Nematode E. dianae - Critical Point Drying Protocol for SEM

Leisch, Nikolaus

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November 16, 2016 Article

Nematode E. dianae - Critical Point Drying Protocol for SEM

Application Note for Leica EM CPD300 - Life Science Research

Introduction
Species: Eubostrichus dianae
Critical point drying of nematode Eubostrichus dianae to detect the ectosymbiotic bacteria layer with subsequent gold coating and SEM analysis.

Procedure
Sample Holder:
Samples were placed into the filter discs and porous pots holder.

Authors

Nikolaus Leisch , Mag.

University of Vienna, Department of Ecogenomics and Systems Biology, Austria

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EM Sample Preparation

Fixation and Dehydration

2.5% Glutaraldehyde in 0.1M Cacodylate Buffer	2 h
0.1M Cacodylate Buffer	3x10 min.
1% OsO4 in 0.1M Cacodylate Buffer	4-12 h
0.1M Cacodylate Buffer	3x10 min.
Ethanol series: 30%, 50%, 70%, 80%, 80%, 90%, 90%, 100%, 100%	10 min.
1:1 Mix Ethanol / Acetone	10 min.
100% Acetone	10 min.