SmartSPIM is an advanced light sheet microscope capable of imaging large tissue samples with high speed and resolution. The system was designed from the ground up, with no limitation based on commercially available microscope bodies. The unique illumination arm of the SmartSPIM creates a flat-top illumination profile laterally, and a focused beam axially. To achieve homogeneous axial point spread functions (PSFz) across the entire FOV, the illumination optics scan the beam axially while synchronized to the camera’s rolling shutter detection.
Custom designed imaging chamber and sample holders can accommodate tissues as small as organoids to those as big as an entire adult mouse brain. Due to the unique design of the sample holder, there are no points of contact between the tissue sample and the mounting hardware, thus allowing the user more freedom to mount a variety of sample types and sizes in different orientations.
Tunable imaging parameters allow for high speed volumetric image capture while maintaining high quality resolution. A dynamic, axially swept light sheet is synchronized with the rolling shutter of the camera for confocal-like detection with high homogeneity of signal.
20 fps acquisition allows imaging of a mouse brain hemisphere less than 30 minutes (at 1.8 µm x 1.8 µm x 4 µm voxel size), enabling the user to image 20 samples or more per week.
The SmartSPIM system has been designed from the ground up with user customization and maximum flexibility in mind. Users are not locked in to using proprietary optics and camera systems, nor are they limited to specific tissue clearing and labeling paradigms.
Entire hemisphere was stained in 1 day with SmartLabel using SYTO 16 green fluorescent nucleic acid stain and imaged using SmartSPIM with a custom LifeCanvas 3.6x objective. Acquisition time was less than 1 hour. The high and uniform axial resolution allows for clear delineation of cell bodies in XY and importantly in Z.
*Due to the formation of the light sheet, axial resolution is dependent on imaging rate. The high end of the range is representative of our standard fast acquisition rate.
SmartSPIM image of fluorescent protein expression in a subset of neurons. High-resolution axial spacing (4 µm) enables seamless display of original XY-dimensional data in orthogonal directions (XZ & YZ), providing for accurate identification of each cell body.
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