Detection of cells grown on non-transparent supports: Determination of cell growth characteristics on fully transparent supports can easily be obtained by conventional phase contrast microscopy. However, with semi- and non-transparent supports the conventional methods to determine the appropriate parameters fail. For that reason we suggest a quick and practical approach for detection of cells on non-transparent supports with an epifluorescence microscope. See ref. [15].

Procedure takes 60 minutes or less:

1. For example, cells are cultured on the upper side of a support mounted on a tissue carrier so that the white tension ring can be seen.

2. Place the tissue carrier in a 24 well plate.

3. Fix the specimens in 70% ethanol for 10 minutes.

4. Stain in fluorescent DAPI or propidium iodide (4 mg/ml) in phosphate buffered saline without Ca²⁺ and Mg²⁺ for 5 respectively 30 minutes.

5. Wash 3 x 10 minutes in phosphate buffered saline.

6. Turn the tissue carrier upside down with fine forceps so that the cells are now on the lower side of the support within a 24 well culture plate. The white tension ring now faces the bottom of the plate.

7. Record the nuclear staining profile of the cells within phosphate buffered saline under the inverse epifluorescence microscope in a 24 well culture plate.