Journal
OPTICS COMMUNICATIONS
Volume 312, Issue -, Pages 62-67Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.optcom.2013.09.022
Keywords
Sub-diffraction; Photobleaching; Confocal microscopy
Categories
Funding
- National Natural Science Foundation of China [61205160, 61378051, 61377013, 61335003]
- National Program on Key Basic Research Project [2013CB910200]
- Doctoral Fund of the Ministiy of Education of China [20110101120061, 20120101130006]
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We propose a single molecule localization method which takes advantage of stochastic photobleaching to improve the resolution of confocal fluorescence microscopy. By detecting the stochastic intensity loss of fluorophores, each fluorophore in the field can be localized. When all locations are known, a subdiffraction image can be retrieved through single molecule localization algorithms. A confocal scheme is used to record the bleaching process of the sample. Each fluorophore can be localized from the recorded streaming followed by image subtraction. Compared with other single molecule localization concepts such as stochastic optical reconstruction microscopy (STORM) and photoactivated localization microscopy (PALM), this method does not require a laser cycling equipment and the pixel size is no longer limited by the size of CCD. This technique works well with common fluorescent dyes and does not require the use of engineered photoactivatable proteins or photoswitchable synthetic dye pairs. (c) 2013 Elsevier B.V. All rights reserved.
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