Cell Imaging and Analysis Protocol

For imaging, the cells were immobilized on 1% agarose [30 (link)] and photographed with a CoolSnap fx (Photometrics) charge-coupled device (CCD) camera mounted on an Olympus BX-60 fluorescence microscope through an UPLANFl 100×/1.3 oil objective (Olympus, Tokyo, Japan). Images were taken using modified acquisition software that used the program ImageJ by Wayne Rasband and analyzed using Object-J’s Coli-Inspector [17 (link)]. Statistical testing of normalized DAPI localization patterns was done using Kolmogorov–Smirnov testing available on http://www.physics.csbsju.edu/stats/ [31 ].

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Meiresonne N.Y, & den Blaauwen T. (2020). The In Vitro Non-Tetramerizing ZapAI83E Mutant Is Unable to Recruit ZapB to the Division Plane In Vivo in Escherichia coli. International Journal of Molecular Sciences, 21(9), 3130.

Publication 2020

UPLANFl 100×/1.3 oil objective

Agarose Cells Dapi Device Fluorescence microscope

Corresponding Organization : University of Amsterdam

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Variable analysis

independent variables

Immobilization of cells on 1% agarose

dependent variables

DAPI localization patterns

control variables

Olympus BX-60 fluorescence microscope
UPLANFl 100×/1.3 oil objective (Olympus, Tokyo, Japan)
CoolSnap fx (Photometrics) charge-coupled device (CCD) camera
Modified acquisition software using ImageJ by Wayne Rasband
Object-J's Coli-Inspector for image analysis

positive controls

No positive controls explicitly mentioned

negative controls

No negative controls explicitly mentioned

Annotations

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