Label-free discrimination of actin and myosin muscle networks using nonresonant contribution of Multiplex-Coherent anti-Stokes Raman Scattering (M-CARS)
arxiv(2024)
摘要
We propose an original experimental strategy providing evidence for the first
label-free disclosure of muscle actin network based on the nonresonant
contribution of a multiplex CARS (M-CARS) microscopy setup. Thin actin
filaments are known for being located between thick myosin filaments that can
be detected in Second Harmonic Generation (SHG) with our M-CARS setup. The
first step of our strategy consists in defining spatial regions where thick
myosin filaments or thin actin filaments are expected. Then we present an
analysis based on two M-CARS spectra corresponding to these regions, which
outcome highlights differences in their relative nonresonant contribution. The
final step of the presented strategy is their exploitation on each pixel of the
M-CARS hyperspectral image, enabling label-free revelation of thin actin
filaments. This qualitative imaging represents a proof of principle for
highlighting and discriminating purposes in biological microscopy thanks to a
difference in the nonlinear properties of the related proteins. Besides, this
peculiar imaging technique results from a competition between several four-wave
mixing schemes. This paves the way for considering label-free imaging through a
competition involving several light-matter interaction effects, rather than
considering several of them singularly.
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