The Generation and Characterization of Ultrashort Electron Pulses and their Application to Time-Resolved Electron Diffraction.

1996 ◽  
Author(s):  
Peter M. Weber
Keyword(s):  
Electron Diffraction ◽  
Time Resolved ◽  
Electron Pulses

scholarly journals Structural and Spectroscopic Characterization of Photoactive Yellow Protein and Photoswitchable Fluorescent Protein Constructs Containing Heavy Atoms

2020 ◽  
Author(s):  
Matthew Romei ◽  
Chi-Yun Lin ◽  
Steven Boxer
Keyword(s):  
Electron Diffraction ◽  
Fluorescent Protein ◽  
Heavy Atom ◽  
Spectroscopic Studies ◽  
Photoactive Yellow Protein ◽  
Structural Rearrangements ◽  
Time Resolved ◽  
Heavy Atoms ◽  
Protein Constructs

Photo-induced structural rearrangements of chromophore-containing proteins are essential for various light-dependent signaling pathways and optogenetic applications. Ultrafast structural and spectroscopic methods have offered insights into these structural rearrangements across many timescales. However, questions still remain about exact mechanistic details, especially regarding photoisomerization of the chromophore within these proteins femtoseconds to picoseconds after photoexcitation. Instrumentation advancements for time-resolved crystallography and ultrafast electron diffraction provide a promising opportunity to study these reactions, but achieving enough signal-to-noise is a constant challenge. Here we present four new photoactive yellow protein constructs and one new fluorescent protein construct that contain heavy atoms either within or around the chromophore and can be expressed with high yields. Structural characterization of these constructs, most at atomic resolution, show minimal perturbation caused by the heavy atoms compared to wild-type structures. Spectroscopic studies report the effects of the heavy atom identity and location on the chromophore’s photophysical properties. None of the substitutions prevent photoisomerization, although certain rates within the photocycle may be affected. Overall, these new proteins containing heavy atoms are ideal samples for state-of-the-art time-resolved crystallography and electron diffraction experiments to elucidate crucial mechanistic information of photoisomerization.

scholarly journals Structural and Spectroscopic Characterization of Photoactive Yellow Protein and Photoswitchable Fluorescent Protein Constructs Containing Heavy Atoms

2020 ◽  
Author(s):  
Matthew Romei ◽  
Chi-Yun Lin ◽  
Steven Boxer
Keyword(s):  
Electron Diffraction ◽  
Fluorescent Protein ◽  
Heavy Atom ◽  
Spectroscopic Studies ◽  
Photoactive Yellow Protein ◽  
Structural Rearrangements ◽  
Time Resolved ◽  
Heavy Atoms ◽  
Protein Constructs

Photo-induced structural rearrangements of chromophore-containing proteins are essential for various light-dependent signaling pathways and optogenetic applications. Ultrafast structural and spectroscopic methods have offered insights into these structural rearrangements across many timescales. However, questions still remain about exact mechanistic details, especially regarding photoisomerization of the chromophore within these proteins femtoseconds to picoseconds after photoexcitation. Instrumentation advancements for time-resolved crystallography and ultrafast electron diffraction provide a promising opportunity to study these reactions, but achieving enough signal-to-noise is a constant challenge. Here we present four new photoactive yellow protein constructs and one new fluorescent protein construct that contain heavy atoms either within or around the chromophore and can be expressed with high yields. Structural characterization of these constructs, most at atomic resolution, show minimal perturbation caused by the heavy atoms compared to wild-type structures. Spectroscopic studies report the effects of the heavy atom identity and location on the chromophore’s photophysical properties. None of the substitutions prevent photoisomerization, although certain rates within the photocycle may be affected. Overall, these new proteins containing heavy atoms are ideal samples for state-of-the-art time-resolved crystallography and electron diffraction experiments to elucidate crucial mechanistic information of photoisomerization.

scholarly journals A Compact Ultrafast Electron Diffractometer with Relativistic Femtosecond Electron Pulses

2020 ◽  
Vol 4 (1) ◽  
pp. 4 ◽  
Author(s):  
Jinfeng Yang ◽  
Kazuki Gen ◽  
Nobuyasu Naruse ◽  
Shouichi Sakakihara ◽  
Yoichi Yoshida
Keyword(s):  
Electron Diffraction ◽  
Amorphous Materials ◽  
Crystalline Silicon ◽  
Electron Gun ◽  
Diffraction Measurement ◽  
Single Shot ◽  
Time Resolved ◽  
Single Crystalline ◽  
Electron Lens ◽  
Electron Pulses

