Apparatus for blot screening numerous, small volume, antibody solutions

• US 4,834,946 A
• Filed: 02/05/1987
• Issued: 05/30/1989
• Est. Priority Date: 02/05/1987
• Status: Expired due to Term

• Current Assignee: Andrew E. Levin
• Original Assignee: Andrew E. Levin
• Inventors: Levin, Andrew E.
• Primary Examiner(s): Richman, Barry S.
• Assistant Examiner(s): WARDEN, JILL ALICE
• Application Number:
  US07/011,291
• Time in Patent Office: 845 Days
• Field of Search: 422/100, 422/101, 435/285, 435/288, 435/292, 435/293, 435/301
• US Class Current: 435/287.2
• CPC Class Codes: B01J 2219/0043 for direct application of reagents, e.g. through openings in a shutter
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/5302 Apparatus specially adapted for immunological test procedures
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

A cover plate, having a lower, flat surface containing an array of numerous, parallel, closely spaced apart, downwardly opening channels, is releasably clamped upon a flat support plate surface. The channels each have inlet and outlet holes communicating with the upper surface of the cover plate. A paper-like, resilient cushion sheet is clamped between the plate surfaces and a paper-like membrane, carrying a pattern of antigen-like material, is arranged between the sheet and the cover plate channeled surface. The channels are very narrow, shallow and many times longer than their widths and are closed by the stripe-like portions of the membrane which overlie the channels. The cushion sheet is compressed between adjacent channels and bulges upwardly at each of the channels to thereby bulge the membrane stripe-like portions into dome-shaped cross-sections extending into the channels. This reduces the volumes of the channels and increases the surface areas of the membrane stripe-like portions that are exposed within the channels. Numerous drop-size samples of antibody-type solutions are inserted into the channels through channel inlet holes, that extend through the cover plate, for reacting with the material carried on the stripe-like portions of the membrane. A removable manifold having a pair of elongated grooves may be temporarily applied upon the cover plate so that the grooves overlie all of the channel inlet holes and all of the channel outlet holes for rapidly, simultaneously filling and draining all the channels.

Claims

1. An apparatus for use in blot screening antibody-type material by simultaneously reacting a substantial number of very small volume liquid samples of such material arranged in a stripe-like pattern with a transversely arranged antigen pattern carried upon a paper-like membrane, comprising:

a base plate having an upper, flat support surface, and a cover plate having a lower, flat cover surface overlapping the base plate support surface;

manually releasable fasteners clamping the plate overlapping surfaces together;

a thin, resilient cushion support sheet positioned between and substantially covering the plate surfaces in face-to-face contact with the base plate support surface;

an array of a substantial number of parallel, closely spaced, elongated, shallow and very narrow, downwardly opening channels formed in the cover plate support surface separated by stripe-like portions;

each channel having at least one small filling hole extending through the cover plate for conveying a sample of a liquid material into the channel;

a membrane removably positioned upon said cushion sheet and arranged in face-to-face contact with the cover plate surface so that the membrane closes the channel openings, said membrane being adapted for screening samples of liquid materials;

and said resilient support sheet being resiliently compressed against the base plate support surface between the channels by the stripe-like portions of the cover plate surface that are located between the adjacent pair of channels and the cushion support sheet resiliently bulging upwardly at the channel openings and thereby bulging the stripe-like portions of the membrane into the channels for forming inwardly extending, dome-like in cross-section, bottom walls for the channels which walls simultaneously reduce the interior volumes of the channels and increase the membrane surface area exposed to liquids contained within the channels;

wherein a sample of a liquid material may be introduced within at least one of the channels and another sample of a different liquid material may be introduced within at least one of the other channels, with the portions of the membrane surface area being bulged by the resilient support sheet into the channels to close the bottom of each respective channel, reduce the volume of each respective channel by extending into each respective channel and substantially prevent leakage of sample between adjacent channels.

2. An apparatus as defined in claim 1, and further including each channel having a second, small hole extending through the cover plate communicating therewith, with the filling hole of the channels being aligned in a row near one end of the channels and the second holes being aligned in a row near the other end of the channels wherein liquid may be flowed into the channels through one row of filling holes and flowed out of the channels through another row of second holes.

3. An apparatus as defined in claim 1, wherein the width of said channels is in the range of 1.5-5 millimeters and the length of said channels is greater than 20 times such width, so that the channels may be filled with microliter, drop-size samples;

and with the distance between pairs of adjacent, parallel channels being approximately equal to the width of said channels.

4. An apparatus as defined in claim 3, wherein the resilient cushion support sheet is formed of a foam type of plastic material which is characterized by being relatively inert to the specimen materials.

5. An apparatus as defined in claim 1, and further including a manifold block releasably secured upon the cover plate and having a downwardly opening groove which extends over the filling holes, and an opening extending through the block communicating the groove with the upper end of the block, so that a liquid may be flowed through the opening, the groove and the cover plate filling holes and into a number of channels simultaneously.

