期刊
MICROSCOPY AND MICROANALYSIS
卷 8, 期 3, 页码 191-202出版社
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927602020147
关键词
demodulation; fluorescence imaging; laser scanning microscopy; LSM; MPE; multiphoton excitation; phase sensitive detector; PSD; signal processing; two-photon excitation; TPE
资金
- NCI NIH HHS [CA86283] Funding Source: Medline
- NIDDK NIH HHS [DK53434] Funding Source: Medline
Multiphoton laser scanning microscopy offers advantages in depth of penetration into intact samples over other optical sectioning techniques. To achieve these advantages it is necessary to detect the emitted light without spatial filtering. In this nondescanned (nonconfocal) approach, ambient room light can easily contaminate the signal, forcing experiments to be performed in absolute darkness. For multiphoton microscope systems employing mode-locked lasers, signal processing can be used to reduce such problems by taking advantage of the pulsed characteristics of such lasers. Specifically, by recovering fluorescence generated at the mode-locked frequency, interference from stray light and other ambient noise sources can be significantly reduced. This technology can be adapted to existing microscopes by inserting demodulation circuitry between the detector and data collection system. The improvement in signal-to-noise ratio afforded by this approach yields a more robust microscope system and opens the possibility of moving multiphoton microscopy from the research lab to more demanding settings, such as the clinic.
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