Optimal integration of wide field illumination and holographic optical tweezers for multimodal microscopy with ultimate flexibility and versatility.

Abstract

We introduce one-of-a-kind optical microscope that we have developed through optimized integration of wide-field and focused-light microscopies. This new instrument has accomplished operation of the same laser for both wide field illumination and holographic focused beam illumination interchangeably or simultaneously in a way scalable to multiple lasers. We have demonstrated its powerful capability by simultaneously carrying out Epi-fluorescence, total internal reflection fluorescence microscopy, selective plane illumination microscopy, and holographic optical tweezers with five lasers. Our instrument and the optical design will provide researchers across diverse fields, cell-biology and biophysics in particular, with a practical guidance to build an all-around multimodal microscope that will further inspire the development of novel hybrid microscopy experiments.