What he Confocal Microscope Can do for You

Ownby, C.L.(1)

(1)Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University

The scanning laser confocal microscope is an essential tool for today’s studies of the biology of cells.  Basically a light microscope system, the confocal microscope uses lasers to excite specific fluorescent dyes that have been incorporated into cellular molecules of interest to cell biologists and other scientists.  In addition to increased sensitivity when compared to a typical epifluorescent microscope, confocal microscopy provides better resolution and also the ability to optically section the sample allowing for 3D reconstruction.  Dual and triple labeling studies are also possible with the confocal microscope and digital images are routine. Immunolabelling experiments permit the localization of one, two or several proteins of interest in the cell either separately or simultaneously.  Proteins of interest can be co-localized with fluorescent probes specific for cellular organelles to determine their cellular location and distribution under various conditions.  Membrane trafficking in cells can be studied with green fluorescent protein (GFP) and other related proteins and membrane potentials and ion fluxes can be studied using specific fluorescent dyes.  Fluorescence in situ hybridization (FISH) allows for gene mapping and gene expression studies.  More recently many experiments on live cells can be performed with the confocal microscope and its close relative, the two-photon microscope. The purpose of this presentation is to explain how the confocal microscope works, to give many examples of the types of studies that can be done using it and to show how it can help you in your research projects.

CORRESPONDENCE TO:

Ownby, C.L., Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Email: carla@cvm.okstate.edu