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| CARS Techniques |
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Detection Schemes for CARS:
The signal is detected in the phase-matched direction and the signal is selected by a set of spectral filters. The F-CARS signals are generally very strong and can sometimes be observed by the naked eye.
The forward CARS signal is accompanied by a strong nonresonant background which may overshadow weak signals that are of interest. When detected in the backward direction, the nonresonant signal from the solvent (water) is completely eliminated. E-CARS is particularly sensitive to objects in focus that are smaller than the optical wavelength. When the sample is highly scattering, the forward propagating CARS signal can be backscattered, giving rise to a strong epi-signal .
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[Left] CARS Tissue image of small adipocytes of the subcutaneous layer from mouse ear tissue. The image was taken at a Raman shift of 2845 cm-1 to address the CH2 stretching frequency.
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By taking advantage of the Raman depolarization ratio of certain modes, the resonant signal can be separated from the nonresonant background when polarization sensitive detection is employed. P-CARS significantly enhances the image quality by eliminating the nonresonant background completely.
By employing a femtosecond time-scale delay between the excitation and probe pulses, time-resolved CARS allows for the complete suppression of nonresonant signals.
By interfering the CARS signal from a sample with a local oscillator CARS signal it is possible to selectively image the real and imaginary part of the third-order nonlinear susceptibility. iCARS enables rapid, amplified Raman imaging that is free of the nonresonant background.
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[Left] Comparison of heterodyne CARS with non-interferometric CARS imaging of live NIH 3T3 cells. The non-interferometric image of the cell in (a) is taken at the peak of the symmetric CH 2 vibration (2845 cm -1 ), with the measured imaginary and real response given in (b) and (c), respectively. The non-interferometric image of the cell in (d) is taken on the blue side of the CH-stretching band at 2950 cm -1 , with the isolated imaginary and real response shown in (e) and (f), respectively. Figures (g), (h) and (i) represent the non-interferometric, imaginary response and real response images off-resonance at 2086 cm -1 , respectively. Image dimensions are 35 x 40 ì m 2
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CARS Microspectroscopy Techniques: In addition to imaging, CARS spectroscopy can be carried out in the microscope. We have developed the following tools:
A picosecond laser is combined with a femtosecond laser to cover a broad range of vibrational frequencies. The CARS signal is collected and analyzed on a spectrometer. Chemical details in microscopic volumes can be quickly characterized with this method.
Analogous to fluorescence correlation spectroscopy, the autocorrelation of time-resolved CARS signals reflects rapid diffusion dynamics of small Raman scatterers in focus without the need for labeling.
Essentially a combination of interferometric CARS microscopy and multiplex CARS, interferometric CARS Microspectroscopy enables detection of the real and imaginary components of the CARS signal.
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