molding oper ⁇ ation onto the molded work body.
  • the die parts are frosted by an electroylytic etching procedure or by any other accepted frosting procedures. Corres- ponding frosting upon the work stick is thus ob ⁇ tained from the dies.
  • the lense areas of the dies are smoothly ground toward infinite fineness and its clarity is transferred to the plastic work stick body. From so formed die parts, sticks frosted except from the lense areas are produced ready for use, inexpensively and are disposable • after final use in the carrying out of the assay tests.
  • the sticks will have been pre-coated with specified antigen or antibody proteins prior to their delivery to physicians and laboratories, and ready for use.
  • the present stick 25 has a flat bottom surface in which the area beneath the lense portions 26 are recessed. Such recesses will keep the lense por ⁇ tions from being scratched.
  • the flat bottom surface allows the present stick to be rested upon a flat supporting surface and not easily turned over, thereby permitting its use without the need for tray 19. With the frosting extending over the handle data marks can be made thereon.
  • the present stick will be pre-coated with antigens and antibodies in accordance with the physician's specifications and requirements and varying for different geographical locations.
  • There are different allergy contagions for the different locations which will be predetermined and known to the physician within his given geographical area.
  • Some of the common allergy contagions are pollen from trees, grass and weeds, mold spores suspended in air, on surface and in bread, animal dander, bacteria, dust mites, and so on.
  • the wells of the present stick are pre-coated with any of these antigens, one antigen for each well and such other antigens not named that are available in serum form for the purpose.
  • all wells can be filled simultaneously in measured quantities of the different antigens by a multiple dispensing apparatus upon advancing the sticks thereunder in a step by step procedure.
  • the sticks After the stick wells have been dehydrated and the antigens reacted with the polystyrene surface of the wells, the sticks are made ready for distribution to physicians and laboratory users.
  • the physicians and users will merely supply the patient's specimen and reagents and wash liquids to carry on known steps of procedure until a color reading will be seen of more or less color density of the residue material overlying the frosted well surfaces and on the unfrosted lense areas 26'.
  • the results of the assay test can be readily determined by merely viewing the well for the color intensities but for a roe accurate determination by subjecting the stick to a light source 37, Figure 14, and at the proper focal point the residue upon the lense may be observed with a photo-sensor 35. All the readings will be free of optical interferences between wells due to the wells being shallow and frosted.
  • micro, solid phase enzyme-linked adsorbent assays can be carried out with the present sticks. The veterinarian will also find these sticks handy in carrying out tests with animal fluids.
  • both antigens and anti ⁇ bodies are proteins and that they lock with one another upon being brought together in the assay tests.
  • Each of the wells of the microtiter stick is coated with but a specific antigen or antibody depending upon the nature of the assay to be made. Because of the natural affinity of these proteins to a hydropholic surface such as that of polystyrene, these two proteins irreversibly bind, absorb or combined with the pre-coated surface upon the well walls. By frosting the well wall surface enlarges the availability surface of the well and thereby the capacity and the efficiency of the well surface is increased.
  • the quantity of the antibody so bound depends upon how much there is of the antibody in the body fluid, the amount of the antibody that has been captured on the well wall surface, the affinity of the capture, the proximity of contact between the entire body of the specimen and the coated surface, the homogenity of the exposure of specimen body to the coated well wall and the spacial molecular arrangement of the anti ⁇ gens.
  • suitable animal antibodies having enzymes and have color forming molecules attached to them are added to the well already pretreated with the human blood specimen, and with further incu ⁇ bation, washing and removal of material and then by the use of subsequent corresponding enzyme sub ⁇ strate, the well surfaces will develop color of intensities proportional to the level of antibodies contained in the patient's blood specimen and with the degree of coloring being noted, the physician is immediately advised of the type and degree of the allergy of his patient. The assay will have been finally completed.
  • the degree or intensity of the color is propor ⁇ tional to the amount of antibody that was bound upon the well wall from the body specimen.
  • the prior microtiter strip 15 and its bottom plate 16" has had many optical shortcomings since it has only provided low plate surface area for the specimen and cylindrical wall areas of the well optically finished and not involved in optical observation. Color development has been less easily delineated due to inter-well interference due to such optically clear surfaces but unavailable for the viewing or sensing color development. With the present shallow walls the full surface areas are available for sensing of color development. Further, it should be seen that the present stick has been designed for easy molding. That the present stick 25 is less than one-half the height of the prior strip 15 and can be safely rested upright on a flat supporting surface without the need of a tray and at the same time can be used in the trays along with the prior strips. While various changes may be made in the details construction, it shall be understood that such changes shall be within the spirit and scope of the present invention as defined by the appended claims.

Abstract

Une tige de microtitrage jetable utilisée pour effectuer des tests d'immunoanalyse chimique dans le but de déterminer des allergies et des maladies chez les êtres humains et les animaux consiste en une tige de polystyrène allongée en une pièce à fond plat (25) ayant une série d'alvéoles ou puits peu profonds (26) avec fonds en forme de lentilles et une extension formant poignée (28) s'étendant à partir d'une extrémité de la tige de manière à pouvoir saisir et soulever cette dernière. Les puits (26) sont peu profonds de manière à pouvoir les vider simultanément en prenant la tige par la poignée (28) et à vider tout le contenu des puits de la tige (25) par un petit coup sec du poignet. Un bord frontière surélevé (31) s'étend autour de la surface supérieure de la tige (25) entourant tous les puits (26) de sorte qu'ils se remplissent tous simultanément avec le réactif ou spécimen simplement en inondant la zone de la surface supérieure bordée de la tige (25). Toutes les surfaces de la tige (25) comprenant les surfaces des parois des puits, à l'exception des zones de vision de lentilles (26') à fonds optiquement transparents, sont dépolies. La surface du fond de la tige (25) est plate de sorte que la tige (25) peut reposer sur une surface de support plate mais elle est évidée sous les zones de vision de lentilles (26') de manière à la protéger contre les égratignures.A disposable microtiter rod used to perform chemical immunoassay tests for the purpose of determining allergies and diseases in humans and animals consists of a one-piece elongated polystyrene rod with a flat bottom (25) having a series dimples or shallow wells (26) with lens-shaped bottoms and a handle extension (28) extending from one end of the rod for gripping and lifting the rod. The wells (26) are shallow so that they can be emptied simultaneously by taking the rod by the handle (28) and emptying all the contents of the wells of the rod (25) with a quick flick of the wrist. A raised border rim (31) extends around the top surface of the shaft (25) surrounding all wells (26) so that they all fill simultaneously with reagent or specimen simply by flooding the surface area upper lined upper (25). All surfaces of the shaft (25) including the surfaces of the walls of the wells, with the exception of the viewing areas of lenses (26') with optically transparent bottoms, are ground. The bottom surface of the stem (25) is flat so that the stem (25) can rest on a flat support surface but is recessed below the lens viewing areas (26') to protect it from scratches.

Description

A DISPOSABLE MICROTITER STICK
Field of the Invention
This invention relates to medical apparatus in general and more particularly to a disposable microtiter stick for performing immunochemical tests.
Background of the Invention
In the performance of the chemical immunoassay tests with microtiter pieces, many filling, emptying and washing procedures are done that are time consuming. Prior microtiter pieces have had deep wells with cylindrical vertical side walls that have taken time to fill and are difficult to empty and clean. Expensive devices have been required to separately fill and empty the individual deep wells with reagents and specimens and the tasks have been cumbersome and tedious. Accurate tests have not resulted from using these deep well pieces because full intimate contact of the fluids upon wall surfaces of deep wells of clear plastic do not result and since there is optical interference between adjacent wells of the piece, good test and reliable readings have not always been determined. According to the present invention, the micro- titer stick is an elongated molded polystyrene strip, with a series of evenly linearly-spaced, shallow lensed bottom wells for receiving the fluids to be tested and has a handle extending from one end by which the stick can be grasped. When the emptying of the wells is to be done, the stick is taken by . its handle and by a snap of the wrist the wells are easily and quickly emptied. A raised border upon the top surface surrounds the wells and permits all the wells to be easily filled together by simply flooding the top surface area about the wells with the fluids. Both the filling and emptying of the wells is done without the need for expensive filling and decantation equipment. The well lense bottom portions are transparent while the well walls are frosted to eliminate optical color interference between adjacent wells and to increase in effect the available work surface area versus smooth optical surface throughout the well interior. The flat bottom surface of the stick is recessed to keep the bottom lense portions form being scratched as when the sticks are rested or slid upon a supporting surface. Further, these sticks are disposable with the completion of any one test.
