Scanningless depth-resolved microscopy is achieved through
spatial-temporal focusing and has been demonstrated
previously. The advantage of this method is that a large area may be imaged without scanning resulting in higher
throughput of the imaging system. Because it is a widefield technique, the optical sectioning effect is considerably
poorer than with conventional spatial focusing two-photon microscopy. Here we propose wide-field two-photon
microscopy based on spatio-temporal focusing and employing background rejection based on the HiLo microscope
principle. We demonstrate the effects of applying HiLo microscopy to widefield temporally focused two-photon
microscopy.
The study of second harmonic generation (SHG) has been examined using a vectorial approach for both linearly and radially polarized beams. This approach is necessary for situations when the beam is tightly focused such as in a microscope. Using the vectorial approach, the result of including the y and z components of the electric field is that previously ignored 'cross-component' terms are now found to have an influence on the SHG polarization and the radiation patterns obtained. Since SHG is dependent on the susceptibility tensor, the inclusion of these 'cross-component' terms can help to identify structural changes in biological materials simply by studying the changes in the tensor via the SHG polarization. In particular, we calculate the second harmonic polarization induced in collagen for both linearly and radially polarized beams.
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