Doppler-free two-photon modulation transfer spectroscopy in sodium dimers

1990 ◽  
Vol 51 (3) ◽  
pp. 233-237 ◽  
Author(s):  
L. S. Ma ◽  
L. E. Ding ◽  
Z. Y. Bi
Keyword(s):  
Modulation Transfer ◽  
Two Photon ◽  
Sodium Dimers

Determination of the Best Emulsion for the Microscopy of Crystals

1981 ◽  
Vol 39 ◽  
pp. 32-33
Author(s):  
David A. Grano ◽  
Kenneth H. Downing
Keyword(s):  
Spatial Frequency ◽  
Information Transfer ◽  
Signal To Noise Ratio ◽  
Experimental Situation ◽  
Photographic Emulsion ◽  
Modulation Transfer ◽  
Signal To Noise ◽  
Frequency Space ◽  
Noise Ratio

The retrieval of high-resolution information from images of biological crystals depends, in part, on the use of the correct photographic emulsion. We have been investigating the information transfer properties of twelve emulsions with a view toward 1) characterizing the emulsions by a few, measurable quantities, and 2) identifying the “best” emulsion of those we have studied for use in any given experimental situation. Because our interests lie in the examination of crystalline specimens, we've chosen to evaluate an emulsion's signal-to-noise ratio (SNR) as a function of spatial frequency and use this as our critereon for determining the best emulsion.The signal-to-noise ratio in frequency space depends on several factors. First, the signal depends on the speed of the emulsion and its modulation transfer function (MTF). By procedures outlined in, MTF's have been found for all the emulsions tested and can be fit by an analytic expression 1/(1+(S/S0)2). Figure 1 shows the experimental data and fitted curve for an emulsion with a better than average MTF. A single parameter, the spatial frequency at which the transfer falls to 50% (S0), characterizes this curve.

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.

P20 phosphor on polyimide as a large area high-resolution transmission screen for slow scan CCD systems

1995 ◽  
Vol 53 ◽  
pp. 20-21
Author(s):  
C. C. Ahn ◽  
S. Karnes ◽  
M. Lvovsky ◽  
C. M. Garland ◽  
H. A. Atwater ◽  
...  
Keyword(s):  
Mechanical Stability ◽  
High Energy ◽  
Practical Implementation ◽  
Line Pair ◽  
Modulation Transfer ◽  
Large Area ◽  
Ccd Imaging ◽  
Transmission Electron ◽  
The One ◽  
Beam Broadening

The bane of CCD imaging systems for transmission electron microscopy at intermediate and high voltages has been their relatively poor modulation transfer function (MTF), or line pair resolution. The problem originates primarily with the phosphor screen. On the one hand, screens should be thick so that as many incident electrons as possible are converted to photons, yielding a high detective quantum efficiency(DQE). The MTF diminishes as a function of scintillator thickness however, and to some extent as a function of fluorescence within the scintillator substrates. Fan has noted that the use of a thin layer of phosphor beneath a self supporting 2μ, thick Al substrate might provide the most appropriate compromise for high DQE and MTF in transmission electron microcscopes which operate at higher voltages. Monte Carlo simulations of high energy electron trajectories reveal that only little beam broadening occurs within this thickness of Al film. Consequently, the MTF is limited predominantly by broadening within the thin phosphor underlayer. There are difficulties however, in the practical implementation of this design, associated mostly with the mechanical stability of the Al support film.

Spot-scan imaging of ice-embedded catalase crystal on a slow-scan CCD camera in a 400-kV electron cryomicroscope

1994 ◽  
Vol 52 ◽  
pp. 98-99
Author(s):  
Wah Chiu ◽  
Michael Sherman ◽  
Jaap Brink
Keyword(s):  
Electron Diffraction ◽  
Ccd Camera ◽  
Protein Crystal ◽  
Modulation Transfer ◽  
Measured Intensity ◽  
3 Dimensional ◽  
Dimensional Reconstruction ◽  
Diffraction Patterns ◽  
Complex Crystal

In protein electron crystallography, both low dose electron diffraction patterns and images are needed to provide accurate amplitudes and phases respectively for a 3-dimensional reconstruction. We have demonstrated that the Gatan 1024x1024 model 679 slow-scan CCD camera is useful to record electron diffraction intensities of glucose-embedded crotoxin complex crystal to 3 Å resolution. The quality of the electron diffraction intensities is high on the basis of the measured intensity equivalence ofthe Friedel-related reflections. Moreover, the number of patterns recorded from a single crystal can be as high as 120 under the constraints of radiation damage and electron statistics for the reflections in each pattern.A limitation of the slow-scan CCD camera for recording electron images of protein crystal arises from the relatively large pixel size, i.e. 24 μm (provided by Gatan). The modulation transfer function of our camera with a P43 scintillator has been determined for 400 keV electrons and shows an amplitude fall-off to 0.25 at 1/60 μm−1.

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.

Analysis of photographic emulsions for High-Voltage Electron Microscopy

1993 ◽  
Vol 51 ◽  
pp. 452-453 ◽  
Author(s):  
James R. Kremer ◽  
Paul S. Furcinitti ◽  
Eileen O’Toole ◽  
J. Richard McIntosh
Keyword(s):  
Electron Microscope ◽  
Optical Density ◽  
High Voltage ◽  
Noise Power ◽  
Limited Information ◽  
Modulation Transfer ◽  
Transmission Microscopy ◽  
High Voltage Electron Microscopy ◽  
Voltage Range ◽  
Voltage Electron

Characteristics of electron microscope film emulsions, such as the speed, the modulation transfer function, and the exposure dependence of the noise power spectrum, have been studied for electron energies (80-100keV) used in conventional transmission microscopy. However, limited information is available for electron energies in the intermediate to high voltage range, 300-1000keV. Furthermore, emulsion characteristics, such as optical density versus exposure, for new or improved emulsions are usually only quoted by film manufacturers for 80keV electrons. The need for further film emulsion studies at higher voltages becomes apparent when searching for a film to record low dose images of radiation sensitive biological specimens in the frozen hydrated state. Here, we report the optical density, speed and relative resolution of a few of the more popular electron microscope films after exposure to 1MeV electrons.Three electron microscope films, Kodak S0-163, Kodak 4489, and Agfa Scientia 23D56 were tested with a JEOLJEM-1000 electron microscope operating at an accelerating voltage of 1000keV.

Second order interference in two photon absorption

1996 ◽  
Vol 43 (9) ◽  
pp. 1765-1771 ◽  
Author(s):  
M. W. HAMILTON and D. S. ELLIOTT
Keyword(s):  
Second Order ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon
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