It is the principal object of the present invention to provide a chemical immunoassay test stick with a most favorably shaped and finished surface test well for the residue and color analysis of the fluids being tested.
It is another object of the invention to provide an immunoassay test stick having a series of shallow-bottomed lense wells that are best shaped for effecting rapid, convenient, and accurate performance of immunoassay tests. It is another object of the invention to provide an immunoassay test well stick with a handle extension by which the stick can be grasped and flipped with but a snap of the wrist thereby emptying all wells of their test fluids at one time and done-without the need of expensive liquid decantation apparatus.
It is still another object of the invention to provide an immunoassay test stick in which optical interference between test wells has been eliminated so that more accurate readings will be obtained.
It is further object of the invention to provide an immunoassay test stick in which all of the test wells can be filled with specimens or reagents at one time by simply flooding the top ■ surface of the stick about the well openings.
It is a further object of the invention to provide an imm noassay test stick in which the full interior wall surface of the well is available for immobilizing the antigens or antibodies.
It is a still further object of the invention to provide a immunoassay test stick that does not * require any tray or rack for its support and that may be rested with safety upon any flat surface. Briefly, present immunoassay test stick has a series of optically-shaped and frosted test wells except for their -lense portions and a handle by which the stick can be picked up to empty the wells of their test fluids. The top surface of the stick has a boundary border around the wells so that all of the wells can be filled at once by a controlled flooding of the top surface area.
One big advantage of the present stick is that it is convenient to use and requires little or no instrumentation to fill and empty in the performance of the immunoassay tests. A further advantage is that maximum capacity of each well is but 100 milliliters and automatic measured dispensing is easily achieved in one step by simply flooding the wells within boundary area with the test fluids. For a better understanding of the invention, reference may be had to the following detailed description taken in connection with the accom¬ panying drawing.
Summary of the Invention
The invention resides in a microtiter stick for use in performing immunochemical tests to determine the extent of the allergy content of a blood speci¬ men by chemical treatment of the specimen. It comprises a transparent plastic like body having an affinity for protein. The body is elongated and has a flat top surface with a plurality of small shallow wells in the surface. The wells are aligned in a series along the body in a close proximity to one another. Each of the wells has an optically concave surface and the remainder is frosted to lessen color interference between wells and to improve its surface for the bonding of protein and chemical residues to the surface. A handle projects from one end of the body portion of the stick by which the stick may be grasped and flipped to empty the wells of their contents when the tests have been per¬ formed. The body has a flat bottom supporting surface and is recessed provide portions of the lenses of the well. These lense portions are spaced from the- flat bottom to protect the len≤e portions from being scratched when the stick is placed on a supporting surface.
The upper surface of the body of the stick has a boundary border in the form of a wall surrounding the flat top surface, the wall being located in close proximity to the well openings so that the wells may be simulataneously filled by controlled flooding of the top surface with specimen or rea- gents used in testing. The stick is of a length sufficient to span the sides of a standard tray and has an extension at one end of accomodate an up¬ standing small projection at one side of the tray with the handle extending from the opposite side of the body of the stick over the opposite side of the tray. The body of the stick is notched at the end opposite the handle to accomodate spaced inwardly extending projections on the opposite side of the tray while the handle portion is rested thereon. The above and other features of the invention included various novel details of construction and combinations of parts will now be more particularly described with reference to the accompanying draw¬ ings and pointed out in the claims. It will be understood that the particular microtiter stick embodying the invention is shown by way of illustration only and not as limitation of the invention. The principles and features of this invention may be employed in varied and numerous embodiments without departing from the scope of the invention.
