Protocol detail

TEM Analysis of Infected Macrophages

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For TEM analysis, infected macrophages were fixed with a solution containing 2% (wt/vol) paraformaldehyde (Merck, Ireland) and 2.5% (vol/vol) glutaraldehyde (Merck, Ireland) in 0.1 M phosphate buffer (Sigma–Aldrich, USA) at pH 7.4 for 1 h. After fixation, the infected macrophages were recovered using a cell scraper and were processed following conventional procedures (Lee et al., 2011 (link)). The sections were observed using a Jeol 1400 transmission electron microscope (Jeol, Japan).

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Brambilla C., Llorens-Fons M., Julián E., Noguera-Ortega E., Tomàs-Martínez C., Pérez-Trujillo M., Byrd T.F., Alcaide F, & Luquin M. (2016). Mycobacteria Clumping Increase Their Capacity to Damage Macrophages. Frontiers in Microbiology, 7, 1562.

Publication 2016

paraformaldehyde glutaraldehyde 0.1 M phosphate buffer

Buffer Cell Glutaraldehyde Macrophages Paraformaldehyde Phosphate Transmission electron microscope

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Variable analysis

independent variables

Fixation solution containing 2% (wt/vol) paraformaldehyde and 2.5% (vol/vol) glutaraldehyde in 0.1 M phosphate buffer at pH 7.4 for 1 h

dependent variables

Infected macrophages observed using a Jeol 1400 transmission electron microscope

control variables

0.1 M phosphate buffer (Sigma–Aldrich, USA) at pH 7.4

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