Phasor approach to fluorescence lifetime microscopy distinguishes different metabolic states of germ cells in a live tissue

C Stringari, A Cinquin, O Cinquin… - Proceedings of the …, 2011 - National Acad Sciences
C Stringari, A Cinquin, O Cinquin, MA Digman, PJ Donovan, E Gratton
Proceedings of the national academy of sciences, 2011National Acad Sciences
We describe a label-free imaging method to monitor stem-cell metabolism that discriminates
different states of stem cells as they differentiate in living tissues. In this method we use
intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging
microscopy in conjunction with image segmentation, which we use to introduce the concept
of the cell phasor. In live tissues we are able to identify intrinsic fluorophores, such as
collagen, retinol, retinoic acid, porphyrin, flavins, and free and bound NADH. We have …
We describe a label-free imaging method to monitor stem-cell metabolism that discriminates different states of stem cells as they differentiate in living tissues. In this method we use intrinsic fluorescence biomarkers and the phasor approach to fluorescence lifetime imaging microscopy in conjunction with image segmentation, which we use to introduce the concept of the cell phasor. In live tissues we are able to identify intrinsic fluorophores, such as collagen, retinol, retinoic acid, porphyrin, flavins, and free and bound NADH. We have exploited the cell phasor approach to detect a trend in metabolite concentrations along the main axis of the Caenorhabditis elegans germ line. This trend is consistent with known changes in metabolic states during differentiation. The cell phasor approach to lifetime imaging provides a label-free, fit-free, and sensitive method to identify different metabolic states of cells during differentiation, to sense small changes in the redox state of cells, and may identify symmetric and asymmetric divisions and predict cell fate. Our method is a promising noninvasive optical tool for monitoring metabolic pathways during differentiation or disease progression, and for cell sorting in unlabeled tissues.
National Acad Sciences