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Pipette cells into individual wells (Figure 5) in a volume of 30-40ul (8-well chambers). 90-100ul (4-well chambers) or 300-400ul (2-well chambers). Allow the cells to attach for 1-24 hours in a humidified CO 2incubator. Wells are spaced 9mm for use with multichannel or automated pipettors.
After cells attach, flood the tray with 20ml of culture medium (Figure 5).
Alternately, plates and coverslips may be flooded with a suspension of cells in medium (20ml-Plates) and incubated to allow cells to attach to coverslips. Cells do not attach to the silicone gaskets.
Unused coverslips from opened pouches may be stored in sterile culture dishes to prevent contamination.
After cultivation, cells can be washed, fixed and stained in the plate or in staining dishes using standard protocols.
Grasp the silicone with forceps and pull slowly upward (Figure 6).
Coverslips are removed with the silicone gasket still attached to provide stability and protection from breakage. Individual gasketed coverslips may be placed in coplin jars, or any holder designed for standard microscope slides (Figure 7).
Multiwell coverslips may be placed in culture dishes for antibody / hybriziation, and washing steps, (Figure 8).
To mount coverslips on glass microscope slides, first remove the silicone gasket by peeling it off as shown (Figures 9).
Specimens may also be mounted using aqueous media by filling wells and pressing another coverslip to the gasket.
Rinse off reagents immediately after each use. Wash silicone gaskets using distilled water and if necessary mild detergent suitable for cell culture. Do not soak gaskets in detergent. Rinse thoroughly with distilled water and allow gaskets to dry. Just prior to applying a new coverslip, use a piece of
adhesive tape to clean lint and debris from the gasket surface.
Place a clean, dry 24mm x 50mm glass coverslip onto the silicone gasket and press gently to seal. Place gasketed coverslips individually, into glass petri dishes (silicone side up), wrap with aluminum foil and sterilize by autoclaving. Allow coverslips to cool and dry completely before use.
CultureWell™ MultiWell Chambered Coverslips are available online from the EMS Catalog. For ordering or product information, click here.