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STED nanoscopy of KK114 ‐stained pathogenic bacteria
Author(s) -
Lucidi Massimiliano,
Hristu Radu,
Nichele Lorenzo,
Stanciu George A.,
Tranca Denis E.,
Holban Alina Maria,
Visca Paolo,
Stanciu Stefan G.,
Cincotti Gabriella
Publication year - 2020
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.202000097
Subject(s) - sted microscopy , bacteria , microscopy , bacillus subtilis , bacterial cell structure , fluorescence microscope , biophysics , microbiology and biotechnology , staining , live cell imaging , biology , super resolution microscopy , chemistry , fluorescence , cell , biochemistry , stimulated emission , optics , genetics , laser , physics
Super‐resolution microscopy techniques can provide answers to still pending questions on prokaryotic organisms but are yet to be used at their full potential for this purpose. To address this, we evaluate the ability of the rhodamine‐like KK114 dye to label various types of bacteria, to enable imaging of fine structural details with stimulated emission depletion microscopy (STED). We assessed fluorescent labeling with KK114 for eleven Gram‐positive and Gram‐negative bacterial species and observed that this contrast agent binds to their cell membranes. Significant differences in the labeling outputs were noticed across the tested bacterial species, but importantly, KK114‐staining allowed the observation of subtle nanometric cell details in some cases. For example, a helix pattern resembling a cytoskeleton arrangement was detected in Bacillus subtilis . Furthermore, we found that KK114 easily penetrates the membrane of bacterial microorganism that lost their viability, which can be useful to discriminate between living and dead cells.