Using CultureWell™ Chambered Coverslips
EMS Catalog #70460
To place a MultiWell coverslip into a plate
- Place on workbench as shown (Figure 1). Aseptically peel back sterile pouch and top plastic liner to expose coverslips. (Figure 2).
- Using sterile forceps, lift the coverslip by grasping the silicone as shown (Figure 3). Remove the lid from a CultureWell plate or sterile 100mm diameter culture dish. Place the coverslip into the plate as shown in Figure 4 with the silicone gasketed wells facing upward.
To culture cells in MultiWell coverslips
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.
To remove a MultiWell Coverslip from a plate.
Grasp the silicone with forceps and pull slowly upward (Figure 6).
Coverslip handling and mounting
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.
To Clean and re-use MultiWell gaskets.
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 chambered coverslips are for laboratory use only