Brief Description of the Drawings
Figure 1 is a perspective view of a prior microtiter strip now used in performing immunoassay tests, and the like. Figure 2 is a similar perspective view of the present invention in the form of a stick that has shallow wells and a handle by the use of which the stick can be flipped to empty the shallow wells of their fluid contents at times while carrying out the immunoassay tests.
Figure 3 is a fragmentary perspective view of both the prior and present pieces assembled together in a tray used to support the pieces while carrying out the immunoassay tests. Figure 4 is a perspective view of the prior microtiter strip and the present stick with illu¬ stration being made of the manner in which their wells are filled from a squeeze bottle, the prior strip wells -having been individually filled while shallow wells of the present stick may be filled by flooding the top surface of the stick.
Figure 5 is a top plan view of the present welled test stick shown in actual size. Figure 6 is a longitudinal sectional view of the present stick as viewed on line 6-6 of Figure 5.
Figure 7 is a bottom plan view of the present stick. Figure 8 is an enlarged end view of the stick as viewed upon its handle end.
Figure 9 is an enlarged cross-sectional view taken on line 9-9 of Figure 5.
Figure 10 is an enlarged end view of the other end of the stick.
Figure 11 is an enlarged fragmentary longi¬ tudinal sectional view showing the interior of the wells, the raised boundary border, and the lensed well bottom portions as viewed also on line 6-6 of Figure 5.
Figure 12 is an enlarged fragmentary bottom plan view looking upon the bottom of the stick and into the lense bottom recesses.
Figure 13 is a perspective view illustrating the manner in which the well contents of the stick are dumped as by grasping the stick by its handle and with but a snap of the wrist dumping its con¬ tents into a sink drain rather than by using costly apparatus for decantation of the individual wells. Figure 14 is a perspective view of a photo¬ sensor arrangement for reading the residue showing upon the well lense portions at the completion of the tests. Detailed Description of the Invention
Allergies resulting from numerous contagions in the air have long been determined of individuals by bringing together samples of the individual blood with the various contagions pre-disposed respec¬ tively in a series of wells within immunoassay test strips constructed in one form or another. Such contagions can be categorized as pollens from trees, grass and weeds, mold spores, animal dander, bac- teria, dust mites and so on. The wells of the assay strips are pre-coated or supplied with the extracts of the contagion antigens prior to the carrying out of test with the blood specimens. When the blood sample of the patient is added and the excess blood subsequently decanted, the wells washed and with a tracer reagent, the wells will be colored in a degree to determine the extent that the patient is allergic to these contagions. With no coloring arising in any well, the patient will be regarded as immune to the particular allergy antigen of the one well.
Prior well strips 15 shown in Figures 1, 3 and 4 have been formed of transparent plastic and have a top flat strip 15' with a series of raised openings 16' from which depend cylindrical or test tube-like wells 16 that extend some one half inch in depth below the top flat strip 15'. The deep wells are interconnected with one another at their sides and their bottoms are closed by flat circular portions 16" upon which the residue results from the test will be observed. One end of the top flat strip 15* is extended at 17 and has a small hole 17' therein to fit over a small upstanding projection 19' of a tray 19 normally required for the support of the prior strips 15 to prevent spilling of their well contents. Figure 3. The other end of the top flat strip 15* is retained on the tray 19 at the opposite side between small inwardly-extending projections 19" that will lie astride end well portion 16 so that the well strip 15 is held against sliding displacement along the tray 19. A small window area 21 is provided in the opposite end extension 22 through which a raised indicia 23 on the tray frame may be read while the welled strip 15 over-lies the tray 19. A top hold down frame e 20 is placed over the collected strips 15 and such present sticks 25 which can be similarly assembled therewith upon this standard tray 19 to hold down all pieces against vertical displacement therefrom.
The prior deep welled strips 15 have to have their wells 16 filled individually with measured quantities of reagent or specimen as by the use of a pipette and that is a time consuming task. Decan- tation of the well 16 becomes a similar task and costly laboratory evacuation apparatus is required. Laboratory surroundings have had to be more or less available by the physician for the carrying out of immunoassay tests with these prior welled strips 15. These prior deep welled strips 15 can be placed on a flat supporting surface, but because of their height they cannot be placed with safety while having liquid contents in them. They are easily upset and thus are dependent upon the tray 20 for safe support. Thus, for filling and decantation -.' operations they need to be assembled between the tray parts 19 and 20.
