Search Images Maps Play YouTube News Gmail Drive More »
Sign in
Screen reader users: click this link for accessible mode. Accessible mode has the same essential features but works better with your reader.

Patents

  1. Advanced Patent Search
Publication numberUS3883398 A
Publication typeGrant
Publication date13 May 1975
Filing date7 May 1973
Priority date7 May 1973
Publication numberUS 3883398 A, US 3883398A, US-A-3883398, US3883398 A, US3883398A
InventorsKazuyuki Ray Ono
Original AssigneeBellco Glass Inc
Export CitationBiBTeX, EndNote, RefMan
External Links: USPTO, USPTO Assignment, Espacenet
Microculture slide chamber
US 3883398 A
Abstract
A microculture slide chamber for simultaneously growing a plurality of mono-layer cell cultures on a slide includes a matrix which defines a plurality of generally parallel holes extending therethrough so that a plurality of wells for containing the culture bearing fluid are provided when the matrix is in abutment with a slide. Surrounding each hole on one side of the matrix is a gasket that is integral with the matrix and serves to seal the fluid in each well. A pressure plate with mating holes overlies the opposite side of each matrix and spring clips hold the plate, matrix and slide in assembled relation. A cover for the microculture slide chamber is provided.
Images(1)
Previous page
Next page
Description  (OCR text may contain errors)

United States Patent [1 1 Ono [ May 13, 1975 MICROCULTURE SLIDE CHAMBER [75] Inventor: Kazuyuki Ray Ono, Bridgeton, NJ. [73] Assignee: Bellco Glass, lnc., Vineland, NJ. [22] Filed: May 7, 1973 [21] Appl. No.: 357,991

[52] US. Cl 195/127; 195/139 [51] Int. Cl Cl2b 1/00 [58] Field of Search l95/l39, I27

[56] References Cited UNITED STATES PATENTS 3,745.09] 7/1973 McCormick 195/139 3,791,933 2/1974 Moyer ct al. [95/127 Primary Examiner-Alvin E. Tanenholtz Attorney, Agent, or FirmSeidel, Gonda & Goldhammer [57] ABSTRACT A microculture slide chamber for simultaneously growing a plurality of mono-layer cell cultures on a slide includes a matrix which defines a plurality of generally parallel holes extending therethrough so that a plurality of wells for containing the culture bearing fluid are provided when the matrix is in abutment with a slide. Surrounding each hole on one side of the matrix is a gasket that is integral with the matrix and serves to seal the fluid in each well. A pressure plate with mating holes overlies the opposite side of each matrix and spring clips hold the plate, matrix and slide in assembled relation. A cover for the microculture slide chamber is provided.

10 Claims, 2 Drawing Figures MICROCULTURE SLIDE CHAMBER This invention relates to a microculture slide chamber. More particularly, this invention relates to a microculture slide chamber for simultaneously growing a plurality of mono-layer cell cultures on a slide or similar structure.

There are many types of laboratory tests, particularly in the biological sciences, wherein it is desirable to form cell cultures on a slide. Among these are monolayer cell cultures for which the present invention is particularly suited, although those skilled in the art will recognize that it may have other uses. For various reasons, it is desirable that two or more cell cultures be grown on the same slide. For example, an advantage of growing multiple cell cultures on the same slide is that different viruses can be used to innoculate the same type of cells or, in the alternative, one virus can be used to innoculate different types of cell cultures. There are, of course, other advantages as hereinafter described. The present invention provides a microculture slide chamber which enables such persons to gain these advantages.

Prior attempts have been made to provide microculture slide chambers for growing a plurality of cell cultures on the same slide. Among these is a tissue culture chamber-slide sold by Miles Laboratory, Inc. under Lab-Tek Catalog No. D2270. This device consists of a glass slide with a removable non-toxic gasket and an attached optically clear plastic sample chamber. The chamber-slide has proven to be quite unsatisfactory for several reasons. Among these is that the fluid occasionally leaks from chamber to chamber. Worse still, the separation of the slide from the remaining parts of the device requires the application of pressure to break the gasket seal followed by the removal of the gasket with forcep. In use, the breaking action or the removal of the gasket results in the removal of parts of the culture cells, a wholly unacceptable result.