We have developed a compact relativistic femtosecond electron diffractometer with a radio-frequency photocathode electron gun and an electron lens system. The electron gun generated 2.5-MeV-energy electron pulses with a duration of 55 ± 5 fs containing 6.3 × 104 electrons per pulse. Using these pulses, we successfully detected high-contrast electron diffraction images of single crystalline, polycrystalline, and amorphous materials. An excellent spatial resolution of diffraction images was obtained as 0.027 ± 0.001 Å−1. In the time-resolved electron diffraction measurement, a laser-excited ultrafast electronically driven phase transition in single-crystalline silicon was observed with a temporal resolution of 100 fs. The results demonstrate the advantages of the compact relativistic femtosecond electron diffractometer, including access to high-order Bragg reflections, single shot imaging with the relativistic femtosecond electron pulse, and the feasibility of time-resolved electron diffraction to study ultrafast structural dynamics.

Electron Channeling Patterns

1977 ◽  
Vol 35 ◽  
pp. 12-13
Author(s):  
David C. Joy
Keyword(s):  
Electron Diffraction ◽  
Incidence Angle ◽  
Photographic Plate ◽  
Diffraction Effect ◽  
Angle Of Incidence ◽  
Bulk Single Crystal ◽  
Time Resolved ◽  
Electron Channeling ◽  
Spatially Resolved ◽  
Large Bulk

Electron channeling patterns (ECP) were first found by Coates (1967) while observing a large bulk, single crystal of silicon in a scanning electron microscope. The geometric pattern visible was shown to be produced as a result of the changes in the angle of incidence, between the beam and the specimen surface normal, which occur when the sample is examined at low magnification (Booker, Shaw, Whelan and Hirsch 1967).A conventional electron diffraction pattern consists of an angularly resolved intensity distribution in space which may be directly viewed on a fluorescent screen or recorded on a photographic plate. An ECP, on the other hand, is produced as the result of changes in the signal collected by a suitable electron detector as the incidence angle is varied. If an integrating detector is used, or if the beam traverses the surface at a fixed angle, then no channeling contrast will be observed. The ECP is thus a time resolved electron diffraction effect. It can therefore be related to spatially resolved diffraction phenomena by an application of the concepts of reciprocity (Cowley 1969).

Characterization of submicrometer fluid Inclusions in minerals by Analytical/Transmission Electron Microscopy and electron diffraction

1990 ◽  
Vol 48 (4) ◽  
pp. 424-424
Author(s):  
George Guthrie ◽  
David Veblen
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Electron Microscope ◽  
Electron Diffraction ◽  
Light Microscopy ◽  
Fluid Inclusions ◽  
Energy Dispersive Spectrometer ◽  
Analytical Transmission Electron Microscopy ◽  
Transmission Electron

The nature of a geologic fluid can often be inferred from fluid-filled cavities (generally <100 μm in size) that are trapped during the growth of a mineral. A variety of techniques enables the fluids and daughter crystals (any solid precipitated from the trapped fluid) to be identified from cavities greater than a few micrometers. Many minerals, however, contain fluid inclusions smaller than a micrometer. Though inclusions this small are difficult or impossible to study by conventional techniques, they are ideally suited for study by analytical/ transmission electron microscopy (A/TEM) and electron diffraction. We have used this technique to study fluid inclusions and daughter crystals in diamond and feldspar.Inclusion-rich samples of diamond and feldspar were ion-thinned to electron transparency and examined with a Philips 420T electron microscope (120 keV) equipped with an EDAX beryllium-windowed energy dispersive spectrometer. Thin edges of the sample were perforated in areas that appeared in light microscopy to be populated densely with inclusions. In a few cases, the perforations were bound polygonal sides to which crystals (structurally and compositionally different from the host mineral) were attached (Figure 1).

scholarly journals Characterization of a Novel FRET Probe in a Crowded Environment using Time-Resolved Anisotropy

2017 ◽  
Vol 112 (3) ◽  
pp. 346a
Author(s):  
Hannah Leopold ◽  
Megan Currie ◽  
Jacob Schwarz ◽  
Arnold J. Boersma ◽  
Erin D. Sheets ◽  
...  
Keyword(s):  
Time Resolved ◽  
Crowded Environment

scholarly journals Synthesis and characterization of Fe, Co, and Ni colloids in 2-mercaptoethanol

