Culture slide assembly

Abstract: The present invention is a chamber slide assembly having a base member, a cooperable vessel comprising a chamber and a neck wherein said vessel is removably attached to the base and means for separating the vessel from the base. Preferably, the assembly may be used for carrying out biological reactions or growth therein, such as growing tissue or cell cultures.

Description: This invention relates to an assembly and a method for using the assembly in cell culture production. In particular, the assembly comprises a chamber, a slide and means for separating the chamber from the slide. It is frequently desirable in medical laboratory practice as well as in biological research to grow various bacteria, cells or tissues in particular media and then to examine the resulting growth. A particular use of this technique is in. virology laboratories where host cells are grown and then used to detect viral activity. Typical apparatus useful for carrying out biological reactions or growth, such as growing tissue cultures is described in U.S. Patent No. 3,726,764. The apparatus in U.S. Patent No. 3,726,764 is a chamber attached to a glass slide with a liquid-impermeable seal. A special tool is wedged into the seal to separate the chamber from the glass slide. A common problem which has confronted users of typical apparatus as described in U.S. Patent No. 3,726,764 is that a separate tool to remove the chamber from the slide is not convenient and in spite of the high level of skill and care in separating the chamber and the slide, the potential for not shattering the glass slide is not always assured, and therefore, contamination of the cells on the slide is also not assured. The precharacterizing part of claim 1 refers to a culture slide assembly disclosed in WO-A-91/13638. This culture slide assembly comprises a culture flask having a bottom closed by a laminate of an aperture film. This film is positioned upon a continuous carrier layer that is removably bonded to the side walls of the flask. The laminate carrying a cell culture can be removed by peeling from the flask. US-A-4,715,911 discloses a culture slide assembly comprising a vessel positioned on a carrier plate. The vessel is connected to the carrier plate by friction welding. This weld is so weak that it brakes when the vessel is manually forced away from the base plate. EP 0 681 024 A1 shall be considered as comprised in the state of the art according to article 54 (3) EPC. This document discloses a culture slide assembly comprising a vessel and a base plate. The vessel which is open at the bottom is bonded to the base plate by an adhesive bonding for removably attaching the vessel to the base plate which is a slide. The slide comprises a hydrophobic layer which has realized properties and facilitates detachment of the vessel from slide. It is an object of the present invention to provide an improved culture slide assembly that better protects the person carrying out the procedure and is comperatively simple and inexpensive to manufacture as compared to available devices. The culture slide assembly of the present invention is defined by claim 1. The method for growing tissue cultures is defined by claim 9. Preferably, the chamber comprises a forward section and a rearward section and sidewalls that extend from a top surface to a bottom surface, wherein the bottom surface is removably mated with the base member. Most preferably, the bottom surface of the receptacle is removably attached to the base member with a liquid-impermeable seal. Preferably the liquid-impermeable seal may be comprised of a silicon or acrylate based composition as well as organopolysiloxane elastomers. A means for separating the base member from the chamber is most preferably a movable lever. Desirably the lever is attached adjacent to the rearward section of the chamber. The lever may be used to provide appropriate force between the chamber and the base member to separate them. Most preferably, the base member is a microscope slide. The base member and receptacle are most preferable made of glass or plastic. To carry out biological reactions or growths in the apparatus, a liquid tissue culture medium is placed into the receptacle chamber that is in contact with the base member. The medium is incubated to allow the tissue culture to grow and to attach to the base member. The liquid medium is then removed from the chamber, and the receptacle is removed from the base with assistance from the lever. The tissue culture growth on the base can then be treated as desired and microscopically examined. This apparatus is also useful as an anaerobic chamber or as a blood culture chamber for microbiological assays. A feature of the invention is its ease of use, in that the receptacle is easily removable from the base by the lever without the need to have a separate tool to wedge between the chamber and the slide and that detachment forces are substantially reduced. Another feature of the invention is that the assembly is substantially self-contained in that the lever is attached to the assembly. Also, since the apparatus may be made from optically clear plastic, it may be easily disposable. Furthermore, the apparatus of the present invention improves the methods for carrying out biological reactions on slides by maintaining the integrity of the biological reaction due to the means for separating the chamber from the base member. FIG. 1 is a perspective view of a culture slide apparatus. FIG. 2 is a cross sectional side view of the culture slide apparatus of FIG. 1 taken along lines 2-2. FIG. 3 is side view of the culture slide of FIG. 1 illustrating the movement of the lever. FIG. 4 illustrates the removal of the receptacle from the base member by the lever. FIG. 5 illustrates the removal of the lever at the perforated edges. The preferred apparatus of the present invention is illustrated in FIG. 1 wherein apparatus 10 comprises a base member 12, a receptacle 14, a lever 16 which is movable and a lid 18 . Base member 12 has a planar upper surface 20 and opposing ends 22 and 24 which are normal to surface 20. Upper surface 20 of base member 12 is flat to form a mating surface for receptacle 14. Base member 12 is preferably a microscope slide. Such microscope slide is preferably fabricated from soda glass which has been cleaned to remove oils, greases, surfactants, abrasives or other materials inhibitory to biological or bacterial growth. Alternatively, the microscope slide may be fabricated from plastic. Such plastic materials may include polystyrene and the like. Receptacle 14 is located near end 24 and mates against surface 20 of base member 12 so as to leave a substantial portion 30 of surface 20 exposed near end face 22. This free surface area 30 can be etched so as to render it capable of receiving appropriate identification markings, such as an appropriate label. Receptacle 14 comprises opposing longitudinal sidewalls 34 and 36 and opposing transverse sidewalls 38 and 40 that define an open bottom 42 and an open top 44 . The sidewalls extend from a top surface 46 to a bottom surface 48 . At the bottom surface is a peripheral outwardly directed flange 50 surrounding bottom opening 42. As shown in FIG. 2 lever 16 comprises a rearward end 52 , and a forward or working end 54 . Rearward end 52 and forward end 54 of lever 16 meet at pivot position 56. Rearward end 52 is attached to flange 50 along sidewall 38 of receptacle 14 . The lever is attached to the receptacle in such a way that it can be moved in a substantially vertical up or down direction. As shown in FIG. 2, lid 18 is capable of fitting over and closing the open top of the receptacle and is easily removable. Receptacle 14 is preferably formed from transparent polymer materials. Such materials include, but are not limited to, polystyrene, polypropylene, celluloid, polymethacrylate and polymethylmethacrylate. In the fabrication of the apparatus of the present invention, bottom surface 48 is removably attached to surface 20 of base member 12 near end face 22 by suitable means such as by an adhesive. Such adhesives include silicons or acrylates. The principal criteria for a useful adhesive is that it provides desired removably adhesive characteristics between receptacle 14 and base 12, provides a liquid-impermeable seal, is substantially non-toxic to biological material subsequently employed in the apparatus, does not act as a source of growth for undesirable microorganisms and reduces the amount of detachment forces required to separate the receptacle from the base. As shown in FIG. 3, the lever may be activated wherein manual force is applied to the working end toward the upper surface of the base so that the pivot position contacts the upper surface of the base. This motion allows the working end of the level to exert force on the flange and bottom surface of the chamber in an upward direction so as to disengage the chamber from the base. The receptacle may be removed from the based by using the lever as shown in FIG. 4. Lever 16 may be made of plastic or other flexible material. Optionally, the lever may be removed from the receptacle at the perforated edges 60 between rearward end 52 and sidewall 40 of the receptacle as shown in FIG. 5. The apparatus of the present invention may be employed in the following manner to grow for example tissue cultures. The lid is removed from the receptacle. The desired liquid tissue culture medium containing a suspension of cells to be grown is then placed into the receptacle. The apparatus is then placed in a suitable incubator and is incubated under well-known conditions to carry out the tissue culture growth. If desired, suitable treatment may be carried out on the cells and medium during this growth to achieve cytopathology changes in the cells. At the conclusion of the growth period a mass of tissue cells is attached to base. The tissue culture medium can then be removed from the chamber by aspiration. The receptacle is then removed from the base by applying manual force to the lever as shown in FIG. 3 and removing the receptacle as shown in FIG. 4. The mass of tissue cells attached to the base is then rinsed and fixed on the base, and the affixed tissue culture is then treated with an appropriate stain to stain the culture. While the chamber described above has four-sides, it is understood that apparatus of the present invention can employ more or less sides as well as other geometric shapes, such as circular. Further, it is understood that the present invention can contain any desired number of chambers.

