Photoactivatable AMPA for the study of glutamatergic neuronal transmission using two-photon excitation

Author(s):  
Davide Deodato ◽  
Naeem Asad ◽  
Timothy M. Dore

TMP-CyHQ-AMPA is a photoactivatable form of a glutamate receptor agonist that has high sensitivity to 2-photon excitation. It can be used to study glutamatergic transmission with exceptional spatial-temporal resolution in complex tissue preparations.

2015 ◽  
Vol 6 (4) ◽  
pp. 2419-2426 ◽  
Author(s):  
Karolina A. Korzycka ◽  
Philip M. Bennett ◽  
Eduardo Jose Cueto-Diaz ◽  
Geoffrey Wicks ◽  
Mikhail Drobizhev ◽  
...  

We present a modular approach to photo-labile protecting groups based on photoinduced electron transfer, providing high sensitivity to two-photon excitation.


2004 ◽  
Author(s):  
Giuseppe Chirico ◽  
Gabriele Malengo ◽  
Roberto Milani ◽  
Fabio Cannone ◽  
Silke Krol ◽  
...  

Author(s):  
David W. Piston ◽  
Brian D. Bennett ◽  
Robert G. Summers

Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. Two-photon excitation arises from the simultaneous absorption of two photons in a single quantitized event whose probability is proportional to the square of the instantaneous intensity. For example, two red photons can cause the transition to an excited electronic state normally reached by absorption in the ultraviolet. In practice, two-photon excitation is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10-5 maintains the average input power on the order of 10 mW, only slightly greater than the power normally used in confocal microscopy.


Author(s):  
David W. Piston

Two-photon excitation fluorescence microscopy provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging. Two-photon excitation arises from the simultaneous absorption of two photons in a single quantitized event whose probability is proportional to the square of the instantaneous intensity. For example, two red photons can cause the transition to an excited electronic state normally reached by absorption in the ultraviolet. In our fluorescence experiments, the final excited state is the same singlet state that is populated during a conventional fluorescence experiment. Thus, the fluorophore exhibits the same emission properties (e.g. wavelength shifts, environmental sensitivity) used in typical biological microscopy studies. In practice, two-photon excitation is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 maintains the average input power on the order of 10 mW, only slightly greater than the power normally used in confocal microscopy.


Bioimaging ◽  
1995 ◽  
Vol 3 (2) ◽  
pp. 70-75 ◽  
Author(s):  
Pekka E Hänninen ◽  
Martin Schrader ◽  
Erkki Soini ◽  
Stefan W Hell

1997 ◽  
Vol 3 (S2) ◽  
pp. 305-306
Author(s):  
David W. Piston

Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. It provides three-dimensional resolution and eliminates background equivalent to an ideal confocal microscope without requiring a confocal spatial filter, whose absence enhances fluorescence collection efficiency. This results in inherent submicron optical sectioning by excitation alone. In practice, TPEM is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 limits the average input power to less than 10 mW, only slightly greater than the power normally used in confocal microscopy. Because of the intensity-squared dependence of the two-photon absorption, the excitation is limited to the focal volume.


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