S-Space College of Natural Sciences (자연과학대학) Dept. of Physics and Astronomy (물리·천문학부) Physics (물리학전공) Journal Papers (저널논문_물리학전공)
Superresolution fluorescence microscopy for 3D reconstruction of thick samples
- Park, Sangjun; Kang, Wooyoung; Kwon, Yeong-Dae; Shim, Jaehoon; Kim, Siyong; Kaang, Bong-Kiun; Hohng, Sungchul
- Issue Date
- BioMed Central
- Molecular Brain, 11(1):17
- Line-scan confocal microscopy; DNA-PAINT; Superresolution microscopy; Single-molecule localization; microscopy; Three-dimensional reconstruction
Three-dimensional (3D) reconstruction of thick samples using superresolution fluorescence microscopy remains challenging due to high level of background noise and fast photobleaching of fluorescence probes. We develop superresolution fluorescence microscopy that can reconstruct 3D structures of thick samples with both high localization accuracy and no photobleaching problem. The background noise is reduced by optically sectioning the sample using line-scan confocal microscopy, and the photobleaching problem is overcome by using the DNA-PAINT (Point Accumulation for Imaging in Nanoscale Topography). As demonstrations, we take 3D superresolution images of microtubules of a whole cell, and two-color 3D images of microtubules and mitochondria. We also present superresolution images of chemical synapse of a mouse brain section at different z-positions ranging from 0 μm to 100 μm.