High-speed light-sheet fluorescent microscopy with an adaptive three-dimensional region-of-signal identification method

Abstract

Light-sheet fluorescent microscopy (LSFM) is a crucial 3D imaging technique for tissues with advantages in high 3D spatial resolution, effective optical sectioning, and quick imaging. Prior to imaging, the sample's imaging depth, imaging region, and region of interest (ROI) division should all be established in accordance with the sample's outermost dimensions. However, because biological samples are inherently irregular a traditional imaging flow that images ROI by ROI, slice by slice, would waste a great deal of time and data storage space by additionally recording non-signal regions. In order to solve this problem, we create a high-speed LSFM with an adaptive three-dimensional region-of-signal identification method (RSI). We immediately perform crude imaging for each ROI before the software analyzes the longitudinal distribution and, ultimately, uses dimensionality reduction, binarization, filtering, and redundancy expansion to acquire the signal region result. By imaging two mouse spinal cord segments with a total of 170 ROIs and 180 ROIs on two channels, respectively, the high-speed imaging capability is validated. Our method reduced the overall imaging time and raw data size by more than treble and 14 times, respectively, without losing signal. With a programmable imaging range based on the real signal region, this method offers an efficient tool for flexible light-sheet microscopy. It can also be applied to other microscopy techniques to speed up imaging.