Two-photon microscopy

Two-photon excitation by femtosecond NIR laser pulses can be used to obtain clear images of tissue layers as deep as 1 mm [132, 278, 279, 344, 462, 495, 534]. The efficiency of two-photon excitation depends on the square of the power density. It therefore works with noticeable efficiency only in the focus of the laser beam. With a microscope lens of high numerical aperture a lateral resolution around 300 nm and a longitudinal resolution of about 1 pm is obtained. Two-photon laser scanning microscopy has therefore become a standard technique of tissue microscopy. Two-photon laser scanning can be combined with... [Pg.124]

Nonlinear optics, such as two-photon microscopy, have historically been used for evaluating fluorescence signals in cell and animal model studies. " Unlike conventional confocal microscopy, two-photon microscopy relies on the use of an ultrafast pulsed laser (such as a femtosecond Tirsapphire laser) to... [Pg.643]

Lindek St, Cremer Chr and Stelzer E H K 1996 Confocal theta fluorescence microscopy using two-photon absorption and annular apertures Optik 02 131-4... [Pg.1674]

Bhawalkar J D, Swiatkiewicz J, Pan S J, Samarabandu J K, Liou W S, He G S, Berezney R, Cheng P C and Prasad P N 1996 Three-dimensional laser scanning two-photon fluorescence confocal microscopy of polymer materials using a new, efficient upconverting fluorophore Scanning 18 562-6... [Pg.1675]

Two-photon excited fluorescence detection at the single-molecule level has been demonstrated for cliromophores in cryogenic solids [60], room-temperature surfaces [61], membranes [62] and liquids [63, 64 and 65]. Altliough multiphoton excited fluorescence has been embraced witli great entluisiasm as a teclmique for botli ordinary confocal microscopy and single-molecule detection, it is not a panacea in particular, photochemical degradation in multiphoton excitation may be more severe tlian witli ordinary linear excitation, probably due to absorjDtion of more tlian tire desired number of photons from tire intense laser pulse (e.g. triplet excited state absorjDtion) [61],... [Pg.2493]

Strickler J FI and Webb W W 1990 Two-photon excitation in laser scanning fluorescence microscopy Proc. SPIE 13948107-18... [Pg.2506]

Brakenhoff G J, Squier J, Norris T, Bliton A C, Wade M FI and Athey B 1996 Real-time two-photon confocal microscopy using a femtosecond, amplified Ti sapphire system J. Microscopy 181 253-9... [Pg.2506]

The first-generation dendrimer 51 was directly observed by transmission electron microscopy (TEM). The TEM image showed that the dimensions of individual molecules are about 50 A, which is consistent with the calculated one [36]. Third-order NLO measurements showed a significant enhancement of two-photon absorption upon proceeding from the constituent molecules to the dendritic complex [35]. [Pg.60]

A nano-light-source generated on the metallic nano-tip induces a variety of optical phenomena in a nano-volume. Hence, nano-analysis, nano-identification and nanoimaging are achieved by combining the near-field technique with many kinds of spectroscopy. The use of a metallic nano-tip applied to nanoscale spectroscopy, for example, Raman spectroscopy [9], two-photon fluorescence spectroscopy [13] and infrared absorption spectroscopy [14], was reported in 1999. We have incorporated Raman spectroscopy with tip-enhanced near-field microscopy for the direct observation of molecules. In this section, we will give a brief introduction to Raman spectroscopy and demonstrate our experimental nano-Raman spectroscopy and imaging results. Furthermore, we will describe the improvement of spatial resolution... [Pg.24]

Sanchez, E. J., Novotny, L. and Xie, X. S. (1999) Near-field fiuorescence microscopy based on two-photon excitation with metal tips. Phys. Rev. Lett., 82, 4014-4017. [Pg.37]

W. W. (1990) Two-photon laser scanning fluorescence microscopy. Science, 248, 73-76. [Pg.37]

Williams RM, Piston DW, Webb WW (1994) Two-photon molecular excitation provides intrinsic 3-dimensional resolution for laser-based microscopy and microphotochemistry. FASEB J 8 804-813... [Pg.143]

So PT, Dong CY, Masters BR, Berland KM (2000) Two-photon excitation fluorescence microscopy. Annu Rev Biomed Eng 2 399-429... [Pg.143]

Piston DW (1999) Imaging living cells and tissues by two-photon excitation microscopy. Trends Cell Biol 9 66—69... [Pg.143]

Rubart M (2004) Two-photon microscopy of cells and tissue. Circ Res 95 1154—1166... [Pg.143]

Diaspro A, Chirico G, Federici F, Cannone F, Beretta S, Robello M (2001) Two-photon microscopy and spectroscopy based on a compact confocal scanning head. J Biomed Opt 6 300-310... [Pg.143]

Scherschel JA, Rubart M (2008) Cardiovascular imaging using two-photon microscopy. Microsc Microanal 14 492-506... [Pg.143]

Mondal PP, Diaspro A (2007) Reduction of higher-order photobleaching in two-photon excitation microscopy. Phys Rev E 75 6. doi 10.1103/PhysRevE. 1175.061904... [Pg.383]

Patterson GH, Piston DW (2000) Photobleaching in two-photon excitation microscopy. Biophys J 78 2159-2162... [Pg.383]

Push-push/pull-pull chromophores 118 and 119 have exhibited material properties to show their prospect for several applications particularly in the fields of two-photon microscopy and optical power limitation <1999CC2055, 2002SM17>. [Pg.705]

The conclusion from Grothwahl et al. [17] may also have some caveats. Since evidence from in vivo longitudinal two-photon microscopy... [Pg.319]

Lifetime imaging can be implemented both in wide field and in scanning microscopes such as confocal microscopes and two-photon excitation microscopes. The most common implementations in time-domain fluorescence lifetime imaging microscopy (FLIM) are based on TCSPC [8, 9] and time-gating (TG) [2, 10],... [Pg.110]

Buist, A. H., Muller, M., Squier, J. and Brakenhofif, G. J. (1998). Real time two-photon absorption microscopy using multi point excitation. J. Microsc. 192, 217-26. [Pg.177]

As mentioned above, spectral imaging microscopy is a form of multidimensional fluorescent microscopy where a fluorescent emission spectrum is acquired at each coordinate location in the sample. This mode of imaging has been implemented for wide field, confocal, and two-photon laser scanning microscopy, and several excellent... [Pg.363]

Tada, J., Kono, T., Suda, A., Mizuno, H., Miyawaki, A., Midorikawa, K. and Kannari, F. (2007). Adaptively controlled supercontinuum pulse from a micro structure fiber for two-photon excited fluorescence microscopy. Appl. Opt. 46, 3023-30. [Pg.515]

Bowman R.D., Kneas K.A., Demas J.N., Periasamy A., Conventional, Confocal And Two-Photon Fluorescence Microscopy Investigations of Polymer-Supported Oxygen Sensors, J. Microscopy 2003 211 112-120. [Pg.114]

Bird DK, Eliceiri KW, Fan CH, White JG (2004) Simultaneous two-photon spectral and lifetime fluorescence microscopy. Appl Opt 43 5173-82... [Pg.132]

For samples thicker than the depth of field, the images are blurred by out-of-focus fluorescence. Corrections using a computer are possible, but other techniques are generally preferred such as confocal microscopy and two-photon excitation microscopy. It is possible to overcome the optical diffraction limit in near-field scanning optical microscopy (NSOM). [Pg.354]

Big Chemical Encyclopedia

Chemical substances, components, reactions, process design ...