Second harmonic generation from thick leaves using the two-photon laser scanning microscope

The novelty of this work is to establish new compact technique in which one can use the SH imaging to investigate the true architecture of the sensitive samples, the unknown samples and the samples which is not producing auto-fluorescence. Moreover, investigation of new or unknown samples needs a long time for looking at details of the sample. Thereby the sample will be exposed for long time to the laser radiation that will cause photobleaching and photodamage. Since the SHG does not undergo photobleaching and photodamage this will be the promising technique for investigating the sensitive and new unknown samples. Then one can move to acquire fluorescence images after complete investigation of the true architecture of the sample. The other advantage of SHG is that it has the ability to image highly ordered structural proteins without any exogenous labels. The SHG is an intrinsic and a coherent process. Imaging of intrinsic compounds avoids the complications of slicing and labeling, and samples can be investigated under physiological conditions.