US3572892A - Multiple well tissue culture slide - Google Patents

Multiple well tissue culture slide Download PDF

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Publication number
US3572892A
US3572892A US797822A US3572892DA US3572892A US 3572892 A US3572892 A US 3572892A US 797822 A US797822 A US 797822A US 3572892D A US3572892D A US 3572892DA US 3572892 A US3572892 A US 3572892A
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US
United States
Prior art keywords
plate
wells
tissue culture
slide
depressions
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.)
Expired - Lifetime
Application number
US797822A
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English (en)
Inventor
Don P Metzgar
James V Sorrentino
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.)
Richardson Vicks Inc
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Richardson Merrell Inc
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Publication date
Application filed by Richardson Merrell Inc filed Critical Richardson Merrell Inc
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Publication of US3572892A publication Critical patent/US3572892A/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/04—Flat or tray type, drawers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/12—Well or multiwell 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
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides

Definitions

  • each well has an opening extending from the top to the bottom of the well adjacent to an edge of the plate to facilitate the drainage of liquids therefrom.
  • the bottom surface of each well is substantially parallel with the bottom surface of the plate.
  • This invention relates to a multiple well tissue culture slide. More particularly, this invention relates to a multiple well tissue culture microscope slide and to such a slide having fixed, confluent, viral infected tissue culture cells which can be used for the rapid diagnosis of viral diseases using fluorescent antibody technique.
  • Tissue culture cells of the required type and species are placed on a culture surface or vessel such as that of a Petri dish, microscope slide or microscope coverslip. After a sufficient period of incubation in which the cells become attached to the culture surface, the cells are infected with a specific virus agent. After another period of incubation, during which the virus agent multiplies, the culture vessel is removed from the incubator, the cells are fixed with a suitable fixative and then dried. This then becomes the basic unit for the fluorescent antibody technique as generally practiced in the prior art.
  • the fixed, infected tissue culture vessels are prepared in multiples. If serum dilutions are to be used a separate Petri dish, microscope slide or coverslip culture must be prepared for each dilution. If more than one serum sample is to be used, this requires multiples of the culture.
  • Each culture is then treated with a dilution of the patients or animals serum. The serum is allowed to remain in contact with the culture for 15 to 30 minutes. It is then rinsed in a suitable diluent to remove the excess serum. The culture is then treated for 15 to 30 minutes with a fluorescein conjugated homologous antiserum. Usually this antiserum is gamma globulin common to the species being tested. Each culture is rinsed to remove the excess fluorescein conjugate and then dried.
  • the culture After drying, the culture is examined microscopically, with a fluorescent microscope. A positive diagnosis is indicated by the presence of specific fluorescence in the treated culture.
  • tissue culture slide in which the wells contain a fixed, confluent sheet of cells infected with virus.
  • a slide can be supplied to a technologist or clinician performing antiviral tests, and thus further decrease the time and manipulative steps required for making tests on patients or animals.
  • the multiple well tissue culture slide of this invention comprises a transparent flat plate having depressions or wells on the top surface thereof.
  • the walls defining each of the wells have an opening extending from the top to the bottom of the well adjacent to an edge'of the plate in order to facilitate the drainage of liquids therefrom.
  • the bottom surface of the plate is substantially flat and preferably the bottom surface of each well is also flat and substantially parallel with the bottom surface of the plate.
  • this invention makes available, in one single slide, all of the basic materials required to perform the fluorescent antibody technique.
  • This invention can reduce the total time required for such diagnostic tests, including the preparative work, from 7 to 14 days to a maximum time of 2 hours.
  • an entire test requiring 6 or even more dilutions can be performed on a single slide. This eliminates the processing of multiple cultures and the necessity of individual microscopic examination of many cultures.
  • the multiple well slide can be examined in one microscopic manipulation.
  • Any virus agent that can be grown in tissue culture can be adapted to use with this invention.
  • the physician, laboratory or institution will have available a rapid and specific tool to be used in diagnosis of diseases of virus etiology.
  • FIG. 1 is a perspective view of the slide showing the top and sides thereof;
  • FIG. 2 is a perspective view of the slide of FIG. 1 showing the top and sides thereof with a confluent sheet of virus-in fected tissue culture cells in each of the wells;
  • FIG. 3 is a perspective view showing the bottom and sides of the slide of FIG. I.
  • the microscope slide 10 is a rectangular plate of a flat piece of glass or other transparent material such as a synthetic resin, e.g., polystyrene.
  • the slide 10 has two long sides or edges 12-12 parallel to each other; narrow sides or edges 1%14, at right angles to the long sides 12-12.
