Water Flea Protocol

Application Note for Leica EM CPD300 - Life Science Research

June 07, 2016

Introduction:
Critical point drying of water flea with subsequent gold coating and SEM-Analysis to detect finde surface structures.

Procedure:
Sample Holder:
Sample was inserted into a filter disc
(Pore size: 16 - 40 μm).
Filter disc was places into the cover slip holder 18 mm.

Fixation and Dehydration:

2.5% Glutaraldehyde in 0.1M Sodium Cacodylate Buffer, 2% Sucrose, pH 7.31x 18 h
0.1 M Sodium Cacodylate Buffer, 2 % Sucrose, pH 7.33x 10 min.
0.1% OsO4, in 0.1M Sodium Cacodylate Buffer, 2% Sucrose, pH 7.31x 1 h
0.1 M Sodium Cacodylate Buffer, 2 % Sucrose, pH 7.33x 10 min.
Ethanol 30%, 50%, 70%, 80%, 90%, 96%, 100%2x 10 min.
100% Acetone, 1% Dimethoxypropane2x 30 min.

 

Leica EM CPD300 auto Program:

 

Coating:
Gold: 10-20 nm

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Water flea

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