Tobacco Leaf - Critical Point Drying Protocol for SEM

Application Note for Leica EM CPD300 - Life Science Research

November 18, 2016

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 7.32 h
0.1M Sodium Cacodylate Buffer, pH 7.32x10 min.
1% aqueous OsO41-2 h
Destilled water3x10 min.
Ethanol series: 50%, 70%, 95%, 100%3x10 min. 

Leica EM CPD300 auto Program:

Coating:
Platinum: 15-20 nm

Fig.1: Trichomes with Stomata from tobacco leaf
Fig.2: Stomata from tobacco leaf

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