The present invention overcomes the disadvantages of such prior art devices by providing a readily assemblable and disassemblable microculture slide chamber for simultaneously forming two or more culture cells or the like on a slide. Such cell cultures may be, but need not be, mono-layer cell cultures.

The microculture slide chamber comprising the present invention is designed so that it will have the advantages of consistency from slide to slide, convenience in use, be economical to purchase and maintain, and can be readily stored. The advantage of forming a plurality of cell cultures on a single slide is that there can be consistency in cell staining, consistent histochemistry, and identical manipulation. Such a device is convenient because it provides efficiency in processing and examination of all cells on a single slide and also allows for storing the cells in racks that can be readily identified. Economy is achieved because the microculture slide chamber is recyclable except, of course, for the slide itself.

In accordance with the present invention, the microculture slide chamber comprises a slide on which the cultures are to be grown together with a matrix made of resilient material through which extend a plurality of holes. The matrix is positioned in abutting relation to the slide so that each hole defines a well or chamber within which the culture bearing fluid can be maintained. The side of the matrix which abuts the slide includes gaskets integral therewith and surrounding each of the holes. The gaskets provide a fluid tight seal for each well thus formed preventing leakage from well to well. Overlying the opposite side of the hole is a plate serving as a pressure plate. The plate has a plurality of holes extending therethrough which holes are arranged on the plate so as to align with the matrix holes. A pair of spring clips engage the side edges of the surface of the slide and plate to retain the same in abutting relation with the matrix. A cover is provided for closing the wells while the cell cultures are being formed.

A microculture slide chamber constructed in accordance with what is described and claimed herein has many uses and applications. Among these are the following:

Screening for effects of compounds on cellular morphology, replication, viability and differentiation.

2. Study of the effects of compounds on the cellular uptake of a variety of radio isotopic substrates. This application requires the use of a polymer substrate or slide.

3. Autoradiographic studies permit the localization of radio isotopic tracers in the cells.

4. Viral titrations can be rapidly and easily performed in the chambers.

5. Cytotoxic antibody titrations can be done as well.

6. Lymphocytotoxic assays using either optical or radio-isotopic end points are easily performed.

7. lmmunoflourescence methods are especially easy with this invention. The detection of anti-nuclear antibodies (ANA) for testing for lupus erythematosis is very easy. Viral diagnosis could be accomplished in this system by titrating for immunoflourescence of the patients serum against preinfected cell mono-layers.

8. It is theoretically possible to perform the macrophage inhibition factor assay using this invention.

For the purpose of illustrating the invention, there are shown in the drawings forms which are presently preferred; it being understood, however, that this invention is not limited to the precise arrangements and instrumentalities shown.

FIG. 1 is an exploded perspective view of a microculture slide chamber in accordance with the present invention.

FIG. 2 is a transverse sectional view of the microculture slide chamber showing the same in assembled relation.

Referring now to the drawings in detail, wherein like numerals indicate like elements, there is shown a microculture slide chamber designated generally as 10.

The microculture slide chamber 10 is provided with a slide 12 which may be made of glass, as is conventional. The slide 12 is somewhat longer than the microculture slide chamber 10 so that a frosted portion 13 upon which identifying marks can be made protrudes outwarding from the cell. Overlying the slide 12 is a matrix 14 best shown in FIG. 1. The matrix 14 is made of a resilient material such as silicone rubber. Of course, other resilient materials may be substituted provided that they are autoclavable. Extending through the matrix 14 are a plurality of holes 16 in generally parallel relation to each other. Although the matrix 14 may take any form, such as circular or even toroidal, it is shown as being generally in the shape of a rectangular parallel-piped having opposed planar surfaces 18 and 20 between which the holes 16 extend. Ten holes 16 are shown extending from surface 18 to surface 20.

However, any number of such holes, as desired, may be provided.

When matrix 14 is brought into abutting relation with the upper surface of slide 12, each of the holes 16 defines a well or chamber for containing the culture bearing fluid. To be certain that the fluid does not leak from well to well, a gasket 22 surrounds each of the holes 16. The gaskets 22 are integrally formed with the matrix 14 and provide a fluid tight seal when the slide 12 is held in abutting relation with the matrix 14 under the force of the spring clips 24 and 26. The advantage of making the gaskets 22 integral with the matrix 14 is that they are simultaneously removed when the matrix is separated from the slide, thus avoiding the danger that their individual removal with forceps would result in damage to the cell cultures.

