Signal Discrimination Between Fluorescent Proteins in Live Cells by Long-wavelength Optical Modulation.

ABSTRACT: Fluorescent proteins (FPs) have revolutionized molecular and cellular biology; yet, discrimination over cellular autofluorescence, spectral deconvolution, or detection at low concentrations remain challenging problems in many biological applications. By optically depopulating a photoinduced dark state with orange secondary laser co-excitation, the higher-energy green AcGFP fluorescence is dynamically increased. Modulating this secondary laser then modulates the higher-energy, collected fluorescence; enabling its selective detection by removing heterogeneous background from other FPs. Order-of-magnitude reduction in obscuring fluorophore background emission has been achieved in both fixed and live cells. This longwavelength modulation expands the dimensionality to discriminate FP emitters b

SUBMITTER: Jablonski AE 

PROVIDER: S-EPMC3570161 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature