Multiphoton-Excitation Fluorescence Microscopy and Membranes

2006 ◽  
pp. 247-266
Author(s):  
Luis A. Bagatolli
Keyword(s):  
Fluorescence Microscopy ◽  
Multiphoton Excitation

scholarly journals Multiphoton excitation fluorescence microscopy in planar membrane systems

2010 ◽  
Vol 1798 (7) ◽  
pp. 1301-1308 ◽  
Author(s):  
Jonathan Brewer ◽  
Jorge Bernardino de la Serna ◽  
Kerstin Wagner ◽  
Luis A. Bagatolli
Keyword(s):  
Fluorescence Microscopy ◽  
Membrane Systems ◽  
Multiphoton Excitation ◽  
Planar Membrane

How Multiphoton Excitation can Illuminate Biophysics

1997 ◽  
Vol 3 (S2) ◽  
pp. 299-300
Author(s):  
W. W. Webb
Keyword(s):  
Fluorescence Microscopy ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Infrared Light ◽  
Focal Volume ◽  
Multiphoton Excitation ◽  
Fluorescence Excitation ◽  
Photon Excitation ◽  
Rate Of Absorption ◽  
Molecular Excitation

Multiphoton molecular excitation by the strongly focused femtosecond pulses of infrared light generated as an 80 MHZ pulse train by a mode locked laser provides intrinsic submicron three dimensional spatial resolution of fluorescence excitation and photochemistry for laser scanning fluorescence microscopy. Because two-photon excitation requires simultaneous (∼10-16 seconds), absorption of two-photons focused laser intensities of about 1022 photons/cm2s are required. Since the rate of absorption is proportional to the square of the intensity, excitation is limited to the focal volume and is negligible elsewhere along the double cone of the focused illumination. Therefore, out of focus photodamage and fluorescence are generally negligible and laser scanning fluorescence microscopy with multiphoton excitation is intrinsically three dimensionally resolved with no out of focus background. Since the appropriate wave lengths are infrared for multiphoton excitation of ultraviolet or visible absorbing molecules, out of focus photodamage is eliminated. This allows imaging of useful ultraviolet absorbing indicators, vital DNA stains and autofluorescence in living cells with minimal, but not necessarily negligible, photodamage.

High-resolution fluorescence microscopy employing a cyclic sequential multiphoton excitation

2010 ◽  
Author(s):  
Keisuke Isobe ◽  
Akira Suda ◽  
Hiroshi Hashimoto ◽  
Fumihiko Kannari ◽  
Hiroyuki Kawano ◽  
...  
Keyword(s):  
High Resolution ◽  
Fluorescence Microscopy ◽  
Multiphoton Excitation

scholarly journals Diamond Raman Laser and Yb Fiber Amplifier for In Vivo Multiphoton Fluorescence Microscopy

2021 ◽  
Author(s):  
Shaun A Engelmann ◽  
Annie Zhou ◽  
Ahmed M Hassan ◽  
Michael R Williamson ◽  
Jeremy W Jarrett ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Fluorescence Microscopy ◽  
Repetition Rate ◽  
Raman Laser ◽  
Fiber Amplifier ◽  
Multiphoton Excitation ◽  
Mouse Cerebral Cortex ◽  
Cost Efficient ◽  
Fast Image Acquisition

Here we introduce a fiber amplifier and a diamond Raman laser that output high powers (6.5 W, 1.3 W) at valuable wavelengths (1060 nm, 1250 nm) for multiphoton excitation of red-shifted fluorophores. These custom excitation sources are both simple to construct and cost-efficient in comparison to similar custom and commercial alternatives. Furthermore, they operate at a repetition rate (80 MHz) that allows fast image acquisition using resonant scanners. We demonstrate our system's compatibility with fast resonant scanning, the ability to acquire neuronal images, and the capability to image vasculature at deep locations (>1 mm) within the mouse cerebral cortex.

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