Overlying the matrix 14 is a pressure plate 28 whose function is to cooperate with the clips 24 and 26 to apply a compression force on the matrix 14 to retain the gaskets 22 in good contact with the slide 12. Plate 28 is preferably made of stainless steel or an autoclavable polymer such as polycarbonate. The plate 28 is dimensioned to have the same width as the matrix 14 just as the matrix 14 has a width approximately equal to the width of the slide 12. Moreover, the plate 28 is provided with holes 30 extending therethrough which holes are positioned in the plate 28 so as to align with the holes 16 in the matrix 14. Thus, the holes 30 permit ready access to the holes 16 and the wells thus defined when the microculture slide chamber is in its assembled condition. If desired, plate 28 may be permanently fixed to matrix 14.

Clips 24 and 26 provide a resilient means for main taining the microculture slide chamber in its assembled relation. As shown, each of the clips 24 and 26 is made of a tempered spring metal, such as stainless steel, and it is provided with curved dependent edge engaging sections 32 and 34. The edge engaging sections 32 and 34 depend from a medially curved or bent intermediate section such that the distance between the edge sections 32 and 34 is normally less than the thickness of the assembled microculture slide chamber including the slide 12, matrix 14 and plate 28 when each clip is in an unstressed condition. Each of the clips 24 and 26 slides over the edges of the slide 12 and the plate 28 and thereby compresses them toward the matrix 14, thus retaining the microculture slide chamber in its assembled relation without interferring with access to the holes 30. A cover 36 is made of polycarbonate polymer or some other autoclavable material and preferably is light transparent so that the interior of the hole 16 can be observed. Cover 36 is dimensioned to overlie the entire surface of plate 28 and, of course, the matrix 16 thereby protecting the culture fluid when placed in the holes 16.

It should be understood that the number of holes 16 in matrix 14 can be varied as desired. Moreover, the dimensions can also be varied. in one preferred embodiment, the holes 16 are sized to contain culture fluid of 0.4 ml maximum with a recommended amount of approximately O.2 ml. The slide 12 is preferably of stan dard dimensions for microscope slides which are approximately 09 mm to 1.09 mm in thickness.

In use, the microculture slide chamber is assembled and the appropriate culture bearing fluids are placed in two or more of the holes 16. The cultures are innoculated, the cover 36 is placed on top of the microculture slide chamber, and the entire device is placed in a CO incubator. As is known, the cells attach to the base of the glass slide 12 to form a monolayer cell culture at the bottom of each of the wells defined by the combination of the slide 12 and the holes 16 in the matrix 14. The cells can be innoculated with viruses, or any of the procedures outlined above can be effected.

Once the monolayer culture has been formed, it is a straightforward procedure to disassemble the microculture slide chamber, fix and stain the cells, thereafter examine them under an appropriate microscope. Cover slips can also be fastened to the cells to make a permanent record of the culture.

The use of the invention is not limited to glass microscope slides. Other types of slides, such as plastic slides to which the cells will adhere, can be used. One such plastic is a polyester sold under the trade name Melinex" and manufactured by Imperial Chemical Industries, Inc. The advantage of using a plastic is that it can be readily divided, if desired after formation of the cells. This may be advantageous when it is desirable to measure the radioactivity of each cell monolayer.

The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof and, accordingly, reference should be made to the appended claims, rather than to the foregoing specification as indicating the scope of the invention.

I claim:

1. A microculture slide chamber for growing cultures comprising:

a slide on which said cultures are to be grown;

a matrix having a plurality of generally parallel matrix holes extending therethrough, said matrix being made of a resilient material;

said matrix holes when positioned over said slide defining a plurality of wells for containing a culture bearing fluid;

a gasket surrounding the end of each matrix hole adjacent said slide, each said gasket being integral with said matrix, and each said gasket providing a fluid-tight seal between said slide and said matrix for retaining fluid in said wells;

a plate having openings therethrough aligned to permit access to said matrix holes when said plate is in overlying relation to said matrix;

resilient clip means for retaining said microculture slide chamber in assembled relation for growing cultures therein, said assembled relation comprising said slide in abutting relation with the gasket side of said matrix and said plate in abutting and aligned relation with the opposite side of said matrix, and

a cover for said microculture slide chamber.

