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优化单分子定位显微镜的成像速度和激发强度。

Optimizing imaging speed and excitation intensity for single-molecule localization microscopy.

机构信息

Cell Biology and Biophysics Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.

Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.

出版信息

Nat Methods. 2020 Sep;17(9):909-912. doi: 10.1038/s41592-020-0918-5. Epub 2020 Aug 17.

DOI:10.1038/s41592-020-0918-5
PMID:32807954
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7610360/
Abstract

High laser powers are common practice in single-molecule localization microscopy to speed up data acquisition. Here we systematically quantified how excitation intensity influences localization precision and labeling density, the two main factors determining data quality. We found a strong trade-off between imaging speed and quality and present optimized imaging protocols for high-throughput, multicolor and three-dimensional single-molecule localization microscopy with greatly improved resolution and effective labeling efficiency.

摘要

高激光功率在单分子定位显微镜中很常见，可加快数据采集速度。在这里，我们系统地量化了激发强度如何影响定位精度和标记密度，这两个主要因素决定了数据质量。我们发现成像速度和质量之间存在强烈的权衡，并提出了优化的成像方案，用于高通量、多色和三维单分子定位显微镜，具有大大提高的分辨率和有效的标记效率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/8a0a44626650/EMS118375-f002.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/e816a23200e7/EMS118375-f009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/aa57133a19f3/EMS118375-f010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/3f5047625292/EMS118375-f011.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/8183923bbd64/EMS118375-f012.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/9c943b79854c/EMS118375-f001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73d1/7610360/8a0a44626650/EMS118375-f002.jpg

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