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Microwave Processing for Sample Preparation to Evaluate Mitochondrial Ultrastructural Damage in Hemorrhagic Shock

Published online by Cambridge University Press:  15 November 2005

Gary D. Josephsen ,
Kelly A. Josephsen ,
Greg J. Beilman ,
Jodie H. Taylor  and
Kristine E. Muiler
Gary D. Josephsen
Affiliation:
Section of Surgical Critical Care, Department of Surgery, University of Minnesota, Minneapolis, MN 55455, USA
Kelly A. Josephsen
Affiliation:
The Imaging Center, University of Minnesota, St. Paul, MN 55108, USA
Greg J. Beilman
Affiliation:
Section of Surgical Critical Care, Department of Surgery, University of Minnesota, Minneapolis, MN 55455, USA North Trauma Institute, North Memorial Health Care, Robbinsdale, MN 55422, USA
Jodie H. Taylor
Affiliation:
Section of Surgical Critical Care, Department of Surgery, University of Minnesota, Minneapolis, MN 55455, USA North Trauma Institute, North Memorial Health Care, Robbinsdale, MN 55422, USA Hennepin County Medical Center, Minneapolis, MN 55415, USA
Kristine E. Muiler
Affiliation:
Section of Surgical Critical Care, Department of Surgery, University of Minnesota, Minneapolis, MN 55455, USA
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Abstract

This is a report of the adaptation of microwave processing in the preparation of liver biopsies for transmission electron microscopy (TEM) to examine ultrastructural damage of mitochondria in the setting of metabolic stress. Hemorrhagic shock was induced in pigs via 35% total blood volume bleed and a 90-min period of shock followed by resuscitation. Hepatic biopsies were collected before shock and after resuscitation. Following collection, biopsies were processed for TEM by a rapid method involving microwave irradiation (Giberson, 2001). Samples pre- and postshock of each of two animals were viewed and scored using the mitochondrial ultrastructure scoring system (Crouser et al., 2002), a system used to quantify the severity of ultrastructural damage during shock. Results showed evidence of increased ultrastructural damage in the postshock samples, which scored 4.00 and 3.42, versus their preshock controls, which scored 1.18 and 1.27. The results of this analysis were similar to those obtained in another model of shock (Crouser et al., 2002). However, the amount of time used to process the samples was significantly shortened with methods involving microwave irradiation.

Keywords

microwavemitochondriaultrastructurehemorrhagic shocktransmission electron microscopy
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 11 , Issue 6 , December 2005 , pp. 500 - 505
Copyright
© 2005 Microscopy Society of America

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