Nonlinear vibrational microscopy

Details Nonlinear vibrational microscopy Nonlinear vibrational microscopy

1999-04-19

2000-08-22

Evans, F. L.

Optics: measuring and testing

By dispersed light spectroscopy

With raman type light scattering

G01J 344, G01N 2165

Patent

active

061080810

ABSTRACT:
The present invention is a method and apparatus for microscopic vibrational imaging using coherent Anti-Stokes Raman Scattering or Sum Frequency Generation. Microscopic imaging with a vibrational spectroscopic contrast is achieved by generating signals in a nonlinear optical process and spatially resolved detection of the signals. The spatial resolution is attained by minimizing the spot size of the optical interrogation beams on the sample. Minimizing the spot size relies upon a. directing at least two substantially co-axial laser beams (interrogation beams) through a microscope objective providing a focal spot on the sample; b. collecting a signal beam together with a residual beam from the at least two co-axial laser beams after passing through the sample; c. removing the residual beam; and d. detecting the signal beam thereby creating said pixel. The method has significantly higher spatial resolution then IR microscopy and higher sensitivity than spontaneous Raman microscopy with much lower average excitation powers. CARS and SFG microscopy does not rely on the presence of fluorophores, but retains the resolution and three-dimensional sectioning capability of confocal and two-photon fluorescence microscopy. Complementary to these techniques, CARS and SFG microscopy provides a contrast mechanism based on vibrational spectroscopy. This vibrational contrast mechanism, combined with an unprecedented high sensitivity at a tolerable laser power level, provides a new approach for microscopic investigations of chemical and biological samples.

REFERENCES:
patent: 4077719 (1978-03-01), Barrett et al.
patent: 4405237 (1983-09-01), Manuccia et al.
patent: 4512660 (1985-04-01), Goldberg
Duncan et al, Optics Letters, vol. 7, No. 8, Aug. 1982, pp. 350-352.

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