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Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential.

PloS one | November 11, 2012 |

Stringari, Chiara | Nourse, Jamison L | Flanagan, Lisa A | Gratton, Enrico
Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Abstract

In the stem cell field there is a lack of non invasive and fast methods to identify stem cell's metabolic state, differentiation state and cell-lineage ... des a novel, and quantitative measure of stem cell potential and a label-free and non-invasive means to identify neuron- or glial- biased progenitors.

In the stem cell field there is a lack of non invasive and fast methods to identify stem cell's metabolic state, differentiation state and cell-lineage commitment. Here we describe a label-free method that uses NADH as an intrinsic biomarker and the Phasor approach to Fluorescence Lifetime microscopy to measure the metabolic fingerprint of cells. We show that different metabolic states are related to different cell differentiation stages and to stem cell bias to neuronal and glial fate, prior the expression of lineage markers. Our data demonstrate that the NADH FLIM signature distinguishes non-invasively neurons from undifferentiated neural progenitor and stem cells (NPSCs) at two different developmental stages (E12 and E16). NPSCs follow a metabolic trajectory from a glycolytic phenotype to an oxidative phosphorylation phenotype through different stages of differentiation. NSPCs are characterized by high free/bound NADH ratio, while differentiated neurons are characterized by low free/bound NADH ratio. We demonstrate that the metabolic signature of NPSCs correlates with their differentiation potential, showing that neuronal progenitors and glial progenitors have a different free/bound NADH ratio. Reducing conditions in NPSCs correlates with their neurogenic potential, while oxidative conditions correlate with glial potential. For the first time we show that FLIM NADH metabolic fingerprint provides a novel, and quantitative measure of stem cell potential and a label-free and non-invasive means to identify neuron- or glial- biased progenitors.

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Fig 1. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g001
Fig 1. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 1

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Fig 2. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g002
Fig 2. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 2

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Fig 3. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g003
Fig 3. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 3

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Fig 4. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g004
Fig 4. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 4

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Fig 5. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g005
Fig 5. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 5

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Fig 6. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g006
Fig 6. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 6

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Fig 7. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.
Pone.0048014.g007
Fig 7. Stringari, Nourse, et al. "Phasor fluorescence lifetime microscopy of free and protein-bound NADH reveals neural stem cell differentiation potential." PloS one 7.11 (2012): e48014. Web.

Fig 7

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