2023
DOI: 10.1101/2023.05.15.540883
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Quantitative microbiology with widefield microscopy: navigating optical artefacts for accurate interpretations

Abstract: Live cell imaging of microbial cells with microscopy has revolutionised quantitative microbiology. Micrographs are one of the most information-rich data types captured about a microbe, allowing quantification of the size and morphology of individual cells and their gene-expression over time. However, an optical microscope is a diffraction-limited system, and the comparable size of the point spread function of the microscope to the size of a microbial cell can lead to imaging artefacts which corrupt and bias th… Show more

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Cited by 2 publications
(2 citation statements)
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“…This arrangement was not seen in the 2D projection ( Fig. 1A i ), highlighting that projection effects in widefield and 2D SMLM images might not reveal the actual organization of the target structure 7,39 . In the 3D image, the nucleoid appears positioned close to the cell membrane, as we have previously shown using STED microscopy 40 .…”
Section: Resultsmentioning
confidence: 90%
See 1 more Smart Citation
“…This arrangement was not seen in the 2D projection ( Fig. 1A i ), highlighting that projection effects in widefield and 2D SMLM images might not reveal the actual organization of the target structure 7,39 . In the 3D image, the nucleoid appears positioned close to the cell membrane, as we have previously shown using STED microscopy 40 .…”
Section: Resultsmentioning
confidence: 90%
“…RNL describes the fraction of cell length that is populated by chromosomal DNA. This metric is similar to the relative nucleoid size used by Cabrera and colleagues 47 , yet we prefer RNL in diffraction-limited images due to the 2D projection effects shown in Figure 1A and literature 7,39 . To monitor reorganization of MreB following antibiotic treatment, we quantified its relative distribution at the cell cylinder and cell poles (Fig.…”
Section: Inhibition Of Biosynthetic Processes Alters Nucleoid Organiz...mentioning
confidence: 89%