Efficient and Accurate Synapse Detection With Selective Structured Illumination Microscopy on the Putative Regions of Interest of Ultrathin Serial Sections

Frontiers in Neuroanatomy

Jinseop S. Kim¹  Gyeong Tae Kim²  Jong-Cheol Rah³  Sangkyu Bahn⁴  Joon Ho Choi⁴  Nari Kim⁴ 

[1] Department of Biological Sciences, Sungkyunkwan University, Suwon, South Korea;Department of Biomedical Engineering, Ulsan National Institute of Science and Technology, Ulsan, South Korea;Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science and Technology, Daegu, South Korea;Korea Brain Research Institute, Daegu, South Korea;

关键词: structured illumination microscopy;    array tomography;    synapse location;    posterior medial nucleus;    barrel cortex;   

DOI  :  10.3389/fnana.2021.759816

来源: DOAJ

【 摘 要 】

Critical determinants of synaptic functions include subcellular locations, input sources, and specific molecular characteristics. However, there is not yet a reliable and efficient method that can detect synapses. Electron microscopy is a gold-standard method to detect synapses due to its exceedingly high spatial resolution. However, it requires laborious and time-consuming sample preparation and lengthy imaging time with limited labeling methods. Recent advances in various fluorescence microscopy methods have highlighted fluorescence microscopy as a substitute for electron microscopy in reliable synapse detection in a large volume of neural circuits. In particular, array tomography has been verified as a useful tool for neural circuit reconstruction. To further improve array tomography, we developed a novel imaging method, called “structured illumination microscopy on the putative region of interest on ultrathin sections”, which enables efficient and accurate detection of synapses-of-interest. Briefly, based on low-magnification conventional fluorescence microscopy images, synapse candidacy was determined. Subsequently, the coordinates of the regions with candidate synapses were imaged using super-resolution structured illumination microscopy. Using this system, synapses from the high-order thalamic nucleus, the posterior medial nucleus in the barrel cortex were rapidly and accurately imaged.

【 授权许可】

Unknown