New imaging method tracks cell metabolism faster with less light
Cornell researchers report a two-photon fluorescence polarization method that monitors single-cell metabolism faster and with less light than standard FLIM, per a Science Advances study.
The story in focus
Cornell researchers designed a two-photon fluorescence polarization ratiometric microscopy method to observe cell metabolism in real time with less light than fluorescence lifetime imaging, according to findings published Sept. 4 in Science Advances. The approach could speed screening of therapeutic treatments.
- The method splits polarized fluorescence into two detectors to gauge how fast NADH molecules rotate.
- Slower rotation, with highly polarized emission, may indicate NADH bound to proteins and energy generation.
- The technique is presented as faster and lower-light than fluorescence lifetime imaging for tracking metabolism over time.
What changed
The method, two-photon fluorescence polarization ratiometric microscopy, scans polarized focused light across cells and uses a polarizing beam splitter to send fluorescence to two detectors at once. That yields a measure of how quickly emitting molecules rotate, rather than the photon-timing measurement used in fluorescence lifetime imaging, or FLIM.
The researchers say the readout gives similar information to FLIM faster and with less light, reducing exposure stress on cells imaged repeatedly over extended periods.
Who it affects
The work targets cancer metabolism research and imaging of three-dimensional tissue systems such as organoids and model organisms. The authors suggest simpler, lower-cost measurement could let more researchers design experiments watching dynamic responses in engineered tissue.
Cancer metabolism is described as changing within minutes or hours, which the team says standard tools tracked poorly.
What is available
The findings were published Sept. 4 in Science Advances. The instrumentation is described as less complicated than FLIM, but interpreting the data required new analysis and calibration methods and numerous control experiments. No product or clinical test is reported.
Why not just use fluorescence lifetime imaging?
The researchers say FLIM requires cells to be imaged at multiple time points over long periods, and too much light exposure stresses them. Their polarization method is presented as faster and lower-light.
Sources & further reading
- Fluorescent imaging tracks metabolism of cells in real time | Cornell Chronicle - Fluorescent imaging tracks metabolism of cells in real time ↗news.cornell.edu
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