  • Each side 12 is millimeters (mm.) long.
  • Each narrow side 14 has a width of 25 mm.
  • a longitudinal rib 16 having a width of 5 mm. and transverse ribs 18 having a width of 2 mm. together with raised por tions along the narrow sides 14-14 define square wells or depressions 20 therebetween.
  • the top surfaces of ribs 16 and 18 and the raised portions along narrow sides 14-14 are in the same plane and parallel to slide bottom 26.
  • the thickness of the slide from the top surfaces of the ribs to the bottom surface 26 thereof is 1 mm.
  • Walls 24 enclose three sides of each well 20. These wells are 0.5 mm. deep and each well wall 24 has a length of 10 mm.
  • Wells 20 are open, from the bottom to the bottom thereof, adjacent their respective edge 12.
  • the bottom 26 of the microscope slide 10 is flat and parallel with the bottom 22 of each of the wells 20.
  • each of the wells 20 contains a fixed fluent virus-infected tissue culture 23 attached to the bottom surface 22 thereof.
  • the narrow ends 1444 form walls, 10 mm. wide, for one side of the wells at each end of the slide.
  • the slide has designators A and B for indicating each row of wells 22 and additionally has numerical designators 1-6 on ribs 16 between opposed wells so that the use of the letter and numerical designators can define each of the wells.
  • EXAMPLE 1 This example shows the use of the multiple well tissue culture slide of FIG. 1.
  • a single multiple well slide, as shown in FIG. 1, is prepared for the patients serum to be examined.
  • the slide is completely immersed in a culture vessel containing nutrient medium for the cells to be used.
  • human Wi-38 diploid cells were used.
  • Wi-38 cells are added to the medium covering the slide.
  • the multiple well culture is infected by adding an inoculum of adenovirus type 3 to the culture medium.
  • the virus infected multiple well slide is removed from the nutrient medium and fixed with acetone for 10 minutes and then dried.
  • the culture is attached to all the surfaces of the plate.
  • the culture is wiped off of the plate, except for that in the bottom of the wells 22 as appears as 28 in FIG. 2.
  • Each of the wells now contain a fixed confluent sheet of Wi-38 cells infected with adenovirus type 3.
  • Serum is withdrawn from the convalescing patient wherein adenovirus type 3 is suspected as a cause of the illness. Dilutions of the patients serum are prepared and each well is covered with about 0.1 milliliter of a single dilution.
  • One row of wells, e.g., Row A of FIG. 2, containing the control serum (normal serum) dilutions and one row, e.g., Row B of FIG. 2, of wells containing the patients serum, are prepared. Dilutions are prepared in two-fold serial dilutions of 1:2, 4, 8, l6 and 32.
  • the multiple well culture is then placed in a moist chamber for 15 to 30 minutes at 35 C. to 37 C. and then rinsed with 0.02 Molar phosphate buffered saline solution at pH 7.2 to remove the excess serum.
  • the multiple well culture is then examined microscopically in a fluorescent microscope, for the presence of specific fluorescence. lf positive, the preparation will fluoresce green, if negative, no fluorescence will be seen, as in the case of the normal serum controls.
  • the above test is carried out with a single slide and is observed microscopically in a single operation.
  • the normal serum and suspected serum are side by side, allowing for simultaneous observation.
  • EXAMPLE 2 This example shows a test to determine the presence of adenovirus antibody in a patients serum.
  • a kit containing 8 slides such as shown in FIG. 2 is supplied to the physician or laboratory.
  • Each slide has a confluent cell tissue culture infected with a different adenovirus type which has been fixed, as described in Example 1, paragraphs A and B. in this instance, however, the adenovirus is that of types I, 2, 5, 3, 4, 7, l4 and 2 l.
  • the physician treats each slide with the human suspected serum and with the normal serum as described in paragraph C of Example 1 and, by following the remaining procedure of Example 1, can determine within about 2 hours if one of the adenovirus types being screened was involved in the patients illness.
  • a multiple well tissue culture microscope slide comprisa. a flat plate of transparent material
  • said wells having an opening on one side thereof adjacent an edge of said plate, said opening extending from top to bottom of the well to facilitate drainage of liquids therefrom; and wherein d. the bottom surface of said plate opposite each well is flat.
  • a multiple well tissue culture microscope slide comprisa. a flat plate of transparent material
  • each of said depressions defined by sidewalls and a bottom;
  • said sidewalls having an opening adjacent one of the plate edges, said opening extending from the top to the bottom of said wells to facilitate drainage of liquids therefrom.
  • a multiple well tissue culture microscope slide comprisa. a flat plate of transparent material
  • said ribs enclosing a major portion of the wells and having an opening extending from the top to the bottom ofthe wells adjacent a plate edge to permit fluid communication between the well and said plate edge to facilitate drainage of liquids therefrom;
  • a multiple well tissue culture microscope slide comprisa. a transparent, flat rectangular plate
  • each row of wells having an opening extending from the bottom surface thereof to the top facing the adjacent plate edge, said openings facilitating the drainage of fluid out of said wells.
  • a multiple well tissue culture slide of claim 4 having attached to the bottom surface of said wells a fixed, confluent sheet of cells infected with virus.
US797822A 1969-02-10 1969-02-10 Multiple well tissue culture slide Expired - Lifetime US3572892A (en)

