Rapid method of cheese processing for microflora and microstructural studies by electron microscopy

Document Type

Article

Publication Date

1-1-2004

Abstract

Previous methods for cheese sample processing for electron microscopy involves several days. Our objective was to identify a suitable yet rapid method. The rapid method involved an initial fixation of cheese in a solution of 2% glutaraldehyde and 1% OsO4 in 0.05M cacodylate buffer for 10 min. This was followed by a second fixation in 2% glutaraldehyde and 2% OsO 4 in 0.05M buffer for an additional 20 min. Sample fixation by the rapid method was attained in a total of 30 min compared to 17h in the control, i.e., the previous method, which required 15 h (overnight) fixation in 1 % glutaraldehyde and 2 h fixation in 1 % OsO4. After fixation, en block staining with aqueous uranyl acetate was conducted for 30 min. This was followed by ethanol dehydration and infiltration in resin. Additionally, the rapid method was conducted with LR White resin compared to Spurr's epoxy resin in the control. The culture bacteria, protein matrix and dispersed fat globules were clearly visible and were unaltered by the rapid method.

Publication Source (Journal or Book title)

Journal of Rapid Methods and Automation in Microbiology

First Page

199

Last Page

206

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