Confocal scanning-laser microscopy combined with sophisticated image-processing techniques allow the cell biologist to explore the three-dimensional infrastructure of the cell, thus conferring a high degree of precision in localization experiments. This chapter discusses some of the inherent problems in preservation of biological specimens so that reliable data can be acquired with a confocal fluorescence microscope. The methodology is the result of an effort to study the genesis of epithelial polarity in the Madin-Darby canine kidney (MDCK) cell line. While specific methods have been developed to study these particular cells, the lessons learned about their preservation should be widely applicable. A description of the confocal principle introduces basic concepts of confocal fluorescence microscopy. One can then understand how images are recorded and how the information may be interpreted.
ASJC Scopus subject areas
- Cell Biology