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Superresolution fluorescence microscopy for 3D reconstruction of thick samples.

ABSTRACT: 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 ?m to 100 ?m.

SUBMITTER: Park S

PROVIDER: S-EPMC5856285 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Superresolution fluorescence microscopy for 3D reconstruction of thick samples.

Park Sangjun S Kang Wooyoung W Kwon Yeong-Dae YD Shim Jaehoon J Kim Siyong S Kaang Bong-Kiun BK Hohng Sungchul S

Molecular brain 20180315 1

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 photobleac ...[more]

PMID: 29544505

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