Apparatus and method for multiplexed one-photon and nonlinear microscopy and method for biological tissue alignment

Abstract

We combine linear and nonlinear microscopy modalities. This approach is capable of imaging a sample using the two modalities concurrently, generating time sequences of images of the sample having different properties characteristic of each of the modalities. We can separate light emitted from the sample, generated by either modality, even if the spectrum of light is identical for each modality. The nonlinear microscopy modality allows one to create high resolution volumetric images of the sample. The concurrent imaging of the sample using linear and nonlinear microscopy modalities enables one to correlate a spatial structure and temporal dynamics of the sample acquired in each modality allowing one to create a mapping between images across the modalities. The created mapping can be used to match images of a tissue sample acquired using linear modality in a living organism with high resolution, two- or three-dimensional images obtained using nonlinear microscopy modality.

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