TEM Sample Preparation for BMVs

Sample preparation for TEM was performed as described previously (11 (link)). Briefly, BMVs were coated onto carbon-coated 400 mesh copper grids (ProSciTech), fixed in 1% (wt/vol) glutaraldehyde (Sigma) in PBS, stained with 2% (wt/vol) uranyl acetate (ProSciTech) (pH 7.0), and coated with 2% (wt/vol) methyl-cellulose (Sigma) in 0.4% (wt/vol) uranyl acetate (pH 4.0). Samples were viewed using a JEOL JEM-2100 transmission electron microscope (JEOL, Japan) operated at 200 kV fitted with a Valeta 4 MP CCD camera (Emsis, Germany).

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Bitto N.J., Zavan L., Johnston E.L., Stinear T.P., Hill A.F, & Kaparakis-Liaskos M. (2021). Considerations for the Analysis of Bacterial Membrane Vesicles: Methods of Vesicle Production and Quantification Can Influence Biological and Experimental Outcomes. Microbiology Spectrum, 9(3), e01273-21.

Publication 2021

carbon-coated 400 mesh copper grids glutaraldehyde uranyl acetate methyl-cellulose JEM-2100 transmission electron microscope Valeta 4 MP CCD camera

Carbon Copper Glutaraldehyde Methyl cellulose Transmission electron microscope Uranyl acetate

Corresponding Organization : La Trobe University

Other organizations : University of Melbourne, Peter Doherty Institute

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Variable analysis

independent variables

Sample preparation method

dependent variables

Visualization of BMVs using TEM

control variables

Carbon-coated 400 mesh copper grids
Glutaraldehyde fixation
Uranyl acetate staining
Methyl-cellulose coating

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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