Resonant-scanning dual-color STED microscopy with ultrafast photon counting: A concise guide
Methods, ISSN: 1046-2023, Vol: 88, Page: 48-56
2015
- 18Citations
- 47Captures
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Metrics Details
- Citations18
- Citation Indexes16
- 16
- CrossRef15
- Patent Family Citations2
- Patent Families2
- Captures47
- Readers47
- 47
Article Description
STED (stimulated emission depletion) is a popular super-resolution fluorescence microscopy technique. In this paper, we present a concise guide to building a resonant-scanning STED microscope with ultrafast photon-counting acquisition. The STED microscope has two channels, using a pulsed laser and a continuous-wave (CW) laser as the depletion laser source, respectively. The CW STED channel preforms time-gated detection to enhance optical resolution in this channel. We use a resonant mirror to attain high scanning speed and ultrafast photon counting acquisition to scan a large field of view, which help reduce photobleaching. We discuss some practical issues in building a STED microscope, including creating a hollow depletion beam profile, manipulating polarization, and monitoring optical aberration. We also demonstrate a STED image enhancement method using stationary wavelet expansion and image analysis methods to register objects and to quantify colocalization in STED microscopy.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S1046202315300062; http://dx.doi.org/10.1016/j.ymeth.2015.06.019; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84959461905&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/26123183; https://linkinghub.elsevier.com/retrieve/pii/S1046202315300062
Elsevier BV
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