The number of color channels that can be concurrently probed in fluorescence microscopy is severely limited by the broad fluorescence spectral width. Spectral imaging offers potential solutions, yet typical approaches to disperse the local emission spectra notably impede the attainable throughput.
UC Berkeley researchers have discovered methods and systems for simultaneously imaging up to 6 subcellular targets, labeled by common fluorophores of substantial spectral overlap, in live cells at low (~1%) crosstalks and high temporal resolutions (down to ~10 ms), using a single, fixed fluorescence emission detection band.
spectral microscopy of highly multiplexed fluorescence imaging
The ability to quantify the abundances of different fluorophores in the
same sample through unmixing the excitation spectra enables us to devise
quantitative imaging schemes for both bi-state and FRET fluorescent biosensors in live cells.
Achieve high sensitivities and spatiotemporal resolutions