|A hyperspectral stimulated emission depletion ("STED") microscope system
for high-resolution imaging of samples labeled with multiple fluorophores
(e.g., two to ten fluorophores). The hyperspectral STED microscope
includes a light source, optical systems configured for generating an
excitation light beam and a depletion light beam, optical systems
configured for focusing the excitation and depletion light beams on a
sample, and systems for collecting and processing data generated by
interaction of the excitation and depletion light beams with the sample.
Hyperspectral STED data may be analyzed using multivariate curve
resolution analysis techniques to deconvolute emission from the multiple
fluorophores. The hyperspectral STED microscope described herein can be
used for multi-color, subdiffraction imaging of samples (e.g., materials
and biological materials) and for analyzing a tissue by Forster Resonance
Energy Transfer ("FRET").