Phasor Approach to Fluorescence Microscopy Evaluates Cell Metabolism in vivo
Tech ID: 21811 / UC Case 2010-805-0
Researchers at the University of California, Irvine have developed a novel, label-free imaging and evalution method that enables users to track cell metabolism in vivo.
The technique is a novel phasor approach to Fluorescence Lifetime Imaging Microscopy (FLIM), a multi-photon microscopy technique that excites cells and then detects their fluorescence activity over time. In this approach, the data from these images is transformed mathematically into a phasor representation. The subsequent analysis identifies, locates, and calculates the concentration of important metabolic cell components, such as: collagen, FAD, free and bound NADH, retinol, and retinoic acid.
Overall, this novel method provides a straightforward and quantitative interpretation of the physiological processes occurring in tissues. It enables users to visualize cellular metabolism and retinoid gradients, distinguish between the unique metabolic states of cells, and map their level of differentiation.
In the past, immunostaining and metabolic assays were used to analyze cell metabolism. However, these methods are time consuming, invasive, and ultimately render the cells unviable.
To address these issues, non-invasive optical techniques have been developed recently that utilize multi-photon microscopy, taking advantage of the intrinsic auto-fluorescence of cells and tissues. In Fluorescence Lifetime Imaging Microscopy (FLIM), living tissue is excited by two-photon microscopy and the subsequent auto-fluorescence is detected and monitored over time. Currently, however, these techniques are lacking in their high variability and limited ability to assign fluorescence to and discriminate between specific tissue components.
This invention is a novel phasor approach to FLIM. The data from fluorescence lifetime images is transformed mathematically into a phasor representation. An analysis is then performed to locate, identify, and quantify the intrinsic fluorescent chemical species present. The approach has so far proven capable of identifying collagen, FAD, free and bound NADH, retinol and retinoic acid within living tissue. The relative concentrations of these tissue components are then calculated using a graphical analysis of the images.
The proposed phasor approach is intended for use in biology, biophotonics, and biomedical research to track in vivo metabolic changes associated with stem cell differentiation, cell carcinogenesis, apoptosis, and necrosis in tissues.
This approach might also be of interest to label-free cell sorting and high throughput screening for drug discovery, cell replacement therapies, and tissue engineering.
This method has been tested using living tissue.
|United States Of America||Published Application||20120276578||11/01/2012||2010-805|
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