Image-based Single-Cell Analysis Protocol

Image acquisition was preformed using Cellometer® Vision CBA Image Cytometer (Nexcelom Bioscience LLC.). For image analysis we used Nexcelom Data Package and MATLAB software. Nexcelom filter VB-595-502 was used for red fluorescence assay in Cellometer. We used Nexcelom image segmentation (Supplementary Figs. 3 and 8) for object detection upon setting Cell Diameter parameter to 2–5 μm for single cells and 5–20 μm for clumps^{95 (link)}. The Object Roundness parameter was set to 0.45-0.8 (with 1.0 used for perfectly circular shapes) to separate single living cells from debris.

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Guinn L., Lo E, & Balázsi G. (2022). Drug-dependent growth curve reshaping reveals mechanisms of antifungal resistance in Saccharomyces cerevisiae. Communications Biology, 5, 292.

Publication 2022

Cellometer® Vision CBA Image Cytometer

Assay Cells Figs Fluorescence Vision

Corresponding Organization :

Other organizations : Stony Brook University

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Variable analysis

independent variables

Cell Diameter parameter (2–5 μm for single cells, 5–20 μm for clumps)
Object Roundness parameter (0.45-0.8)

dependent variables

Object detection

control variables

Nexcelom filter VB-595-502 for red fluorescence assay

controls

No positive or negative controls were explicitly mentioned.

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