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Visualizing bleb mass dynamics in single cells using quantitative phase microscopy

Abstract

Understanding biological responses to directed energy (DE) is critical to ensure the safety of personnel within the Department of Defense. At the Air Force Research Laboratory, we have developed or adapted advanced optical imaging systems that quantify biophysical responses to DE. One notable cellular response to DE exposure is the formation of blebs, or semi-spherical protrusions of the plasma membrane in living cells. In this work, we demonstrate the capacity of quantitative phase imaging (QPI) to both visualize and quantify the formation of membrane blebs following DE exposure. QPI is an interferometric imaging tool that uses optical path length as a label-free contrast mechanism and is sensitive to the non-aqueous mass density, or dry mass, of living cells. Blebs from both CHO-K1 and U937 cells were generated after exposure to a series of 600 ns, 21.2 kV/cm electric pulses. These blebs were visualized in real time, and their dry mass relative to the rest of the cell body was quantified as a function of time. It is our hope that this system will lead to an improved understanding of both DE-induced and apoptotic blebbing.

© 2021 Optical Society of America

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Supplementary Material (4)

NameDescription
Visualization 1       CHO-K1 cell blebbing in response to pulsed electric fields. Scale bar 5 microns.
Visualization 2       CHO-K1 cell blebbing in response to pulsed electric fields. Scale bar 5 microns.
Visualization 3       U937 cell developing a massive bleb in response to pulsed electric fields. Scale bar 10 microns.
Visualization 4       U937 cell with multiple blebs merging into a single massive bleb. Scale bar 10 microns.

Data Availability

Data underlying the results presented in this paper are not publicly available at this time but may be obtained from the authors upon reasonable request.

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