Concepts And Tips For Better Fixed Sample Imaging Results

By Paul Held, Laboratory Manager, Applications Dept., BioTek Instruments, Inc.

Multiple processing steps are required to prepare tissue culture cells for fluorescence microscopy. Experiments are generally classified as being either live or fixed cell microscopy. Fluorescence microscopy of live cells uses either genetically encoded fluorescent proteins (e.g. GFP, mcherry, YFP, RFP, etc.) or cell membrane-permeable, non-toxic fluorescent stains. Fluorescence microscopy of fixed cells uses a fixative agent that renders the cells dead, but maintains cellular structure, allowing the use of specific antibodies and dyes to investigate cell morphology and structure. Appropriate sample preparation is necessary to ensure high quality images are captured. Here we describe a number of concepts and considerations regarding the sample preparation process that can assist with automated digital fluorescence microscopy of fixed cells.