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Measurement of expansion factor and distortion for expansion microscopy using isolated renal glomeruli as landmarks |
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| Authors: | Chen Zhu Aidong Wang Lili Chen Liangsheng Guo Jiajia Ye Qilin Chen Qi Wang Guojia Yao Qin Xia Tianyu Cai Jiayun Guo Zhenyu Yang Zhenglong Sun Yuwei Xu Guoyuan Lu Zexin Zhang Jingyuan Cao Ying Liu Huizhong Xu |
| Affiliation: | 1. Institute for Advanced Study, Soochow University, Suzhou, China School of Optoelectronic Science and Engineering, Soochow University, Suzhou, China;2. The Second Affiliated Hospital of Soochow University, Suzhou, China;3. Institute for Advanced Study, Soochow University, Suzhou, China School of Physical Science and Technology, Soochow University, Suzhou, China;4. Children's Hospital of Chongqing Medical University, Chongqing, China;5. Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, Suzhou, China;6. The First Affiliated Hospital of Soochow University, Suzhou, China;7. Institute for Advanced Study, Soochow University, Suzhou, China College of Chemistry, Chemical Engineering and Materials Science, Soochow University, Suzhou, China;8. Department of Nephrology, Taizhou People's Hospital, the Fifth Affiliated Hospital of Nantong University, Taizhou, China;9. Institute of Pediatric Research, Children's Hospital of Soochow University, Suzhou, China;10. Institute for Advanced Study, Soochow University, Suzhou, China |
| Abstract: | Expansion microscopy has enabled super resolution imaging of biological samples. The accurate measurement of expansion factor and distortion typically requires locating and imaging the same region of interest in the sample before and after expansion, which is often time-consuming to achieve. Here we introduce a convenient method for relocation by utilizing isolated porcine glomeruli as landmarks during expansion. Following heat denaturation and proteinase K digestion protocols, the glomeruli exhibit expansion factor of 3.5 to 4 (only 7%-16% less expanded than the hydrogel), and 1% to 2% of relative distortion. Due to its appropriate size of 100 to 300 μm, the location of the glomerulus in the sample are visible to eyes, while its detailed shape only requires bright field microscopy. For expansion factors ranging from 3 to 10, the region in the vicinity of the glomerulus can be easily re-identified, and sometimes allows quantification of expansion factor and distortion under bright field without fluorescent labels. |
| Keywords: | expansion factor expansion microscopy glomerulus super resolution |