Spatio-temporally resolved observation and control of neuron excitement mechanism using nonlinear laser microscopes

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1.An over-one-octave spanning (460-1060 nm) ultra-broadband optical pulse was generated by self-modulation effect. It was spectrally-phase-compensated and characterized to be a 2.8 fs, 1.5 cycle-Fourier-transform-limited pulse by the modified spectral phase interferometry for direct electric-field reconstruction. It is the shortest visible and infrared pulse whose spectral phase is well determined.2.A second-harmonic laser microscope was developed for 3-dimensional spatially-resolved observation of living cells. As an example, neurons of a rat were observed. Strong signals from nuclei and axons owing to inversion-symmetry breaking were detected. Spatial resolution was less than 1.5 μm.3.A coherent anti-Stokes Raman scattering laser microscope was developed using a femtosecond laser amplifier as a light source. For onion cells, two pulses with wavelengths of 800 and 1040 nm were employed to excite the C-H vibration mode of 2900 cm-1 and the corresponding spatially-resolved signal with a wavelength of 650 nm was detected. Similarly, two pulses with wavelengths of 800 and 1100 nm were employed to excite the N-H vibration mode of 3400 cm-1 and the corresponding signal with a wavelength of 630 nm was detected. From comparison between those signals, C-H-and N-H-rich domains in cells were determined.4.Macroscopic observation of hybridization and dehybridization for methylred-intercalated DNA was performed using second-harmonic laser microscope. While hybridized one has the inversion symmetry, dehybridized one does not. Significant signals were not obtained so far because of noise. Improvement is going on by introducing other azo-dyes with higher optical nonlinearity.