journal article·2005-12

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

Gary D. Josephsen, Kelly A. Josephsen, Greg J. Beilman, Jodie H. Taylor, Kristine E. Muiler

Microscopy and Microanalysis· Volume 11· Issue 6
Publisher page

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.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Hemodynamic Monitoring and Therapy · Non-Invasive Vital Sign Monitoring · Renal function and acid-base balance

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

References (15)

Show source-provided references
1. Trump, B.F. , Berezesky, I.K. , Laiho, K.U. , Osornio, A.R. , Mergner, W.J. , & Smith, M.W. (1980).The role of calcium in cell injury. A review.Scan Electron Microsc 492,437–462.
2. Fink, M. (2001).Cytopathic hypoxia, mitochondrial dysfunction as a mechanismcontributing to organ dysfunction in sepsis.Crit Care Clin 17,219–237.10.1016/S0749-0704(05)70161-5 ↗
3. Russin, W.A. & Trivett, C.L. (2001).Vacuum-assisted microwave processing of animal tissues for electronmicroscopy. InMicrowave Techniques and Protocols, Giberson, R.T. & Demaree, R.S. (Eds.), pp.25–35.Totowa, NJ:Humana Press.
4. Giberson, R.T. & Demaree, R.S. (1999).Microwave processing techniques for electron microscopy: A fourhour protocol. InMethods in Molecular Biology, Hajibagheri, N. (Ed.), pp.145–158.Totowa, NJ:Humana Press.10.1385/1-59259-201-5:145 ↗
5. Giberson, R.T. & Demaree, R.S., Jr. (1995).Microwave fixation: Understanding the variables to achieve rapidreproducible results.Microsc Res Tech 2,246–254.10.1002/jemt.1070320307 ↗
6. Sanders, M.A. & Gartner, D.M. (2001).In vivo microwave-assisted labeling of allium and drosophilanuclei. InMicrowave Techniques and Protocols. Giberson, R.T. & Demaree, R.S. (Eds.), pp.155–164.Totowa, NJ:Humana Press.
7. Giberson, R.T. , Demaree, R.S. Jr. , & Nordhausen, R.W. (1997).Four-hour processing of clinical/diagnostic specimens forelectron microscopy using microwave technique.J Vet Diagn Invest 9,61–67.10.1177/104063879700900111 ↗
8. Moore, F.A. , Haenel, J.B. , & Moore, E.E. (1995).Early impaired oxygen consumption due to supply-independentmitochondrial dysfunction occurs in patients who develop Multiple OrganFailure.Surg Forum 46,74–76.
9. Taylor, J.H. , Beilman, G.J. , Conroy, M.J , Mulier, K.E. , Myers, D. , Gruessner, A. , & Hammer, B.E. (2004).Tissue energetics as measured by nuclear magnetic resonancespectroscopy during hemorrhagic shock.Shock 21,58–64.10.1097/01.shk.0000101674.49265.93 ↗
10. Crouser, E.D. , Julian, M.W. , Blaho, D.V. , & Pfeiffer, D.R. (2002).Endotoxin-induced mitochondrial damage correlates with impairedrespiratory activity.Crit Care Med 30,276–284.10.1097/00003246-200202000-00002 ↗
11. Cairns, C.B. (2001).Rude unhinging of the machinery of life: Metabolic approaches tohemorrhagic shock.Curr Opin Crit Care 6,437–443.10.1097/00075198-200112000-00011 ↗
12. Giberson, R.T. (2001).Vacuum-assisted microwave processing of animal tissues for electronmicroscopy. InMicrowave Techniques and Protocols, Giberson, R.T. & Demaree, R.S. (Eds.), pp.155–164.Totowa, NJ:Humana Press.
13. Fox, C.H. , Johnson, F.B. , Whiting, J. , & Roller, P.P. (1985).Formaldehyde fixation.J Histochem Cytochem 8,845–853.10.1177/33.8.3894502 ↗
14. Kok, L.P. & Boon, M.E. (1997).Microwave methods for sample preparation in pathology. InMicrowave-Enhanced Chemistry: Fundamentals, Sample Preparation, andApplication, Kingston, H.M. & Haswell, S.J. (Eds.), pp.644–645.Washington, DC:American Chemical Society.
15. Cairns, C.B. , Moore, F.A. , Haenel, J.B. , Gallea, B.L. , Ortner, J.P. , Rose, S.J. , & Moore, E.E. (1997).Evidence for early supply independent mitochondrial dysfunction inpatients developing multiple organ failure after trauma.J Trauma 42,532–536.10.1097/00005373-199703000-00023 ↗