2020 ◽  
Vol 10 ◽  
pp. 184798042096688
Author(s):  
Galo Cárdenas-Triviño ◽  
Sergio Triviño-Matus
Keyword(s):  
Electron Microscopy ◽  
Electron Diffraction ◽  
Colloidal Dispersions ◽  
Synthesis And Characterization ◽  
X Ray ◽  
Transmission Electron ◽  
Vis Spectroscopy ◽  
Chemical Liquid Deposition ◽  
Liquid Deposition

Metal colloids in 2-mercaptoethanol using nanoparticles (NPs) of iron (Fe), cobalt (Co), and nickel (Ni) were prepared by chemical liquid deposition method. Transmission electron microscopy, electron diffraction, UV-VIS spectroscopy, and scanning electron microscopy with electron dispersive X-ray spectroscopy characterized the resulting colloidal dispersions. The NPs exhibited sizes with ranges from 9.8 nm for Fe, 3.7 nm for Co, and 7.2 nm for Ni. The electron diffraction shows the presence of the metals in its elemental state Fe (0), Co (0), and Ni (0) and also some compounds FeO (OH), CoCo2S4, and NiNi2S4.

scholarly journals KD determination from time-resolved experiments on live cells with LigandTracer and reconciliation with end-point flow cytometry measurements

2021 ◽  
Author(s):  
Diana Spiegelberg ◽  
Jonas Stenberg ◽  
Pascale Richalet ◽  
Marc Vanhove
Keyword(s):  
Flow Cytometry ◽  
Live Cells ◽  
Time Resolved ◽  
Surface Receptors ◽  
Full Characterization ◽  
End Point ◽  
Titration Curves ◽  
Kinetics Of

AbstractDesign of next-generation therapeutics comes with new challenges and emulates technology and methods to meet them. Characterizing the binding of either natural ligands or therapeutic proteins to cell-surface receptors, for which relevant recombinant versions may not exist, represents one of these challenges. Here we report the characterization of the interaction of five different antibody therapeutics (Trastuzumab, Rituximab, Panitumumab, Pertuzumab, and Cetuximab) with their cognate target receptors using LigandTracer. The method offers the advantage of being performed on live cells, alleviating the need for a recombinant source of the receptor. Furthermore, time-resolved measurements, in addition to allowing the determination of the affinity of the studied drug to its target, give access to the binding kinetics thereby providing a full characterization of the system. In this study, we also compared time-resolved LigandTracer data with end-point KD determination from flow cytometry experiments and hypothesize that discrepancies between these two approaches, when they exist, generally come from flow cytometry titration curves being acquired prior to full equilibration of the system. Our data, however, show that knowledge of the kinetics of the interaction allows to reconcile the data obtained by flow cytometry and LigandTracer and demonstrate the complementarity of these two methods.

scholarly journals Time-Resolved Spectroscopic Study of N,N-Di(4-bromo)nitrenium Ions in Selected Solutions

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3182 ◽  
Author(s):  
Lili Du ◽  
Xin Lan ◽  
Zhiping Yan ◽  
Ruixue Zhu ◽  
David Phillips
Keyword(s):  
Transient Absorption ◽  
Direct Detection ◽  
Reaction Pathway ◽  
Addition Reactions ◽  
Time Resolved ◽  
Nitrenium Ions ◽  
Future Studies ◽  
Nitrenium Ion ◽  
Design Time

Nitrenium ions are important reactive intermediates in chemistry and biology. In this work, femtosecond and nanosecond transient absorption (fs-TA and ns-TA) along with nanosecond time-resolved resonance Raman (ns-TR3) experiments were employed to examine the photochemical pathways of N-(4,4′-dibromodiphenylamino)-2,4,6-trimethylpyridinium BF4− (salt (DN) from just absorption of a photon of light to the production of the important N,N-di(4-bromophenyl)nitrenium ion 2. In acetonitrile (MeCN), the formation of halogenated diarylnitrenium ion 2 was observed within 4 ps, showing the vibrational spectra with strong intensity. The nucleophilic adduct reaction of ion 2 with H2O was also examined in aqueous solutions. The direct detection of the unique ortho adduct intermediate 3 shows that there is an efficient and exclusive reaction pathway for 2 with H2O. The results shown in this paper give new characterization of 2, which can be used to design time-resolved spectroscopy investigations of covalent addition reactions of nitrenium ions with other molecules in future studies.

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