| Summary: | Without sophisticated data inversion algorithms, nonlinear optical microscopy can acquire
images at subcellular resolution and relatively large depth, with plausible endogenous
contrasts indicative of authentic biological and pathological states. Although independent
contrasts have been derived by sequentially imaging the same sample plane or volume under
different and often optimized excitation conditions, new laser source engineering with
inputs from key biomolecules surprisingly enable real-time simultaneous acquisition of
multiple endogenous molecular contrasts to segment a rich set of cellular and
extracellular components. Since this development allows simple single-beam single-shot
excitation and simultaneous multicontrast epidirected signal detection, the resulting
platform avoids perturbative sample pretreatments such as fluorescent labeling, mechanical
sectioning, scarce or interdependent contrast generation, constraints to the sample or
imaging geometry, and intraimaging motion artifacts that have limited in
vivo nonlinear optical molecular imaging.
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