Claims: A culture slide apparatus comprising a base member (12), a receptacle chamber (14) and means for removably attaching said chamber (14) to said base member (12) characterized by a movable lever (16) attached to said chamber (14) for separating said chamber (14) from said base member (12). The apparatus of Claim 1, wherein said means for removably attaching said chamber (14) to said base member (12) is an adhesive material. The apparatus of Claim 2, wherein said adhesive provides a liquid-impermeable seal between said chamber (14) and said base member (12). The apparatus of Claim 1 wherein said base member (12) comprises a planar upper surface (20) and opposing ends (22,24) which are normal to said surface. The apparatus of Claim 1 wherein said chamber comprises top (46) and bottom surfaces (48), forward and rearward sections (38,40) and sidewalls (34,36) that extend from said top surface (46) to said bottom surface (48) wherein said bottom surface is removably attached with said base member (12). The apparatus of Claim 1 wherein said lever (16) comprises a rearward end (52) and a forward end (54) wherein said rearward and forward ends meet at a pivot position (56). The apparatus of Claim 6 wherein said lever (16) is removably attached to said chamber (14) by perforated edges (60). The apparatus of Claim 1 further comprising a removable lid (18). A method for growing tissue cultures with an apparatus having a base member (12), a receptacle chamber (14) removably attached to said base member, a movable lever (16) attached to said chamber and a removable lid (18), comprising: a) placing a liquid tissue culture medium containing a suspension of cells in said chamber (14); b) placing said lid (18) over said chamber (14), c) incubating said apparatus until a mass of tissue cells is attached to said base (12); d) removing said receptacle chamber (14) from said base (12) with said lever (16); e) rinsing said tissue cells attached to said base; f) staining said tissue cells; and g) examining said tissue cells.

Inventors: STEVENS TIMOTHY A, TYNDORF TADEUSZ A, STEVENS, TIMOTHY A., TYNDORF, TADEUSZ A.
Applicants: BECTON DICKINSON CO, BECTON, DICKINSON AND COMPANY
Application year: 1996
Main classification: C12M1
IPC codes: C12M1, G01N33, C12M3, G01N1