Shot-Noise-Limited Two-Color Stimulated Raman Scattering Microscopy with a Balanced Detection Scheme.
2020
Stimulated Raman
scattering (SRS) microscopy has been considered
a useful technique for investigating chemical components by selectively
targeting the vibration mode of chemical structures. Its practical
application to the observation of molecular structures and dynamics
in complicated biological environments requires broad spectral coverage
with both high resolution and a high signal-to-noise ratio. Here,
we demonstrate a two-color SRS microscopy employing a balanced detection
scheme and a spectral focusing method. Two different SRS signals are
generated with pump and Stokes laser pulse pairs in perpendicular
polarization, where each of them acts as an intensity reference for
the other, significantly reducing the background noise level close
to the shot-noise limit even with a fiber-based femtosecond laser
system. The high spectral resolution comparable to that of spontaneous
Raman scattering spectroscopy is achieved with the spectral focusing
method. The two-color SRS images are obtained for a mixture of polymer
beads and for the distributions of lipids and proteins in U2OS cells.
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