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Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: potential for non-invasive glucose sensing in diabetes mellitus

Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: potential for non-invasive glucose sensing in diabetes mellitus
Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: potential for non-invasive glucose sensing in diabetes mellitus
The aim of this study was to test the hypothesis that glucose can be monitored non-invasively by measuring NAD(P)H-related fluorescence lifetime of cells in an in vitro cell culture model. Autofluorescence decay functions were measured in 3T3-L1 adipocytes by time-correlated single-photon counting (excitation 370nm, emission 420-480nm). Free NADH had a two-exponential decay but cell autofluorescence fitted best to a three-exponential decay. Addition of 30mM glucose caused a 29% increase in autofluorescence intensity, a significantly shortened mean lifetime (from 7.23 to 6.73ns), and an increase in the relative amplitude and fractional intensity of the short-lifetime component at the expense of the two longer-lifetime components. Similar effects were seen with rotenone, an agent that maximizes mitochondrial NADH. 3T3-L1 fibroblasts stained with the fluorescent mitochondrial marker, rhodamine 123 showed a 16% quenching of fluorescence intensity when exposed to 30mM glucose, and an increase in the relative amplitude and fractional intensity of the short lifetime at the expense of the longer lifetime component. We conclude that, though the effect size is relatively small, glucose can be measured non-invasively in cells by monitoring changes in the lifetimes of cell autofluorescence or of a dye marker of mitochondrial metabolism. Further investigation and development of fluorescence intensity and lifetime sensing is therefore indicated for possible non-invasive metabolic monitoring in human diabetes.
fluorescence, glucose, diabetes mellitus, time-resolved fluorescence, time-correlated single-photon counting, non-invasive monitoring
1011-1344
122-129
Evans, N.D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Gnudi, L.
9d236366-c26e-4076-a0d7-ce3f56f2d4c4
Rolinski, O.J.
af593e53-8906-49fa-b779-b7cf67c20f3b
Birch, D.J.S.
36e7897a-ea5f-4fb4-85ca-e7f728176f02
Pickup, J.C.
4c76692b-1c77-42fc-a758-aa7f0263f385
Evans, N.D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Gnudi, L.
9d236366-c26e-4076-a0d7-ce3f56f2d4c4
Rolinski, O.J.
af593e53-8906-49fa-b779-b7cf67c20f3b
Birch, D.J.S.
36e7897a-ea5f-4fb4-85ca-e7f728176f02
Pickup, J.C.
4c76692b-1c77-42fc-a758-aa7f0263f385

Evans, N.D., Gnudi, L., Rolinski, O.J., Birch, D.J.S. and Pickup, J.C. (2005) Glucose-dependent changes in NAD(P)H-related fluorescence lifetime of adipocytes and fibroblasts in vitro: potential for non-invasive glucose sensing in diabetes mellitus. Journal of Photochemistry and Photobiology B: Biology, 80 (2), 122-129. (doi:10.1016/j.jphotobiol.2005.04.001). (PMID:15908228)

Record type: Article

Abstract

The aim of this study was to test the hypothesis that glucose can be monitored non-invasively by measuring NAD(P)H-related fluorescence lifetime of cells in an in vitro cell culture model. Autofluorescence decay functions were measured in 3T3-L1 adipocytes by time-correlated single-photon counting (excitation 370nm, emission 420-480nm). Free NADH had a two-exponential decay but cell autofluorescence fitted best to a three-exponential decay. Addition of 30mM glucose caused a 29% increase in autofluorescence intensity, a significantly shortened mean lifetime (from 7.23 to 6.73ns), and an increase in the relative amplitude and fractional intensity of the short-lifetime component at the expense of the two longer-lifetime components. Similar effects were seen with rotenone, an agent that maximizes mitochondrial NADH. 3T3-L1 fibroblasts stained with the fluorescent mitochondrial marker, rhodamine 123 showed a 16% quenching of fluorescence intensity when exposed to 30mM glucose, and an increase in the relative amplitude and fractional intensity of the short lifetime at the expense of the longer lifetime component. We conclude that, though the effect size is relatively small, glucose can be measured non-invasively in cells by monitoring changes in the lifetimes of cell autofluorescence or of a dye marker of mitochondrial metabolism. Further investigation and development of fluorescence intensity and lifetime sensing is therefore indicated for possible non-invasive metabolic monitoring in human diabetes.

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More information

Published date: 1 August 2005
Keywords: fluorescence, glucose, diabetes mellitus, time-resolved fluorescence, time-correlated single-photon counting, non-invasive monitoring

Identifiers

Local EPrints ID: 176171
URI: http://eprints.soton.ac.uk/id/eprint/176171
ISSN: 1011-1344
PURE UUID: 227ff00b-0634-46a9-a45c-1cce0402a426
ORCID for N.D. Evans: ORCID iD orcid.org/0000-0002-3255-4388

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Date deposited: 08 Mar 2011 09:50
Last modified: 14 Mar 2024 02:56

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Contributors

Author: N.D. Evans ORCID iD
Author: L. Gnudi
Author: O.J. Rolinski
Author: D.J.S. Birch
Author: J.C. Pickup

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