The flat bottom plates 16" of the cylindrical wells 16 of the prior strip 15 make for reading of the test results as to color with the projection of light from through the well bottoms. Such flat bottom plates 16" make for the existence of often unclean circular corners against their cylindrical side walls and significant capillar effects arise thereby to reduce the accuracy of the readings.
The structural differences and advantages of the present microtiter stick 25 will now be made more apparent with the following detailed descrip- tion taken in connection with the drawing. Both the prior and present pieces are made of polystyrene, a plastic well known to have an affinity for capturing antigen and antibody proteins and to enhance the separation of solid matter during the solid phase cycles on pre-coating the stick wells and the performing of the immunoassay tests. The prior flat bottom well plate 16" of the prior strip 15 while having the same affinity for proteins takes on a heterogeneous coating preparation due to the diffi- culty in cleaning of the corners of the flat bottom plate 16" with its cylindrical wall and a hetero¬ geneous specimen assay will result.
Because of numerous deficiencies of the prior strip 15, the present stick has been designed to overcome such deficiencies to reduce labor in the use of the stick, facilitate the procedure of carrying out of immunoassay tests, lessen the costly laboratory equipment needed in the carrying out of the assay tests as with the prior microtiter strips 15, and finally to make the procedure more adaptable to physicians and laboratory personnel.
The present microtiter stick 25 principally differs in construction from the prior strip 15 by the provision of but shallow well depressions 26 fashioned in a solid plastic body that are bottomed with optically transparent concaved lense portion 26' that conform to the rounded well bottom. On projecting light upwardly through these lense portions 26" onto a photo-sensor 35 see Figure 14, the solid material on the lense portion will be focused on a plane and be more accurately read with the photo-sensor instead of merely sensing through an unfocused plate bottom 16" of prior strip 15. By the focused lighting arrangement a better color interpretation will be obtained. The present microtiter stick 25 is about the same size as the prior art strip 15 and can be placed therewith in the standard two-part try 19-20, see Figure 3. The present stick has an end lip 27 with a small hole 27' therein for assembly over the small upstanding projection 19' on the tray bottom part 19. The other end of the present welled stick 25 instead of a mere lip extension has an elongated handle extension 28 by which the stick 25 is handled at times of being placed upon the tray 19 or removed therefrom to empty its contents. This handle is also conveniently used to label the stick for patient identification purposes deemed critical for retaining the integrity of results. The top tray part 20 is cutaway at 20' on one side to accommodate the handle extension 28. To retain the handle end of the stick against lateral displacement the bottom of the stick is placed between two inwardly-extending projections 19" of the tray 19. The depending body of the stick has spaced vertically-extending re¬ cesses 29 and 30 under the handle extension 28 to respectively receive the small inwardly-extending frame projections 19" as the stick 25 is laid down upon the tray 19. It should be seen that the handle 28 can be easily grasped to lift the stick 25 from the tray 19 and this is often done many times during the assay to rid the shallow wells 26 of their contents. By the wells 26 being shallow decantation devices are unnecessary to empty the wells. Simply by a grasp of the handle 28 by the thumb and fore¬ finger the stick 25 can be lifted and with a snap of the wrist over a sink drain 31 in the manner illu¬ strated in Figure 13, the wells are quickly and completely ridded of their contents. All wells are at once emptied with the flip of the stick. Imple¬ ment decantation of the individual wells is there- fore not needed.
To ensure that complete emptying of wells is effected, cylindrical wall have been eliminated and the wells are less than full semi-spherical shape as they may appear in the sectional views. The arc for the well shape is struck from a center point 32 as seen in Figure 9 that is raised from the flush top surface of the stick, hence the wells at the top span a distance short of what would be the diameter for the center point, had not the center point been raised. A full diameter"would give hesitation to full dumping action and a dwell of the liquid over the top edge of the well. Thus, there is no sem¬ blance of a cylindrical well in the present stick.