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US79782269A 1969-02-10 1969-02-10

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Country Status (8)

Country Link
US (1) US3572892A (de)
BE (1) BE745667A (de)
BR (1) BR6915183D0 (de)
DE (2) DE2005543A1 (de)
ES (1) ES376407A1 (de)
FR (1) FR2033318B1 (de)
GB (1) GB1248193A (de)
NL (1) NL7001767A (de)

Cited By (22)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3768914A (en) * 1972-09-18 1973-10-30 T Kinney Electron microscopy tissue grid staining and storing rack and method
US3862793A (en) * 1973-04-06 1975-01-28 Linbro Chemical Co Inc Device for quick alignment of any among grouped specimen areas on a plate with optical microscope axis
US4149803A (en) * 1975-05-23 1979-04-17 Litz Per Erik Composite petrographic thin section slide and method of making same
FR2533324A1 (fr) * 1982-09-20 1984-03-23 V Tech Inc Lame pour examen microscopique
FR2563010A1 (fr) * 1984-04-13 1985-10-18 Juglar Michel Lame pour immunofluorescence et son procede de fabrication
US4607921A (en) * 1982-09-20 1986-08-26 V-Tech, Inc. Wet mount microscopic examination slide II
FR2581194A1 (fr) * 1985-04-29 1986-10-31 Materiel Biomedical Dispositif pour l'analyse d'un liquide biologique.
FR2679659A1 (fr) * 1991-07-23 1993-01-29 Medgenix Diagnostic Sa Procede de dosage in vitro de molecules au niveau de leur site de production par des cellules en culture et format technologique adapte.
DE4417079A1 (de) * 1994-05-17 1995-12-07 Fraunhofer Ges Forschung Objektträger für Mikroskop
US5518925A (en) * 1995-06-06 1996-05-21 Becton Dickinson Co Culture slide assembly
US5605813A (en) * 1995-06-06 1997-02-25 Becton, Dickinson And Company Culture slide assembly
US5618731A (en) * 1995-06-06 1997-04-08 Becton, Dickinson And Company Culture slide assembly
USD378781S (en) * 1995-06-06 1997-04-08 Becton, Dickinson And Company Culture slide
USD382062S (en) * 1995-06-06 1997-08-05 Becton, Dickinson And Company Culture slide
US6121052A (en) * 1996-06-27 2000-09-19 Perkinelmer International C.V. Reflectance sampler and method of use
US6180314B1 (en) * 1998-05-27 2001-01-30 Becton, Dickinson And Company Method for preparing thin liquid samples for microscopic analysis
US6358475B1 (en) * 1998-05-27 2002-03-19 Becton, Dickinson And Company Device for preparing thin liquid for microscopic analysis
US20030109059A1 (en) * 2001-12-12 2003-06-12 Adrien Christopher L. Cover slip
US20030107946A1 (en) * 2001-10-25 2003-06-12 Cosby N. Guy Cover slip mixing apparatus and method
US20030197927A1 (en) * 2001-01-12 2003-10-23 Meus S.R.L. Slide for a microscopic examination of biological fluid
US20150370060A1 (en) * 2014-06-23 2015-12-24 Resolution Biomedical, Inc. Microscope slide with etched shapes
US11351551B2 (en) * 2017-11-03 2022-06-07 Illumina, Inc. Multi-well plate adaptors

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NO20004883D0 (no) * 2000-09-28 2000-09-28 Radiumhospitalets Forskningsst Objektglass, objektglass-sett, fremgangsmÕte for Õ frembringe prøvekjerner fra et objektglass, samt fremgangsmÕte for analyse av et prøveobjekt

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1516896A (en) * 1922-10-31 1924-11-25 Turner Edwin Branch Sampling device
US2281617A (en) * 1939-10-21 1942-05-05 Ramsdell Bentley Franklin Cigarette chest
US2302830A (en) * 1940-10-30 1942-11-24 Sol A Axelrad Microscope test slide
US3031924A (en) * 1959-03-12 1962-05-01 James C Lamal Observation slide
US3432275A (en) * 1964-08-31 1969-03-11 Hans Peter Olof Unger Display slide for wet biological preparates

