NADH distribution in live progenitor stem cells by phasor-fluorescence lifetime image microscopy

Belinda K. Wright; Laura M. Andrews; Julie Markham; Mark R. Jones; Chiara Stringari; Michelle A. Digman; Enrico Gratton

doi:10.1016/j.bpj.2012.05.038

NADH distribution in live progenitor stem cells by phasor-fluorescence lifetime image microscopy

Belinda K. Wright
, Laura M. Andrews
, Julie Markham
, Mark R. Jones
, Chiara Stringari
, Michelle A. Digman
, Enrico Gratton

Western Sydney University
Long Beach VA and University of California

Research output: Contribution to journal › Article › peer-review

Abstract

NADH is a naturally fluorescent metabolite associated with cellular respiration. Exploiting the different fluorescence lifetime of free and bound NADH has the potential to quantify the relative amount of bound and free NADH, enhancing understanding of cellular processes including apoptosis, cancer pathology, and enzyme kinetics. We use the phasor- fluorescence lifetime image microscopy approach to spatially map NADH in both the free and bound forms of live undifferentiated and differentiated myoblast cells. The phasor approach graphically depicts the change in lifetime at a pixel level without the requirement for fitting the decay. Comparison of the spatial distribution of NADH in the nucleus of cells induced to differentiate through serum starvation and undifferentiated cells show differing distributions of bound and free NADH. Undifferentiated cells displayed a short lifetime indicative of free NADH in the nucleus and a longer lifetime attributed to the presence of bound NADH outside of the nucleus. Differentiating cells displayed redistribution of free NADH with decreased relative concentration of free NADH within the nucleus whereas the majority of NADH was found in the cytoplasm.

Original language	English
Pages (from-to)	L7-L9
Journal	Biophysical Journal
Volume	103
Issue number	1
DOIs	https://doi.org/10.1016/j.bpj.2012.05.038
Publication status	Published - 3 Jul 2012
Externally published	Yes

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This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

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10.1016/j.bpj.2012.05.038

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title = "NADH distribution in live progenitor stem cells by phasor-fluorescence lifetime image microscopy",

abstract = "NADH is a naturally fluorescent metabolite associated with cellular respiration. Exploiting the different fluorescence lifetime of free and bound NADH has the potential to quantify the relative amount of bound and free NADH, enhancing understanding of cellular processes including apoptosis, cancer pathology, and enzyme kinetics. We use the phasor- fluorescence lifetime image microscopy approach to spatially map NADH in both the free and bound forms of live undifferentiated and differentiated myoblast cells. The phasor approach graphically depicts the change in lifetime at a pixel level without the requirement for fitting the decay. Comparison of the spatial distribution of NADH in the nucleus of cells induced to differentiate through serum starvation and undifferentiated cells show differing distributions of bound and free NADH. Undifferentiated cells displayed a short lifetime indicative of free NADH in the nucleus and a longer lifetime attributed to the presence of bound NADH outside of the nucleus. Differentiating cells displayed redistribution of free NADH with decreased relative concentration of free NADH within the nucleus whereas the majority of NADH was found in the cytoplasm.",

author = "Wright, \{Belinda K.\} and Andrews, \{Laura M.\} and Julie Markham and Jones, \{Mark R.\} and Chiara Stringari and Digman, \{Michelle A.\} and Enrico Gratton",

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doi = "10.1016/j.bpj.2012.05.038",

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T1 - NADH distribution in live progenitor stem cells by phasor-fluorescence lifetime image microscopy

AU - Wright, Belinda K.

AU - Andrews, Laura M.

AU - Markham, Julie

AU - Jones, Mark R.

AU - Stringari, Chiara

AU - Digman, Michelle A.

AU - Gratton, Enrico

PY - 2012/7/3

Y1 - 2012/7/3

N2 - NADH is a naturally fluorescent metabolite associated with cellular respiration. Exploiting the different fluorescence lifetime of free and bound NADH has the potential to quantify the relative amount of bound and free NADH, enhancing understanding of cellular processes including apoptosis, cancer pathology, and enzyme kinetics. We use the phasor- fluorescence lifetime image microscopy approach to spatially map NADH in both the free and bound forms of live undifferentiated and differentiated myoblast cells. The phasor approach graphically depicts the change in lifetime at a pixel level without the requirement for fitting the decay. Comparison of the spatial distribution of NADH in the nucleus of cells induced to differentiate through serum starvation and undifferentiated cells show differing distributions of bound and free NADH. Undifferentiated cells displayed a short lifetime indicative of free NADH in the nucleus and a longer lifetime attributed to the presence of bound NADH outside of the nucleus. Differentiating cells displayed redistribution of free NADH with decreased relative concentration of free NADH within the nucleus whereas the majority of NADH was found in the cytoplasm.

AB - NADH is a naturally fluorescent metabolite associated with cellular respiration. Exploiting the different fluorescence lifetime of free and bound NADH has the potential to quantify the relative amount of bound and free NADH, enhancing understanding of cellular processes including apoptosis, cancer pathology, and enzyme kinetics. We use the phasor- fluorescence lifetime image microscopy approach to spatially map NADH in both the free and bound forms of live undifferentiated and differentiated myoblast cells. The phasor approach graphically depicts the change in lifetime at a pixel level without the requirement for fitting the decay. Comparison of the spatial distribution of NADH in the nucleus of cells induced to differentiate through serum starvation and undifferentiated cells show differing distributions of bound and free NADH. Undifferentiated cells displayed a short lifetime indicative of free NADH in the nucleus and a longer lifetime attributed to the presence of bound NADH outside of the nucleus. Differentiating cells displayed redistribution of free NADH with decreased relative concentration of free NADH within the nucleus whereas the majority of NADH was found in the cytoplasm.

UR - https://www.scopus.com/pages/publications/84863441183

U2 - 10.1016/j.bpj.2012.05.038

DO - 10.1016/j.bpj.2012.05.038

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AN - SCOPUS:84863441183

SN - 0006-3495

VL - 103

SP - L7-L9

JO - Biophysical Journal

JF - Biophysical Journal

IS - 1

ER -

NADH distribution in live progenitor stem cells by phasor-fluorescence lifetime image microscopy

Abstract

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