High-throughput, nonperturbing quantification of lipid droplets with digital holographic microscopy.

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Version: Final published version
License: CC BY 4.0
Serval ID
serval:BIB_AE3526302F91
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
High-throughput, nonperturbing quantification of lipid droplets with digital holographic microscopy.
Journal
Journal of lipid research
Author(s)
Campos V., Rappaz B., Kuttler F., Turcatti G., Naveiras O.
ISSN
1539-7262 (Electronic)
ISSN-L
0022-2275
Publication state
Published
Issued date
07/2018
Peer-reviewed
Oui
Volume
59
Number
7
Pages
1301-1310
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
In vitro differentiating adipocytes are sensitive to liquid manipulations and have the tendency to float. Assessing adipocyte differentiation using current microscopy techniques involves cell staining and washing, while using flow cytometry involves cell retrieval in suspension. These methods induce biases, are difficult to reproduce, and involve tedious optimizations. In this study, we present digital holographic microscopy (DHM) as a label-free, nonperturbing means to quantify lipid droplets in differentiating adipocytes in a robust medium- to high-throughput manner. Taking advantage of the high refractive index of lipid droplets, DHM can assess the production of intracellular lipid droplets by differences in phase shift in a quantitative manner. Adipocytic differentiation, combined with other morphological features including cell confluence and cell death, was tracked over 6 days in live OP9 mesenchymal stromal cells. We compared DHM with other currently available methods of lipid droplet quantification and demonstrated its robustness with modulators of adipocytic differentiation in a dose-responsive manner. This study suggests DHM as a novel marker-free nonperturbing method to study lipid droplet accumulation and may be envisioned for drug screens and mechanistic studies on adipocytic differentiation.
Keywords
Adipocytes/cytology, Adipocytes/metabolism, Cell Differentiation, Cell Line, Holography, Image Processing, Computer-Assisted, Lipid Droplets/metabolism, Microscopy, adipocyte, adipogenesis, label-free
Pubmed
Web of science
Open Access
Yes
Create date
29/07/2018 13:26
Last modification date
04/05/2021 6:36
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