2. A microculture slide chamber in accordance with claim 1 wherein said clip means comprise first and sec ond resilient clips for engaging the edges of said slide and plate and Compressing them into engagement with said matrix.

3. A microculture slide chamber in accordance with claim 1 wherein said matrix is made of a silicone polymer.

4. A microculture slide chamber in accordance with claim 1 wherein said matrix, said clip means. said plate and said cover are made of autoclavable substances.

5. A microculture slide chamber in accordance with claim 4 wherein said cover is made of a polycarbonate polymer.

6. A microculture slide chamber in accordance with claim 4 wherein said clip means is made of a stainless steel.

7. A microculture slide chamber in accordance with claim 4 wherein said plate is made of stainless steel.

8. A microculture slide chamber in accordance with claim 1 wherein said plate is permanently fixed to said matrix.

9. A microculture slide chamber in accordance with claim 1 wherein said slide is made of a polymer material.

10. A microculture slide chamber for growing cultures on a slide comprising:

a matrix having a plurality of generally parallel matrix holes extending therethrough, said matrix being made of a resilient material;

said matrix holes when positioned over a slide defining a plurality of chambers for containing a culture bearing fluid;

a gasket surrounding one end of each matrix hole on one side of said matrix, each said gasket being integral with said matrix, and each said gasket being capable of providing a fluid-tight seal between a slide and said matrix for retaining fluid in said chambers when a slide is placed against the gasket side of the matrix;

a pressure plate having openings therethrough aligned to permit access to said matrix holes when said plate is in overlying relation to said matrix; and

resilient clip means for retaining said microculture slide chamber in assembled relation with a slide for growing cultures on the slide, said assembled rela tion comprising a slide in abutting relation with the gasket side of said matrix and said plate in abutting and aligned relation with the opposite side of said matrix.

