Using CultureWell™ Chambered Coverglass Inserts
EMS Catalog #70461
Note: Package contents are sterile, handle carefully to prevent contamination.
1. Aseptically remove a CultureWell plate from the package. (Re-close package by folding bag end closed and securing it with a small piece of lab tape). Remove plate cover to expose MultiWell chambers (Figure 1).
To culture cells in MultiWell chambers
2. Pipette cells into individual wells (Figure 2) in a volume of 30-40ul (8-well chambers). 90-100ul (4-well chambers) 300-400ul (2-well chambers). Allow the cells to attach for 1-24 hours in a humidified CO incubator.
Wells are spaced 9mm for use with multichannel or automated pipettors.
3. After cells attach, flood the tray with 15ml of culture me dium (Figure 3).
Alternately, plates and inserts may be flooded with a suspension of cells in medium (15ml) and incubated to allow cells to attach to coverslips.
4. Cultures may be sub-divided or isolated by applying “press-to-seal” silicone well dividers (sold separately), (Figure 4).
After cultivation, cells may be washed, fixed and stained in the plate using standard protocols.
To remove a MultiWell Insert from a plate.
5. Grasp the insert at the central top notch with forceps and pull slowly upward and away from you (at an angle close to the plate) to release the silicone backing from the plate surface (Figure 5).
Coverslips may be stained, washed, etc. using conventional staining dishes by attaching a stainless steel handle to the insert frame as shown (Figure 6).
To remove a gasketed coverslip from a MultiWell insert.
Coverslips with gaskets attached may be individually separated from an insert by first removing the insert from the plate as previously described (Figure 6). If desired, the notched edge of the insert may be rested on the edge of the plate as illustrated (Figure 7). Using forceps grasp the silicone backing affixed to the coverslip at the notch in the insert and pull slowily toward you while gently pushing the insert frame downward. (Figure 7).
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 8).
To mount coverslips on glass microscope slides, first remove the silicone backing and gasket by holding the sides of the coverslip and gently peeling away the silicone backing (Figure 9) and then the silicone gasket (Figure 10).
CultureWells are for laboratory use only.