Paper Info
Title
Preparing Whole Mounts Of Biological Specimens For Imaging Macromolecular Structures By Light And Electron Microscopy
Abstract
In this article we critically review the procedures involved in preparing whole mounts of biological samples for microscopic observation, particularly at high resolution. We discuss practical methods for optimizing specimen preservation to achieve the two principal goals of biological specimen preparation: (a) preserving biological structures as close to their living configuration as possible, and (b) rendering them visible with the desired imaging method. Drawing on examples taken from our own work, and using fluorescence light microscopy and scanning electron microscopy to study the cytoskeleton of vertebrate cells in culture, we show that obtaining optimal results is often a compromise between maximum preservation and the clear visualization of structures. We have found that fixing cells with bifunctional protein crosslinking reagents, in combination with detergent extraction, allows internal cell structures to be preserved, labeled with specific probes, and visualized with microscopic methods. We also review and discuss the relative merits of different procedures for fixing (chemical fixation and cryofixation), drying (air-drying, critical point-drying, and freeze-drying) and coating biological specimens with metals to facilitate visualization in the electron microscope. (C) 1997 John Wiley & Sons, Inc.
Year
DOI
Venue
1997
10.1002/(SICI)1098-1098(1997)8:3<225::AID-IMA1>3.0.CO;2-C
INTERNATIONAL JOURNAL OF IMAGING SYSTEMS AND TECHNOLOGY
Keywords
Field
DocType
electron microscopy
Nanotechnology,Specimen preservation,Biological specimen,Computer science,Macromolecule,Scanning electron microscope,Cryofixation,Electron microscope,Fluorescence Light Microscopy
Journal
Volume
Issue
ISSN
8
3
0899-9457
Citations 
PageRank 
References 
0
0.34
0
Authors
2
Name
Order
Citations
PageRank
Paul B. Bell100.34
Barbara Safiejko-Mroczka200.34