Cell Volume Measurements by Optical Transmission Microscopy. Issue 1 (1st July 2019)

• Record Type: Journal Article
• Authors: Model, Michael A.
• Editors: Robinson, J. Paul; Darzynkiewicz, Zbigniew; Dobrucki, Jurek; Hyun, William C.; Nolan, John P.; Orfao, Alberto; Rabinovitch, Peter S.
• Is Part Of: Current protocols in cytometry. Volume 90:Issue 1(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2019-07-01
• Subjects: Acid Blue 9 -- cell volume -- optical microscopy -- transmission‐through‐dye microscopy; Cytology -- Laboratory manuals; Flow cytometry -- Laboratory manuals; Cell separation -- Laboratory manuals; Molecular biology -- Laboratory manuals; Flow Cytometry -- methods; Image Cytometry -- methods; Cell Separation -- methods; Cytological Techniques; Molecular Biology -- methods; Cell separation; Cytology; Flow cytometry; Molecular biology; Laboratory Manuals; Laboratory manuals; 571.6
• DOI: 10.1002/cpcy.62
• Languages: English
• ISSNs: 1934-9297

Abstract

Abstract: Cell volume is an important parameter in studying cell adaptation to anisosmotic stress, activation of monovalent ion channels, and cell death. This article describes a method for measurement of the volumes of adherent cells using a standard light microscope. A coverslip with attached cells is placed in a shallow chamber in a medium containing a strongly absorbing and cell‐impermeant dye, Acid Blue 9. When such a sample is imaged in transmitted light at a wavelength of maximum dye absorption (630 nm), the resulting contrast quantitatively reflects cell thickness; once the thickness is known at every point, the volume can be computed as well. Technical details, interpretation of data, and possible artifacts are discussed. © 2019 by John Wiley & Sons, Inc.