Tumor cell differentiation by label-free fluorescence microscopy

Abstract

Abstract. Autofluorescence spectra, images, and decay kinetics of U251-MG glioblastoma cells prior and subsequent to activation of tumor suppressor genes are compared. While phase contrast images and fluorescence intensity patterns of tumor (control) cells and less malignant cells are similar, differences can be deduced from autofluorescence spectra and decay kinetics. In particular, upon near UV excitation, the fluorescence ratio of the free and protein-bound coenzyme nicotinamid adenine dinucleotide depends on the state of malignancy and reflects different cytoplasmic (including lysosomal) and mitochondrial contributions. While larger numbers of fluorescence spectra are evaluated by principal component analysis, a multivariate data analysis method, additional information on cell metabolism is obtained from spectral imaging and fluorescence lifetime imaging microscopy.

SLICE: determining cell differentiation and lineage based on single cell entropy

“…Determining the differentiation/stemness states of individual cells enables prediction of the dynamic genetic networks regulating cellular activities during organogenesis, repair and disease. Traditionally, the status of cell differentiation or stemness is determined by the expression of known differentiation markers (1) or by morphological features of cells (4,5) (e.g. size and shape).…”

NAD(P)H and Flavin Autofluorescence Correlation with ATP in Rat Livers with Different Metabolic Steady‐State Conditions

“…The pyridine nucleotides NADH and NADPH, commonly referred to as NAD(P)H, and flavins are now more than ever at the basis of in vivo , in situ cell and tissue autofluorescence (AF) studies, with a continuously growing diagnostic value as endogenous biomarkers of normal and altered metabolism .…”

Label-free optical imaging of live cells

“…NADH is involved in many pathways, mainly related with redox reactions, and its high amount in mitochondria allows these organelles to be distinguished in the cell matrix in a non-invasive way (Dellinger et al, 1998;Ramey et al, 2007;Sikder et al, 2005) where the morphology of these organelles reflects the health of mammalian cells (Rodrigues et al, 2011). A fluorescence microscope equipped with a polychromator and an image intensifying detection unit can be used to measure the differences in auto-fluorescence spectra to study the decay kinetics of cells in different states of tumour malignancy, which is related to the amount of free and protein-bound NADH, reflecting different contributions of the fluorescent signal from the cytoplasm and mitochondria (Weber et al, 2012).…”