scholarly journals Imaging Mitochondrial Calcium Fluxes with Fluorescent Probes and Single- or Two-Photon Confocal Microscopy

2018 ◽  
pp. 171-186 ◽  
Author(s):  
Sean M. Davidson ◽  
Michael R. Duchen
Keyword(s):  
Confocal Microscopy ◽  
Fluorescent Probes ◽  
Mitochondrial Calcium ◽  
Calcium Fluxes ◽  
Two Photon

Two-photon excitation microscopy in cellular biophysics

1995 ◽  
Vol 53 ◽  
pp. 62-63
Author(s):  
David W. Piston ◽  
Brian D. Bennett ◽  
Robert G. Summers
Keyword(s):  
Confocal Microscopy ◽  
Laser Beam ◽  
Duty Cycle ◽  
Excited Electronic State ◽  
Input Power ◽  
Two Photon ◽  
Simultaneous Absorption ◽  
Photon Excitation ◽  
Very High ◽  
Two Photon Excitation

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.

Two-photon excitation fluorescence microscopy in living systems

1993 ◽  
Vol 51 ◽  
pp. 154-155 ◽  
Author(s):  
David W. Piston
Keyword(s):  
Confocal Microscopy ◽  
Fluorescence Microscopy ◽  
Singlet State ◽  
Excited Electronic State ◽  
Input Power ◽  
Two Photon ◽  
Simultaneous Absorption ◽  
Photon Excitation ◽  
Very High ◽  
Two Photon Excitation

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.

Two-Photon Excitation Imaging of Glucose Metabolism in Living Tissue

1997 ◽  
Vol 3 (S2) ◽  
pp. 305-306
Author(s):  
David W. Piston
Keyword(s):  
Confocal Microscopy ◽  
Collection Efficiency ◽  
Three Dimensional ◽  
Spatial Filter ◽  
Input Power ◽  
Photon Absorption ◽  
Focal Volume ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

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.

Two-Photon Fluorescent Probes for Detecting Enzyme Activities in Live Tissues

2021 ◽  
Author(s):  
Vinayak Juvekar ◽  
Hyo Won Lee ◽  
Hwan Myung Kim
Keyword(s):  
Fluorescent Probes ◽  
Enzyme Activities ◽  
Two Photon

scholarly journals Polarity-active NIR probes with strong two-photon absorption and ultrahigh binding affinity of insulin amyloid fibrils

2021 ◽  
Author(s):  
Li Li ◽  
Zheng Lv ◽  
Zhongwei Man ◽  
Zhenzhen Xu ◽  
YuLing Wei ◽  
...  
Keyword(s):  
Neurodegenerative Diseases ◽  
Binding Affinity ◽  
Fluorescent Probes ◽  
Amyloid Fibrils ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Medical Diagnostic

Amyloid fibrils are associated with many neurodegenerative diseases. In-situ and in-vivo visualization of amyloid fibrils is important for medical diagnostic and requires fluorescent probes with both excitation and emission wavelengths in...

scholarly journals Real‐time two‐photon confocal microscopy using a femtosecond, amplified Ti:sapphire system

1996 ◽  
Vol 181 (3) ◽  
pp. 253-259 ◽  
Author(s):  
G. J. BRAKENHOFF ◽  
J. SQUIER ◽  
T. NORRIS ◽  
A. C. BLITON ◽  
M. H. WADE ◽  
...  
Keyword(s):  
Confocal Microscopy ◽  
Real Time ◽  
Two Photon

Imaging theory and resolution improvement of two-photon confocal microscopy

2002 ◽  
Vol 45 (11) ◽  
pp. 1468-1478 ◽  
Author(s):  
Tang Zhilie ◽  
Yang Chuping ◽  
Pei Hongjin ◽  
Liang Ruisheng ◽  
Liu Songhao
Keyword(s):  
Confocal Microscopy ◽  
Two Photon ◽  
Resolution Improvement ◽  
Imaging Theory
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