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Immunofluorescence assays were performed and analysed by confocal microscopy using the antibodies described below.
Technical replicates consisted of aliquots of E. coli cultures that were independently prepared for microscopy using the LIVE/DEAD protocol.
Ca2+ levels in the cytosol of FDB fibers was monitored via fluorescence microscopy using the calcium-sensitive probe Fura-2 essentially as described previously77.
Martell, J. D., Deerinck, T. J., Lam, S. S., Ellisman, M. H. & Ting, A. Y. Electron microscopy using the genetically encoded APEX2 tag in cultured mammalian cells.
Cell viability was assessed by live-cell confocal microscopy using the Nuclear-ID™ Blue/Red viability assay (Enzo Life Sciences) according to the manufacturer's recommendations.
RM, SM and Golgi fractions were prepared for electron microscopy using the random sampling filtration protocol detailed previously (Bell et al., 2001).
Following 0 and 7 h, cells were prepared for fluorescent microscopy using the LIVE/DEAD Baclight Bacteria Viability Kit (Thermo Fisher) to characterize the fraction of viable cells across the population.
The dislocation arrangement and the dissociation of superdislocations were studied by transmission electron microscopy using the weak-beam technique.
Morphology of samples was studied by transmission electron microscopy using the Tecnai G2 20X-TWIN USA microscope.
Immunofluorescent images were confirmed with immunoelectron microscopy using the same monoclonal antibody (Figure 8 A B).
Images were acquired by automated fluorescence microscopy using the Cellomics ArrayScan VTI (Thermo Fisher).
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