Multiphoton microscope

Dedicated TCSPC electronics is used in all practical TCSPC-FLIM implementations [21, 22]. There are several issues that should be noted. First of all, the lifetime acquisition has to be synchronized with the scanning of the confocal or multiphoton microscope. To this end, the pixel clock and often the line and frame synchronization signals of the scanning microscope are used. 

Straub, M. and Hell, S. W. (1998). Fluorescence lifetime three-dimensional microscopy with picosecond precision using a multifocal multiphoton microscope. Appl. Phys. Lett. 73, 1769-71. 

Dong, C. Y., Yu, B., Kaplan, P. D., and So, P. T. C. 2004. Performances of high NA water and oil immersion ohjective in deep tissue, multiphoton microscopic imaging of excised human skin. Micr. Res. Tech. 63 81-86. 

Konig, K., Becker, T. W., Fischer, R, Riemaim, L, and Halbhuber, K. J. 1999. Pulse-length dependence of cellular response to intense near-infrared laser pulses in multiphoton microscopes. Opt. Lett. 24 113-15. 

Figure 4.11 A three-dimensional reconstruction of multiphoton microscope acquired image slices of a series of small vessels in the dorsal skinfold window chamber. Taken with a 40x water immersion objective and digitally zoomed to approximately 60x.

Multiphoton Microscope and Photophysical Technologies Probing Molecular Dynamics of Life... 

Laser-scanning microscopes can be classified by the way they excite and detect fluorescence in the sample. One-photon microscopes use a NUV or visible CW laser to excite the sample. Two-photon, or Multiphoton , microscopes use a femtosecond laser of high repetition rate. The fluorescence light can be detected by feeding it back through the scanner and through a confocal pinhole. The principle is termed confocal or descanned detection. A second detection method is to divert the fluorescence directly behind the microscope objective. The principle is termed direct or nondescaimed detection. 

With a Ti Sapphire laser or another high-repetition rate femtosecond laser, the sample can be excited by simultaneous multiphoton absorption [132, 164, 278, 282, 343, 471, 472]. For biological specimens, three-photon or higher-order excitation is rarely used. Nevertheless, such microscopes are normally called Multiphoton microscopes. 

Upright multiphoton microscope equipped with (a) at least two (ideally three to four) non-descanned PMT detectors (e.g.. Ultima V, Prairie... 

Konig K (2000) Laser tweezers and multiphoton microscopes in life sciences. Histochem Cell Biol 114 79-92... 

In Situ Characterization of Membrane Fouling and Cleaning Using a Multiphoton Microscope... 

The chemical cleaning of protein fouled membranes was also investigated using the multiphoton microscope. There is potential for further work in this area. 

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