An upstanding boundary border 31 surrounds all of the wells 26 running along the sides and across the ends of the stick to permit flooding of the top surface of the stick for effecting simultaneous filling of the wells 26 with unmeasured quantities as from a squeeze bottle as shown in Figure 4. Each well 26 holds 100 miσroliters, suitable in volume amount in drops for assay testing, and to obtain full colorization of the well surface. The resul¬ tant residue material will develop color and be read through bottom concaved lense area 26'. The boundary border 31 runs tangentially to the side edges of the wells 26 with no surface area lying therebetween. The border 31 runs to the full extent of the eight wells and is closed off at 31' on the end projection 27 and at 31" on the opposite handle end 28. The wells are fully confined by the boundary border 31 so that as illustrated in Figure 4 the entire series of wells 26 will by simple flooding with a squeeze bottle 32 or a free flowing device for spilling of the reagent or specimen on the top surface area, so that wells 26 will be instantly filled to measured capacity with little excess overflow, and each well holding 100 micro- liters. A stream of large specimen drops 34 will abound from bottle nozzle 33 to flood the top surface of the stick 25 and instantly fill the wells 26. It should be apparent that this flooding procedure cannot be effected with the deep welled prior stick 15 and as illustrated in Figure 4 the deep cylindrical wells will have had to have been filled separately and individually with the drops 34 from the squeeze bottle 32. Any measured quantity of blood specimen could have been ascertained only by counting the drops 34, short of the actual filling of the wells 16, unless and more often a pipette would have been used to discharge measured quantities into cylindrical wells.
It should be further apparent that with prior strips 15 comprising the deep transparent cylin¬ drical test tube-like wells lying attached and closed upon one another that the optical readings of one well can be hampered by optical interference of the showing of the other wells so that true readings of the respective wells are not so likely to pre¬ vail. With the present stick 25, the construction is inherently different. The wells 26 are not tubes as with the prior stick 15, but are mere shallow and frosted openings formed with inclinded walls in the stick's solid body and without cylindrical well walls and such that there can be little, if any, optical interference between adjacent wells.
To further assure against possible optical interference, the present stick 25 is frosted throughout all of its surface areas except for the relatively small surface area of the well lensed portions 26'. The frosting extends over all sur¬ faces, top, sides, bottom and down into the shallow wells 26, except for their lensed bottoms 26'. However, it can be sufficient if but the wells and top surface can be frosted. Any light transfer from a source, not only between wells, will be diffused by the frosting. The wells by themselves will also be given by the frosting therein effective added contact surface area to ensure that adequate coating densities will be obtained for better observation of test results. The affinity action of proteins upon the polystyrene thus has been enhanced by the frosting.
The frosting will have first been made upon the molding surface of the stainless steel die parts and such frosting transferred during the molding oper¬ ation onto the molded work body. The die parts are frosted by an electroylytic etching procedure or by any other accepted frosting procedures. Corres- ponding frosting upon the work stick is thus ob¬ tained from the dies. To ensure that the lense portions 26' will be clear, the lense areas of the dies are smoothly ground toward infinite fineness and its clarity is transferred to the plastic work stick body. From so formed die parts, sticks frosted except from the lense areas are produced ready for use, inexpensively and are disposable • after final use in the carrying out of the assay tests. The sticks will have been pre-coated with specified antigen or antibody proteins prior to their delivery to physicians and laboratories, and ready for use.
The present stick 25 has a flat bottom surface in which the area beneath the lense portions 26 are recessed. Such recesses will keep the lense por¬ tions from being scratched. The flat bottom surface allows the present stick to be rested upon a flat supporting surface and not easily turned over, thereby permitting its use without the need for tray 19. With the frosting extending over the handle data marks can be made thereon.
The present stick will be pre-coated with antigens and antibodies in accordance with the physician's specifications and requirements and varying for different geographical locations. There are different allergy contagions for the different locations which will be predetermined and known to the physician within his given geographical area. Some of the common allergy contagions are pollen from trees, grass and weeds, mold spores suspended in air, on surface and in bread, animal dander, bacteria, dust mites, and so on. The wells of the present stick are pre-coated with any of these antigens, one antigen for each well and such other antigens not named that are available in serum form for the purpose. In the pre-coating of these sticks, all wells can be filled simultaneously in measured quantities of the different antigens by a multiple dispensing apparatus upon advancing the sticks thereunder in a step by step procedure.
