Two‐photon excitation 4Pi confocal microscope: Enhanced axial resolution microscope for biological research

1995 ◽  
Vol 66 (13) ◽  
pp. 1698-1700 ◽  
Author(s):  
P. E. Hänninen ◽  
S. W. Hell ◽  
J. Salo ◽  
E. Soini ◽  
C. Cremer
Keyword(s):  
Confocal Microscope ◽  
Biological Research ◽  
Axial Resolution ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

scholarly journals Axial Resolution Enhancement by 4Pi Confocal Fluorescence Microscopy with Two-Photon Excitation

2007 ◽  
Vol 33 (5-6) ◽  
pp. 433-443 ◽  
Author(s):  
Sylvia Glaschick ◽  
Carlheinz Röcker ◽  
Karen Deuschle ◽  
Jörg Wiedenmann ◽  
Franz Oswald ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Resolution Enhancement ◽  
Confocal Fluorescence Microscopy ◽  
Axial Resolution ◽  
Two Photon ◽  
Photon Excitation ◽  
Confocal Fluorescence ◽  
Two Photon Excitation

Millisecond lifetime imaging with a europium complex using a commercial confocal microscope under one or two-photon excitation

2014 ◽  
Vol 5 (9) ◽  
pp. 3475-3485 ◽  
Author(s):  
Alexei Grichine ◽  
Alexandre Haefele ◽  
Simon Pascal ◽  
Alain Duperray ◽  
Richard Michel ◽  
...  
Keyword(s):  
Confocal Microscope ◽  
Europium Complex ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Long Lifetime ◽  
Two Photon Excitation

We demonstrate that a commercial confocal microscope can perform time-gated and long lifetime imaging in the μs to ms range under classical one photon or nonlinear two photon excitation.

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 fluorescence microscopy using a semiconductor laser

Bioimaging ◽  
1995 ◽  
Vol 3 (2) ◽  
pp. 70-75 ◽  
Author(s):  
Pekka E Hänninen ◽  
Martin Schrader ◽  
Erkki Soini ◽  
Stefan W Hell
Keyword(s):  
Fluorescence Microscopy ◽  
Semiconductor Laser ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

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.

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