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Measuring two-photon microscopy ultrafast laser pulse duration at the sample plane using time-correlated single-photon counting
- Source :
- Journal of Biomedical Optics
- Publication Year :
- 2020
- Publisher :
- Society of Photo-Optical Instrumentation Engineers, 2020.
-
Abstract
- pTwo-photon microscopy (2PM) has revolutionized biomedical imaging by allowing thin optical sectioning in relatively thick biological specimens. Because dispersive microscope components in 2PM, such as objective lens, can alter temporal laser pulse width (typically being broader at the sample plane), for accurate measurements of two-photon absorption properties, it is important to characterize pulse duration at the sample plane. We present a simple modification to a two-photon microscope light path that allows for second-harmonic-generation-based interferometric autocorrelation measurements to characterize ultrafast laser pulse duration at the sample plane using time-correlated single-photon counting (TCSPC). We show that TCSPC can be used as a simple and versatile method to estimate the zero time delay step value between two adjacent ultrafast laser pulses for these measurements. To demonstrate the utility of this modification, we measured the Coherent Chameleon-Ultra II Ti:sapphire laser pulse width at the sample plane using a 10 × air, 40 × air, or 63 × water-immersion objective lens. At 950-nm two-photon excitation, the measured pulse width was 154 ± 32, 165 ± 13, and 218 ± 27 fs (italicn/italic = 6, mean ± standard deviation), respectively./p.
- Subjects :
- Paper
Optics and Photonics
Microscope
Materials science
Optical sectioning
Biomedical Engineering
Physics::Optics
01 natural sciences
law.invention
010309 optics
Biomaterials
Optics
Two-photon excitation microscopy
Special Section Celebrating Thirty Years of Multiphoton Microscopy in the Biomedical Sciences
Electricity
law
0103 physical sciences
Microscopy
Photons
Microscopy, Confocal
business.industry
interferometric autocorrelation
Lasers
Pulse duration
Signal Processing, Computer-Assisted
Laser
Atomic and Molecular Physics, and Optics
Photon counting
Electronic, Optical and Magnetic Materials
time-correlated single-photon counting
pulse duration
Interferometry
Microscopy, Fluorescence, Multiphoton
multiphoton microscopy
business
Ultrashort pulse
second-harmonic generation
Subjects
Details
- Language :
- English
- ISSN :
- 15602281 and 10833668
- Volume :
- 25
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Journal of Biomedical Optics
- Accession number :
- edsair.doi.dedup.....14c0e212aa8f913139d0fca5672fd303