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB692082A (en) * 1948-12-09 1953-05-27 Joachim Levitin Improvements in or relating to apparatus for measuring particle size, shape-factor, specific surface, and concentration of powdered or dispersed materials

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US1516896A (en) * 1922-10-31 1924-11-25 Turner Edwin Branch Sampling device
US2281617A (en) * 1939-10-21 1942-05-05 Ramsdell Bentley Franklin Cigarette chest
US2302830A (en) * 1940-10-30 1942-11-24 Sol A Axelrad Microscope test slide
US3031924A (en) * 1959-03-12 1962-05-01 James C Lamal Observation slide
US3432275A (en) * 1964-08-31 1969-03-11 Hans Peter Olof Unger Display slide for wet biological preparates

Cited By (27)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3768914A (en) * 1972-09-18 1973-10-30 T Kinney Electron microscopy tissue grid staining and storing rack and method
US3862793A (en) * 1973-04-06 1975-01-28 Linbro Chemical Co Inc Device for quick alignment of any among grouped specimen areas on a plate with optical microscope axis
US4149803A (en) * 1975-05-23 1979-04-17 Litz Per Erik Composite petrographic thin section slide and method of making same
FR2533324A1 (fr) * 1982-09-20 1984-03-23 V Tech Inc Lame pour examen microscopique
US4607921A (en) * 1982-09-20 1986-08-26 V-Tech, Inc. Wet mount microscopic examination slide II
FR2563010A1 (fr) * 1984-04-13 1985-10-18 Juglar Michel Lame pour immunofluorescence et son procede de fabrication
FR2581194A1 (fr) * 1985-04-29 1986-10-31 Materiel Biomedical Dispositif pour l'analyse d'un liquide biologique.
FR2679659A1 (fr) * 1991-07-23 1993-01-29 Medgenix Diagnostic Sa Procede de dosage in vitro de molecules au niveau de leur site de production par des cellules en culture et format technologique adapte.
WO1993002361A1 (fr) * 1991-07-23 1993-02-04 Medgenix Diagnostic S.A. Dosage in vitro de molecules au niveau de leur site de production par des cellules en culture
DE4417079A1 (de) * 1994-05-17 1995-12-07 Fraunhofer Ges Forschung Objektträger für Mikroskop
US5851489A (en) * 1994-05-17 1998-12-22 Micronas Intermetall Gmbh Specimen slide for a microscope
DE4417079C2 (de) * 1994-05-17 1998-06-10 Fraunhofer Ges Forschung Objektträger zum Beobachten von biologischem Material
US5618731A (en) * 1995-06-06 1997-04-08 Becton, Dickinson And Company Culture slide assembly
USD378781S (en) * 1995-06-06 1997-04-08 Becton, Dickinson And Company Culture slide
USD382062S (en) * 1995-06-06 1997-08-05 Becton, Dickinson And Company Culture slide
US5605813A (en) * 1995-06-06 1997-02-25 Becton, Dickinson And Company Culture slide assembly
US5518925A (en) * 1995-06-06 1996-05-21 Becton Dickinson Co Culture slide assembly
US6121052A (en) * 1996-06-27 2000-09-19 Perkinelmer International C.V. Reflectance sampler and method of use
US6358475B1 (en) * 1998-05-27 2002-03-19 Becton, Dickinson And Company Device for preparing thin liquid for microscopic analysis
US6180314B1 (en) * 1998-05-27 2001-01-30 Becton, Dickinson And Company Method for preparing thin liquid samples for microscopic analysis
US20030197927A1 (en) * 2001-01-12 2003-10-23 Meus S.R.L. Slide for a microscopic examination of biological fluid
US20030107946A1 (en) * 2001-10-25 2003-06-12 Cosby N. Guy Cover slip mixing apparatus and method
US6939032B2 (en) 2001-10-25 2005-09-06 Erie Scientific Company Cover slip mixing apparatus
US20030109059A1 (en) * 2001-12-12 2003-06-12 Adrien Christopher L. Cover slip
US7943093B2 (en) 2001-12-12 2011-05-17 Erie Scientific Company Cover slip
US20150370060A1 (en) * 2014-06-23 2015-12-24 Resolution Biomedical, Inc. Microscope slide with etched shapes
US11351551B2 (en) * 2017-11-03 2022-06-07 Illumina, Inc. Multi-well plate adaptors

Also Published As

Publication number Publication date
DE7004220U (de) 1971-09-02
DE2005543A1 (de) 1970-08-27
FR2033318A1 (de) 1970-12-04
BE745667A (fr) 1970-08-10
NL7001767A (de) 1970-08-12
GB1248193A (en) 1971-09-29
ES376407A1 (es) 1972-04-01
BR6915183D0 (pt) 1973-03-13
FR2033318B1 (de) 1974-05-03

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