Live Single-Cell Imaging of Polyploid Cells

Hematopoietic cells are non-adherent cells which are not suitable for live single cell imaging as it is difficult to follow a single cell for certain time point due to their suspension nature and motility. To image a single cell for a specified time period we used specialized Microgrid arrays (MGA-050-02 & MGA-125-02) from microsurfaces Pty Ltd that can hold single cell in individual Microgrid arrays for imaging the same cell for specified time period. After sorting, live polyploid cells were set in the Microgrid array plate following manufacturer specified protocol. Cells were imaged with an in-house Nikon Eclipse Ti-E inverted microscope. The microscope is equipped with automated stage control, a perfect focus, and an Okolab cage incubator to maintain stable humidity, temperature and CO₂ levels. This set up can image live cells for 6-day periods without a reduction in the growth rate or viability of cells [23 (link)]. Images were captured every 20 minutes over a 48-hour period in order to visualize reductive cell division.

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Islam S., Paek A.L., Hammer M., Rangarajan S., Ruijtenbeek R., Cooke L., Weterings E, & Mahadevan D. (2018). Drug-induced aneuploidy and polyploidy is a mechanism of disease relapse in MYC/BCL2-addicted diffuse large B-cell lymphoma. Oncotarget, 9(89), 35875-35890.

Publication 2018

Eclipse Ti-E inverted microscope cage incubator

Cell division Cells Cells viability Hematopoietic Humidity Microscope Motility Polyploid cells Single individual

Corresponding Organization : University of Arizona

Other organizations : PamGene (Netherlands)

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Variable analysis

independent variables

Use of specialized Microgrid arrays (MGA-050-02 & MGA-125-02) from microsurfaces Pty Ltd to hold single cells for imaging

dependent variables

Visualization of reductive cell division over a 48-hour period

control variables

Stable humidity, temperature and CO2 levels maintained using an Okolab cage incubator
Nikon Eclipse Ti-E inverted microscope with automated stage control and perfect focus

controls

No positive or negative controls were explicitly mentioned in the provided information.

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