After the stick wells have been dehydrated and the antigens reacted with the polystyrene surface of the wells, the sticks are made ready for distribution to physicians and laboratory users.
In the use of these sticks, the physicians and users will merely supply the patient's specimen and reagents and wash liquids to carry on known steps of procedure until a color reading will be seen of more or less color density of the residue material overlying the frosted well surfaces and on the unfrosted lense areas 26'. Ordinarily, the results of the assay test can be readily determined by merely viewing the well for the color intensities but for a roe accurate determination by subjecting the stick to a light source 37, Figure 14, and at the proper focal point the residue upon the lense may be observed with a photo-sensor 35. All the readings will be free of optical interferences between wells due to the wells being shallow and frosted. In the carrying out of these tests, usual decantation of the individual wells has been unneces¬ sary. The physician will have emptied the well contents by grasping of the handle and with a mere snapping of the wrist disposing of the well contents into a sink drain. The procedure requires reagents, wash liquids and color developing liquids as well as the blood specimens, the use of all of which will be greatly expedited with the present stick. Once the allergy has been determined, its antigens can be injected into the patient to give him immunity and freedom from his ailment. It should be understood the present stick can be used for tests other than for blood allergies as for urine and other natural fluids. With the sticks being pre-coated with reagents used in carrying out such other clinical tests and need only the addition of the natural specimen to determine the nature of possible ailment of the patient. As well as immuno¬ assay tests, micro, solid phase enzyme-linked adsorbent assays (micro ELISA) can be carried out with the present sticks. The veterinarian will also find these sticks handy in carrying out tests with animal fluids.
It is understood that both antigens and anti¬ bodies are proteins and that they lock with one another upon being brought together in the assay tests. Each of the wells of the microtiter stick is coated with but a specific antigen or antibody depending upon the nature of the assay to be made. Because of the natural affinity of these proteins to a hydropholic surface such as that of polystyrene, these two proteins irreversibly bind, absorb or combined with the pre-coated surface upon the well walls. By frosting the well wall surface enlarges the availability surface of the well and thereby the capacity and the efficiency of the well surface is increased. Excess solution of the antigens or antibodies are decanted and washed free of the well matrices.- the well matrix is air dried and stored for use in determining any of the many possible conditions of the patient. The physician simply adds the blood specimen of the patient to the coated well stick. After incu¬ bation any corresponding antibody in the specimen will bind or lock with corresponding antigen (aller- gen) of the antigen-coated well. The quantity of the antibody so bound depends upon how much there is of the antibody in the body fluid, the amount of the antibody that has been captured on the well wall surface, the affinity of the capture, the proximity of contact between the entire body of the specimen and the coated surface, the homogenity of the exposure of specimen body to the coated well wall and the spacial molecular arrangement of the anti¬ gens. Once the antigen pre-coated wells have been exposed to the human blood specimen, and the corres¬ ponding antibodies bound and locked with the pre- coated antigens of the wells, the wells are incu¬ bated. After suitable washing and removal of unbound material suitable animal antibodies having enzymes and have color forming molecules attached to them are added to the well already pretreated with the human blood specimen, and with further incu¬ bation, washing and removal of material and then by the use of subsequent corresponding enzyme sub¬ strate, the well surfaces will develop color of intensities proportional to the level of antibodies contained in the patient's blood specimen and with the degree of coloring being noted, the physician is immediately advised of the type and degree of the allergy of his patient. The assay will have been finally completed.
The degree or intensity of the color is propor¬ tional to the amount of antibody that was bound upon the well wall from the body specimen. These proce¬ dures have been very successful for measuring different types of antibodies and will be greatly enhanced and improved when carried out with the present sticks. Costly special filling and decari- tation equipment will no longer be necessary. Deep wells are no longer a problem. By the provision of the bottom well lense, the interpretation of the color development has been greatly facilitated.
