| Abstract: | Optical clearing methods are widely used for three-dimensional biological information acquisition in the whole organ. However, the imaging quality of cleared tissues is often limited by ununiformed tissue clearing. By combining tissue clearing with mechanical sectioning based whole organ imaging system, we can reduce the influence of light scattering and absorption on the tissue to get isotropic and high resolution in both superficial and deep layers. However, it remains challenging for optical cleared biological tissue to maintain good sectioning property. Here, we developed a clearing method named M-CUBIC (machinable CUBIC), which combined a modified CUBIC method with PNAGA (poly-N-acryloyl glycinamide) hydrogel embedding to transparentize tissue while improving its sectioning property. With high-throughput light-sheet tomography platform (HLTP) and fluorescent micro-optical sectioning tomography (fMOST), we acquired continuous datasets with subcellular resolution from intact mouse brains for single neuron tracing, as well as the fine vascular structure of kidneys. This method can be used to acquire microstructures of multiple types of biological organs with subcellular resolutions, which can facilitate biological research. |
