Modular dipping light sheet microscopy

Light sheet microscopy has become a popular imaging modality for multi-dimensional imaging. Its unconventional orthogonal setup between the excitation and emission axis allows for light to optically section the sample reducing the amount of background signal when compared to epifluorescence and reducing the laser power deposited into the sample when compared to confocal imaging. As the optics are different from a traditional inverted microscope, traditional mounting methods cannot be used. Instead, different mounting methods were created to work around the setup and adapt to the sample. Furthermore, a plethora of light sheet iterations were designed around different sample mounting approached. We present a vertical dipping light sheet system with modular capabilities, allowing for different experimental and imaging conditions to be configured. The system features a variety of different imaging objectives including an immersion objective of cleared samples. It also has ample space around the sample area permitting different apparatus to be used around the sample. Finally, the modular approach enables us to experiment and change imaging and sample handling parameters for different requirements. One experimental system developed on the microscope is a magnification toggle system where the system can toggle between high magnification and low magnification by swapping tube lenses within the optical axis. This is coupled with the switching of excitation optics optimizing the sheet width with the field of view of the system. Such a system would be useful for locating regions of interest before switching to a higher magnification for volume acquisition.