This Article Full Text Full Text (PDF) All Versions of this Article: jhc.7A7370.2008v1 56/7/697    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Google Scholar Articles by Reipert, S. Articles by Wiche, G. PubMed PubMed Citation Articles by Reipert, S. Articles by Wiche, G. Social Bookmarking                      What's this?

Rapid Microwave Fixation of Cell Monolayers Preserves Microtubule-associated Cell Structures

Siegfried Reipert, Harald Kotisch, Bhuma Wysoudil and Gerhard Wiche

Department of Molecular Cell Biology, Max F. Perutz Laboratories, University of Vienna, Vienna, Austria

Correspondence to: Siegfried Reipert, Department of Molecular Cell Biology, Max F. Perutz Laboratories, Dr. Bohr-Gasse 9, University of Vienna, A-1030 Vienna, Austria. E-mail: siegfried.reipert{at}univie.ac.at

Microwave (MW) fixation has been suggested as a method to rapidly immobilize cellular dynamics for fine structural studies in the electron microscope. To show its suitability for studies on cell monolayers, one has to apply MW fixation systematically in correlation with samples on the light microscopy level. Examples for MW fixation of cell monolayers, however, are still rare. MW-accelerated fixation for relatively long periods of time (1–2 min) has been reported without showing its suitability at the fine structural level. Here, we provide a rapid MW fixation protocol for cell monolayers on a subminute time scale. The impact of the MW-accelerated glutaraldehyde fixation on temperature-sensitive cytoskeletal components such as microtubules was evaluated. For testing the effectiveness of MW-assisted primary fixation, saponin treatment of the monolayers was included. Simultaneous MW-accelerated fixation and extraction by saponin was necessary to achieve a gradual improvement in visualization of cytoskeletal aspects in association with cell junctions, mitochondria, and centrioles. To establish a valuable routine program for fine structural studies of resin-embedded cell models on substrata, a protocol combining MW fixation with automatic processing in a tissue processor is provided. (J Histochem Cytochem 56:697–709, 2008)

Key Words: microwave fixation • cell monolayers • saponin extraction • electron microscopy • microtubules • cell junctions

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact

The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 2008