Patent Citations
Cited PatentFiling datePublication dateApplicantTitle
US3745091 *18 Nov 197010 Jul 1973Miles LabBiological reaction chamber apparatus
US3791933 *25 Feb 197112 Feb 1974GeometRapid methods for assay of enzyme substrates and metabolites
Referenced by
Citing PatentFiling datePublication dateApplicantTitle
US4039247 *12 Dec 19752 Aug 1977National Research Development CorporationDevice for use in testing of fluid samples on microscope slides
US4284725 *13 Aug 197618 Aug 1981Dynasciences CorporationVirus titration and identification system
US4396717 *21 Jul 19812 Aug 1983Biotest-Serum-Institut GmbhNutrient medium carrier system
US4728607 *22 Mar 19841 Mar 1988J. K. And Susie L. Wadley Research Institute And Blood BankMiniaturized yeast identification system
US4847128 *14 Jul 198611 Jul 1989Wadley Technologies, Inc.Miniaturized yeast identification system
US5081033 *14 Jul 198614 Jan 1992Wadley Technologies, Inc.Miniaturized yeast identification system
US5192503 *23 May 19909 Mar 1993Mcgrath Charles MProbe clip in situ assay apparatus
US5328843 *26 Feb 199112 Jul 1994Jun FukudaMethod for allocating cells and cell allocation device
US5364790 *16 Feb 199315 Nov 1994The Perkin-Elmer CorporationIn situ PCR amplification system
US5484731 *26 May 199316 Jan 1996Becton, Dickinson And CompanyMultiwell in-vitro fertilization plate
US5527510 *5 Jun 199518 Jun 1996The Perkin-Elmer CorporationIn situ PCR amplification system
US5552321 *24 Aug 19933 Sep 1996Bioptechs Inc.Temperature controlled culture dish apparatus
US5571721 *5 May 19945 Nov 1996Erie Scientific CompanyImproved biological culture slide and method of making same
US5619428 *31 May 19958 Apr 1997Neopath, Inc.Method and apparatus for integrating an automated system to a laboratory
US5675700 *5 Jun 19957 Oct 1997The Perkin-Elmer CorporationAssembly system for in situ per amplification
US5681741 *8 Sep 199528 Oct 1997The Perkin-Elmer CorporationIn situ PCR amplification system
US648640122 Feb 200026 Nov 2002Tekcel, Inc.Multi well plate cover and assembly
US6896848 *19 Dec 200024 May 2005Tekcel, Inc.Microplate cover assembly
US706397913 Jun 200220 Jun 2006Grace Bio Labs., Inc.Interface between substrates having microarrays and microtiter plates
US766257225 Aug 200616 Feb 2010Platypus Technologies, Llc.Compositions and liquid crystals
US773181114 Apr 20068 Jun 2010Angros Lee HAnalytic substrate coating apparatus and method
US773190920 Feb 20048 Jun 2010Grace Bio-Labs, Inc.Reaction surface array diagnostic apparatus
US773659422 Jan 200315 Jun 2010Grace Bio-Labs, Inc.Reaction surface array diagnostic apparatus
US78424997 Aug 200730 Nov 2010Platypus Technologies, LlcSubstrates, devices, and methods for cellular assays
US800663814 Apr 200630 Aug 2011Angros Lee HAnalytic substrate coating apparatus and method
US803430628 Sep 200511 Oct 2011Grace Bio-Labs, Inc.Reaction surface array diagnostic apparatus including a flexible microtitre plate
US80482457 Jun 20101 Nov 2011Angros Lee HAnalytic substrate coating method
US80484727 Jun 20101 Nov 2011Angros Lee HAnalytic substrate coating method
US819299423 Nov 20095 Jun 2012Angros Lee HMethod of applying a biological specimen to an analytic plate
US826861410 Nov 200418 Sep 2012Platypus Technologies, LlcMethod for assaying cell movement
US828782214 May 201016 Oct 2012Grace Bio-Labs, Inc.Reaction surface array diagnostic apparatus
US845011631 May 201228 May 2013Lee H. AngrosMethod of applying a biological specimen to an analytic plate
US847010926 Oct 201125 Jun 2013Lee H. AngrosAnalytic substrate coating method
US847026426 Oct 201125 Jun 2013Lee H. AngrosAnalytic substrate coating method
US851297414 Sep 201220 Aug 2013Platypus Technologies, LlcMethod for assaying cell movement
DE2743799A1 *29 Sep 19776 Apr 1978Byrnes SenSchutzhandschuhe und garn zu deren herstellung
DE19923584A1 *21 May 19997 Dec 2000Memorec Medical Molecular ResIncubation container for samples on object carriers, useful for carrying out polymerase chain reactions, comprising chamber formed by plate, cover and seal, containing reservoir to minimize evaporation
DE19923584C2 *21 May 199924 Jan 2002Memorec Medical Molecular ResInkubationssystem
EP0014007A1 *10 Jan 19806 Aug 1980J. Hinrich Dr. PetersBiological container
EP0239450A1 *26 Feb 198730 Sep 1987Robert CassouMethod and device for in vitro fecundation and incubator for fecundation, and cellular culture used in this device and this method
EP0681024A2 *27 Apr 19958 Nov 1995ERIE SCIENTIFIC COMPANY (a Delaware Corporation)Improved biological culture slide and method of making same
WO1998015656A1 *3 Oct 199716 Apr 1998Univ New MexicoIn situ hybridization slide processes
WO2002092778A2 *17 May 200221 Nov 2002Univ Leland Stanford JuniorDevice and method for three-dimensional spatial localization and functional interconnection of different types of cells
WO2004046337A2 *19 Nov 20033 Jun 2004Tejal Ashwin DesaiMultilayered microcultures
WO2008021071A2 *7 Aug 200721 Feb 2008Platypus Technologies LlcSubstrates, devices, and methods for cellular assays
WO2008147783A2 *21 May 20084 Dec 2008Boston Scient Scimed IncApparatus and method of performing high-throughput cell-culture studies on biomaterials
WO2009103416A1 *4 Feb 200927 Aug 2009Eth ZurichMultiwell culture plate for three-dimensional cultures
WO2010062310A1 *22 Oct 20093 Jun 2010Millipore CorporationBiological culture assembly
Classifications
U.S. Classification435/305.3
International ClassificationC12M1/20, C12M3/00
Cooperative ClassificationC12M23/12, C12M23/04
European ClassificationC12M23/12, C12M23/04