Hyerpsectral imaging flow cytometry

Color carries a wealth of information in fluorescence microscopy.  Organelles, proteins and DNA sequences can all be independently fluorescently labeled and then resolved by the color of their fluorescence emission.

Hyperspectral imaging system often require multiple sequential images, each taken through a different color filter, to assemble the hyperspectral data cube.  This relatively large acquisition time prohibits their use on fast moving samples, like flow cytometry, where cells can move several millimeter per second.

Recently we have demonstrated a hyperspectral microscope that uses microfluidic fluid flow to mechanically scan cells over different regions of a linear variable filter, and therefore exploits the fluid’s motion to capture the hyperspectral image set.

Images and spectra of leukemia cells (HL60) stained with a membrane dye (green) and a DNA stain (yellow).

Individual organelles can be spatially and spectrally resolved, as shown above in the case of peroxisomes in HL60 cells.

Hyperspectral fluorescence microfluidic microscopy, Biomed. Opt. Exp. 4 1486-1493 (2013).