Abstract

In fluorescence microscopy, 3D image data is typically formed by acquiring a series of 2D images while changing the focal plane. This process is slow as it involves mechanically moving the sample or the objective, and as such forms a bottle neck for volumetric imaging. Here I present two applications of remote focusing that can significantly speed up volumetric imaging, both in two photon raster scanning microscopy and in light-sheet fluorescence microscopy.

I will further present a novel way to form projections of microscopic 3D objects under varying viewing angles. This method allows capturing views of a 3D sample at ~hundredfold increased speed compared to conventional 3D image acquisition. This can be valuable if the object is sufficiently sparse, and one does not need the full 3D information. But I will also show that it is possible to obtain three-dimensional reconstructions from a few rapidly acquired projections.

All three methods will be carefully characterized for their optical performance and examples of rapid biomedical imaging will be given.