6. An apparatus as defined in claim 1, and including a second array of parallel channels, similar to the first-mentioned array of channels, located in the lower, flat cover surface of the cover plate adjacent to the first-mentioned array of channels and having similar filling holes extending through the cover-plate, wherein a second cushion support sheet, similar to the firstmentioned sheet, is arranged between the base plate, and cover plate and an additional membrane, is located between the resilient sheet and the base plate, at which the second array is formed, whereby two separate arrays may be simultaneously used for screening.

7. An apparatus as defined in claim 1, wherein said membrane includes a nitrocellulose material.

8. An apparatus as defined in claim 7, wherein said resilient support sheet includes a polyethylene material.

9. An apparatus as defined in claim 1, and wherein said filling holes are aligned in a row near one end of the channels, and further comprising a row of second holes communicating each channel to the upper surface of the cover plate and the second holes being aligned in a row;

a manifold block releasably secured upon the cover plate and having a pair of spaced apart, downwardly opening grooves, with one of said grooves communicating with said row of filling holes and the other of said grooves communicating with said row of second holes;

each groove having an opening extending through the manifold block and communicating the respective groove to the upper end of the block;

whereby liquid may be flowed through one opening and its respective groove and said filling holes into all of the channels and the row of second holes permit the liquid to flow out of the channels so that liquid may be circulated into and out of all of the channels simultaneously.

10. An apparatus as defined in claim 9, wherein said manifold block is generally U-shaped in cross-section, formed with an integral base and legs, and being of a length and width sufficient to overlap all of the parallel channels in the cover plate, and with one of said grooves being located in the bottom of each of the said legs forming the U-shaped block.

11. An apparatus as defined in claim 10, and including a rigid bar extending across the top of the cover plate transversely to the parallel channels and releasably secured to the cover plate by a fastener;

and a second fastener securing the base part of the U-shaped block to the bar with the bottoms of the legs of the block, which contain the downwardly opening grooves covering the rows of filling and second holes, being clamped against the upper surface of the cover plate.

12. An apparatus as defined in claim 9, wherein said manifold block is formed of a transparent, plastic material, and having a lower surface arranged to engage the upper surface of the cover plate in face-to-face contact therewith, and said manifold grooves being formed in the manifold block lower surface, and with the peripheral edge portions of the grooves being releasably sealed against the cover plate upper surface.

13. An apparatus for use in blot screening solutions of antibody and the like materials by simultaneously, separately reacting a substantial number of microliter-size volume samples of such material with an antigen-like pattern carried upon a paper-like membrane, comprising:

a base plate having a flat support surface and a cover plate having a lower, flat, cover surface overlapping the base plate support surface;

manually operable screws extending through the cover plate into threaded openings in the base plate for manually releasably clamping the two plates together;

an array of a substantial number of parallel, closely spaced, elongated, shallow, very narrow, downwardly opening channels formed in the cover plate lower surface separated by stripe-like portions;

means defining holes extending through the cover plate for communicating each channel with the upper portion of the cover plate for conveying liquid specimens into and out of the channels;

a thin, resilient, spongy cushion sheet positioned upon the base plate support surface beneath the channels, and a membrane removably positioned upon the cushion sheet and in face-to-face contact with the cover plate lower surface covering each of the downwardly extending openings of the channels;

said membrane being adapted for screening samples of liquid materials;

said cushion sheet being of a relatively low density so that it is characterized by having sufficient resiliency to be compressed between the cover plate areas located between adjacent channels, while resiliently bulging towards the channel openings so as to bulge the membrane into the channels considerably reducing the interior volume of the channels while increasing the membrane surface area which is exposed to liquids contained within each channel;

wherein a sample of a liquid material may be introduced within at least one of the channels and another sample of a different material may be introduced within at least one of the other channels, with the membrane located at the channel openings bulged by the cushion sheet and exposed to the samples, whereby leakage between adjacent channels is substantially prevented.

14. An apparatus as defined in claim 13, wherein the holes into each channel are arranged in two rows, one row near one end of the channels and the other row near the opposite end of the channels;

and further including a manifold block having a lower surface releasably engaged with the cover plate at an upper surface thereof and having downwardly opening grooves overlapping the two rows of holes, with each groove communicating through a passageway to the lower surface of the block so that liquid may be inserted through one groove into all of the channels simultaneously through one row of holes and removed from all of the channels through the opposite row of holes;

and further including releasable fastening means securing the manifold block in engagement to the upper surface of the cover plate so that the block may be applied upon or removed from the cover plate while the cover plate remains clamped to the base plate by the manually operable screws.

15. An apparatus as defined in claim 14, and including a resilient sealing ring arranged around each of said manifold block grooves and sealing against the adjacent cover plate portions around the respective rows of holes.

16. An apparatus as defined in claim 14, and said manifold block being formed of a transparent, plastic material so that the grooves and passageways may be visually observed during use thereof.

17. An apparatus as defined in claim 14, wherein said manifold block releasable fastening means comprises manually operable screws which extend through and fasten the block, the cover plate and base plate together, and which can be manually released, for removing the block, and replaced for fastening the base plate and cover plate together without the manifold block.

18. An apparatus as defined in claim 13, wherein said membrane includes a nitrocellulose material.

19. An apparatus as defined in claim 18, wherein said cushion sheet includes a polyethylene material.