The prior microtiter strip 15 and its bottom plate 16" has had many optical shortcomings since it has only provided low plate surface area for the specimen and cylindrical wall areas of the well optically finished and not involved in optical observation. Color development has been less easily delineated due to inter-well interference due to such optically clear surfaces but unavailable for the viewing or sensing color development. With the present shallow walls the full surface areas are available for sensing of color development. Further, it should be seen that the present stick has been designed for easy molding. That the present stick 25 is less than one-half the height of the prior strip 15 and can be safely rested upright on a flat supporting surface without the need of a tray and at the same time can be used in the trays along with the prior strips. While various changes may be made in the details construction, it shall be understood that such changes shall be within the spirit and scope of the present invention as defined by the appended claims.

Claims

1. A microtiter stick adapted for use in per¬ forming immunoassay tests comprising a solid plastic body having one or more shallow wells therein upon surfaces of which resultant residue may be collected from the tests and observed, each well being rounded on its bottom and struck from a compass point externally of its internal space and the diameter of the opening being of less diameter that of a circle that would be prescribed therefrom.
2. A microtiter stick adapted for use in per¬ forming immunoassay tests as defined in claim 1 and said plastic body being elongated and the shallow well openings running in series there- along and a handle portion projecting from one end of the body by which the stick may be grasped and flipped to rid the wells of their contents.
3. A microtiter stick adapted for use in per¬ forming immunoassay tests as defined in claim 1 and the bottom of said wells being in the form of a concave lense portion through which an image of residue extending over its surface area may be projected from the plastic body upon a micro-sensor or screen.
4. A microtiter stick adapted for use in per¬ forming immunoassay tests as defined in claim 3 and said plastic body having a flat bottom supporting surface and being recessed to expose the lense portion of the well removed from the flat bottom surface whereby to protect the lense portion against being scratched upon the stick being placed upon a supporting surface.
5. A microtiter stick adapted for use in per- forming immunoassay tests as defined in claim 2 and said plastic body having a top flat surface about the well opening and a boundary border about the flat surface and the well openings whereby the wells may be filled by controlled flooding of the top surface about the well opening with specimen or reagents used in the test.
6. A microtiter stick adapted for use in per¬ forming immunoassay tests and the like as defined in claim 2 and said elongated plastic body being of a length to span the sides of a standard tray and having an extension at one end to accomodate the upstanding small pro¬ jection at one side of the tray and said handle portion extending from the opposite end of body portion over the other side of the tray, said body portion being notched at the opposite sides of its handle end to accomodate spaσed inwardly-extending projections on the opposite side of the tray while the handle portion is rested thereover.
7. A microtiter stick adapted for use in per- forming immunoassay tests and the like as defined in claim 3 and the surfaces of the wells being frosted except for the bottom lense area to allow added residue build upon the major well surface and reduce optical color interference between adjacent wells within the body.
8. A microtiter stick adapted for use in per¬ forming immunoassay tests as defined in claim 6 and 7 the plastic body being further frosted through its other surfaces therefor and over the handle portion to permit data information to be marked thereupon and to improve the grip when used.
EP87900414A 1985-11-27 1986-11-24 A disposable microtiter stick Ceased EP0248882A1 (en)

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US5098661A (en) * 1988-11-16 1992-03-24 Medical Laboratory Automation, Inc. Coded cuvette for use in testing apparatus
CA2041050A1 (en) * 1990-10-29 1992-04-30 Kerinchan Babaoglu Batch test apparatus with adjacent negative control and patient sample rows
US6027695A (en) * 1998-04-01 2000-02-22 Dupont Pharmaceuticals Company Apparatus for holding small volumes of liquids
ES1042388Y (en) * 1998-11-03 2000-02-01 Grifols Grupo Sa ALVEOLAR SUPPORT FOR ANALYSIS SAMPLES.
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GB2428794A (en) * 2005-08-02 2007-02-07 Advanced Biotech Ltd Two part microwell plate and method of fabricating same
GB201322082D0 (en) * 2013-12-13 2014-01-29 Ge Healthcare Bio Sciences Ab Floatable microplate

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WO1987003218A1 (en) 1987-06-04
IL80763A0 (en) 1987-02-27
AU6773287A (en) 1987-07-01
CA1289871C (en) 1991-10-01

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