Quantitative spatiotemporal chemical profiling of individual lipid droplets by hyperspectral CARS microscopy in living human adipose-derived stem cells

Di Napoli, Claudia, Pope, Iestyn ORCID: https://orcid.org/0000-0002-4104-0389, Masia, Francesco ORCID: https://orcid.org/0000-0003-4958-410X, Langbein, Wolfgang ORCID: https://orcid.org/0000-0001-9786-1023, Watson, Pete ORCID: https://orcid.org/0000-0003-0250-7852 and Borri, Paola ORCID: https://orcid.org/0000-0002-7873-3314 2016. Quantitative spatiotemporal chemical profiling of individual lipid droplets by hyperspectral CARS microscopy in living human adipose-derived stem cells. Analytical Chemistry 88 (7) , pp. 3677-3685. 10.1021/acs.analchem.5b04468

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Abstract

There is increasing evidence showing that cytosolic lipid droplets, present in all eucaryotic cells, play a key role in many cellular functions. Yet their composition at the individual droplet level and how it evolves over time in living cells is essentially unknown due to the lack of suitable quantitative non-destructive measurement techniques. In this work we demonstrate the ability of label-free hyperspectral coherent anti-Stokes Raman scattering (CARS) microscopy, together with a quantitative image analysis algorithm developed by us, to quantify the lipid type and content in vol:vol concentration units of individual lipid droplets in living human adipose-derived stem cells during differentiation over 9 days in media supplemented with different fatty acids. Specifically, we investigated the addition of the poly-unsaturated linoleic and alpha-linolenic fatty acids into the normal differentiation medium (mostly containing mono-unsaturated fatty acids). We observe a heterogeneous uptake which is droplet-size dependent, time dependent, and lipid dependent. Cells grown in linoleic acid-supplemented medium show the largest distribution of lipid content across different droplets at all times during differentiation. When analyzing the average lipid content, we find that adding linoleic or alpha-linolenic fatty acids at day 0 results in uptake of the new lipid components with an exponential time constant of 22±2hr. Conversely, switching lipids at day 3 results in an exponential time constant of 60±5hr. These are unprecedented findings, exemplifying that the quantitative imaging method demonstrated here could open a radically new way of studying and understanding cytosolic lipid droplets in living cells.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Physics and Astronomy Biosciences
Publisher:	American Chemical Society
ISSN:	0003-2700
Date of First Compliant Deposit:	30 March 2016
Date of Acceptance:	2 March 2016
Last Modified:	17 Aug 2024 06:35
URI:	https://orca.cardiff.ac.uk/id/eprint/87656

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