Black Mould Protocol

Application Note for Leica EM CPD300 - Life Science Research

August 09, 2016

Introduction
Species: Black mould (Aspergilus niger)
Critical point drying of Black mould with subsequent platinum / palladium coating and SEM analysis to detect conidiospores

Procedure
Sample Holder: Sample was inserted into a filter disc (Pore size: 16 - 40 μm) and placed into the filter discs and porous pots holder

Cultivation
Cultivate fungi on agar containing growth medium for 3 days

Fixation and Dehydration

3% Glutaraldehyde in PBS, pH 7.3 at 4°C18 h
Distilled water3x10 min.
1% aqueous OsO4, 4°C18 h
Distilled water3x10 min.
Ethanol series: 30%, 50%, 70%, 80%, 90%, 96%, 100% at 25°C1x10 min.
1% DMP in Acetone series: 30%, 50%, 100%3x30 min.

CPD300 auto program


Coating
Mount the dried samples on stubs containing carbon adhesives
Platinum / Palladium coating: 6 nm

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Fig.1: Black mould conidiospores
Fig.2: Black mould conidiospores

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