U.S. patent number 3,883,398 [Application Number 05/357,991] was granted by the patent office on 1975-05-13 for microculture slide chamber.
This patent grant is currently assigned to Bellco Glass, Inc.. Invention is credited to Kazuyuki Ray Ono.
United States Patent |
3,883,398 |
Ono |
May 13, 1975 |
Microculture slide chamber
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.
Inventors: |
Ono; Kazuyuki Ray (Bridgeton,
NJ) |
Assignee: |
Bellco Glass, Inc. (Vineland,
NJ)
|
Family
ID: |
23407855 |
Appl.
No.: |
05/357,991 |
Filed: |
May 7, 1973 |
Current U.S.
Class: |
435/305.3 |
Current CPC
Class: |
C12M
23/12 (20130101); C12M 23/04 (20130101); C12M
41/36 (20130101) |
Current International
Class: |
C12M
3/00 (20060101); C12M 1/16 (20060101); C12M
1/20 (20060101); C12b 001/00 () |
Field of
Search: |
;195/139,127 |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Tanenholtz; Alvin E.
Attorney, Agent or Firm: Seidel, Gonda & Goldhammer
Claims
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 second 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 relation 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.
Description
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 mono-layer 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:
1. 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. Immunoflourescence 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 patient's 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 maintaining 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 10 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 0.2 ml. The slide 12 is preferably of standard
dimensions for microscope slides which are approximately 0.9 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.sub.2 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 mono-layer.
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.
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