Cell Biological Applications of Fluorescence Microscopy

Link: http://www.itg.uiuc.edu/publications/techreports/99-006/

Stephen Rogers
Manager, Microscopy Suite, Light and Confocal Microscopy

The purpose of this technical bulletin is to provide an introduction to fluorescence microscopy to newcomers to the technique, and to collect a variety of protocols for specific applications of fluorescence in one place. In this bulletin, I have tried to assemble some of the techniques and lore that I have found particularly useful.

Contents
Introduction 2
Fluorescence Microscopy 3
Specimen Preparation 4
Fixation 5
Permeabilization 6
Blocking 7
Mounting Media 8
Photobleaching 9
Techniques for Staining Specific Cellular Structures 10
Microtubules 10
Actin 11
DNA 12
Mitochondria 13
Golgi Apparatus 14
Endoplasmic Reticulum 15
Sample Fixation Protocols 16
Glutaraldehyde Fixation 16
pH-Shift/Formaldehyde Fixation 17
Recipes 18
References 19

Imaging Technology Group
Beckman Institute for Advanced Science and Technology
University of Illinois at Urbana-Champaign
405 N. Mathews, Urbana, IL 61801
Technical Report 99-006
Date Issued: 21 April 1999