Tobacco Leaf - Critical Point Drying Protocol for SEM

November 18, 2016 Article

Application Note for Leica EM CPD300 - Life Science Research

Species:
Tobacco (Nicotiana tabacum)
Critical point drying of tobacco leafs with subsequent platinum coating and SEM analysis.

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

Fixation and Dehydration:

2% Paraformaldehyde, 2.5% Glutaraldehyde, 0.1M Cacodylate Buffer, pH 7.3	2 h
0.1M Sodium Cacodylate Buffer, pH 7.3	2x10 min.
1% aqueous OsO4	1-2 h
Destilled water	3x10 min.
Ethanol series: 50%, 70%, 95%, 100%	3x10 min.

Leica EM CPD300 auto Program:

Coating:
Platinum: 15-20 nm