Examination of whole cell mounts by transmission electron microscopy using cultures grown on carbon coated coverslips

Abstract

SUMMARY Cultures of porcine granulosa cells have been grown on serum coated carbon films deposited on glass coverslips. Intact and detergent‐treated cultures were critical point dried, chromium shadowed and removed from the coverslips by dilute hydrofluoric acid. Whole cell mounts were examined by conventional transmission electron microscopy. The advantage of this system over other methods includes the ability to: (1) examine cells with a highly rounded morphology, (2) use high cell densities, (3) study cell projections extending onto the substratum, (4) maintain cultures for long periods, (5) examine large samples easily, and (6) readily use the same cultures for transmitted light and fluorescent microscopy, scanning and transmission electron microscopy (SEM and TEM).

A simple technique for the visualization of whole mount cytoskeletons with transmission electron microscopy

“…Whether this technique is suitable for contracting cell models has yet to be demonstrated. Tolson and Boothroyd (1982) have presented a method similar to ours, using carbon-coated coverslips. Their technique requires glow discharging to remove surface charge from the carbon layer.…”