A Simple Method for Scanning Electron Microscope Preparation of Cells Grown in Multiwell Culture Plates

Abstract

The closeness of wells in multiwell tissue culture plates makes it difficult to remove individual ones without disturbing the adjacent wells. Moreover, critical point drying frequently introduces artifact into the culture monolayer grown on plastic substrate. These problems make it difficult to process such cultures for scanning electron microscopy. However, for cell kinetic and correlative morphological studies on primary cultures derived from 7,12-dimethylbenz(α)anthracene(DMBA)-induced mammary tumors, we have found that Falcon 24-well tissue culture plates are excellent for maintenance of cells and are convenient to use. By plating the cells in alternating, diagonally disposed wells and while the cells are still in the buffer, individual wells can be cut from a multiwell plate without disturbing the cells in adjacent wells. The isolated wells can be used to carry out scanning electron microscope preparation. The height of the well is reduced with a scalpel prior to critical point drying; the remaining well is useful as a handle in mounting the dried sample to stubs for subsequent coating and viewing. Critical point drying and coating of monolayer samples on Falcon plastic are described. The results do not suggest that any artifacts are introduced in our preparations.