Cryophotolysis of a caged oxygen compound for use in low temperature biological studies

2009 ◽  
Vol 8 (8) ◽  
pp. 1150 ◽  
Author(s):  
Annaleise R. Howard-Jones ◽  
Virgile Adam ◽  
Andrew Cowley ◽  
Jack E. Baldwin ◽  
Dominique Bourgeois
Keyword(s):  
Low Temperature ◽  
Oxygen Compound ◽  
Biological Studies

scholarly journals The reaction of cytochrome o in Escherichia coli with oxygen. Low-temperature kinetic and spectral studies

1979 ◽  
Vol 184 (2) ◽  
pp. 379-389 ◽  
Author(s):  
R K Poole ◽  
A J Waring ◽  
B Chance
Keyword(s):  
Escherichia Coli ◽  
Low Temperature ◽  
Flash Photolysis ◽  
Spectral Characteristics ◽  
Spectral Studies ◽  
Oxygen Compound ◽  
Intact Cells ◽  
Lower Velocity ◽  
Cytochrome D ◽  
O2 Binding

1. The reactions of cytochrome omicron in intact cells of aerobically grown Escherichia coli with O2 and CO have been studied at low temperature. 2. Flash photolysis of CO-liganded cells in the presence of O2 and at temperatures between −79 and −102 degrees C results in the oxidation of kinetically heterogeneous beta-type cytochromes (including cytochrome omicron), but not of cytochrome d. 3. The reaction of reduced cytochrome omicron with O2 involves O2 binding to give intermediate(s) with spectral characteristics similar to that of the reduced oxidase-CO complex. Observation in the alpha-region suggests that unexplained ligand dissociation accompanies the initial O2 binding. 4. At temperatures below −98 degrees C, an ‘end point’ in the reaction is reached; further reaction and oxidation of cytochrome omicron occurs on raising the temperature. 5. There is a linear relationship between the rate of formation of the oxygen compound and the O2 concentration up to 0.5 mM. The second-order constant for its formation (k+1) is 0.91 M−1.S−1 at −101 degrees C. The reaction is not readily reversible, the value of k−1 being 1.4 × 10(-5) S−1 and the kd 1.5 × 10(-5) M. 6. The energy of activation for this reaction at low temperatures is 29.9kJ (7.1 kcal)/mol. 7. The reaction with O2 is distinguished from that with CO by the markedly lower velocity and high photolytic reversibility of the latter. 8. Comparisons are drawn between the intermediate(s) in the O2 reaction of cytochrome omicron in E. coli and those identified in other bacteria and in the reaction of cytochrome aa3 with O2.

Synthesis, Spectral, Electrochemical, Magnetic and Biological Studies of Tet-A Based Binuclear Mn(III) Complexes

2012 ◽  
Vol 584 ◽  
pp. 386-390
Author(s):  
R. Prabu ◽  
L. Vijayalakshmi ◽  
V. Narayanan
Keyword(s):  
Spectral Analysis ◽  
Schiff Base ◽  
Low Temperature ◽  
Condensation Reaction ◽  
Magnetic Behavior ◽  
Redox Processes ◽  
Antiferromagnetic Exchange ◽  
Electrochemical Studies ◽  
Biological Studies ◽  
Perchlorate Hexahydrate

A new phenoxo-bridged Tet-a based binuclear manganese(III) complexes were synthesized by the Schiff base condensation reaction of the compound 1,8–[bis(3-formyl-2-hydroxy-5-methyl)benzyl] -5,5,7,12,12,14- hexamethyl -l,4,8,11-tetraazacyclotetradecane (PC–a) and 1,8– [bis(3-formyl-2-hydroxy-5-bromo)benzyl]- 5,5,7,12,12,14-hexamethyl-l,4,8,11-tetraazacyclotetradecane (PC–b) with triethylenetetramine and manganese(II) perchlorate hexahydrate. Their synthesized complexes were confirmed by analytical and spectral analysis. The phenolate oxygen atom is acting as bridge between two unsymmetrically coordinated Mn atoms. Electrochemical studies of the complexes reveal that all the redox processes are irreversible in nature. Low temperature magnetic study shows that the complexes exhibit antiferromagnetic exchange interaction between the two metal ions. The spectral and magnetic behavior of the complexes is discussed.

Modification of the Electron Microscope for Investigation of Fully Hydrated Biological Specimens

1969 ◽  
Vol 27 ◽  
pp. 418-419 ◽  
Author(s):  
R. C. Moretz ◽  
D. F. Parsons
Keyword(s):  
Electron Beam ◽  
Electron Microscope ◽  
Structural Damage ◽  
Lower Percentage ◽  
Vacuum Drying ◽  
Total Removal ◽  
Total Absence ◽  
Biological Studies ◽  
Electron Microscopes ◽  
Beam Damage

Short lifetime or total absence of electron diffraction of ordered biological specimens is an indication that the specimen undergoes extensive molecular structural damage in the electron microscope. The specimen damage is due to the interaction of the electron beam (40-100 kV) with the specimen and the total removal of water from the structure by vacuum drying. The lower percentage of inelastic scattering at 1 MeV makes it possible to minimize the beam damage to the specimen. The elimination of vacuum drying by modification of the electron microscope is expected to allow more meaningful investigations of biological specimens at 100 kV until 1 MeV electron microscopes become more readily available. One modification, two-film microchambers, has been explored for both biological and non-biological studies.

Exsolution and inversion in pigeonite from the Whin Sill, Northern England

1978 ◽  
Vol 36 (1) ◽  
pp. 474-475
Author(s):  
P.P.K. Smith
Keyword(s):  
High Temperature ◽  
Low Temperature ◽  
Homogeneous Nucleation ◽  
Silicate Mineral ◽  
Antiphase Boundaries ◽  
Two Phase ◽  
Primitive Form ◽  
The Core ◽  
Nucleation Mechanisms ◽  
And Inversion

Grains of pigeonite, a calcium-poor silicate mineral of the pyroxene group, from the Whin Sill dolerite have been ion-thinned and examined by TEM. The pigeonite is strongly zoned chemically from the composition Wo8En64FS28 in the core to Wo13En34FS53 at the rim. Two phase transformations have occurred during the cooling of this pigeonite:- exsolution of augite, a more calcic pyroxene, and inversion of the pigeonite from the high- temperature C face-centred form to the low-temperature primitive form, with the formation of antiphase boundaries (APB's). Different sequences of these exsolution and inversion reactions, together with different nucleation mechanisms of the augite, have created three distinct microstructures depending on the position in the grain.In the core of the grains small platelets of augite about 0.02μm thick have farmed parallel to the (001) plane (Fig. 1). These are thought to have exsolved by homogeneous nucleation. Subsequently the inversion of the pigeonite has led to the creation of APB's.

Low temperature x-ray microanalysis of frozen-hydrated tobacco leaves

1984 ◽  
Vol 42 ◽  
pp. 284-285
Author(s):  
S. Edith Taylor ◽  
Patrick Echlin ◽  
May McKoon ◽  
Thomas L. Hayes
Keyword(s):  
Low Temperature ◽  
Lemna Minor ◽  
Root Tips ◽  
Tobacco Leaves ◽  
X Ray ◽  
Whole Cells ◽  
Carbon Coated ◽  
The Right ◽  
Beam Currents ◽  
The University

Low temperature x-ray microanalysis (LTXM) of solid biological materials has been documented for Lemna minor L. root tips. This discussion will be limited to a demonstration of LTXM for measuring relative elemental distributions of P,S,Cl and K species within whole cells of tobacco leaves.Mature Wisconsin-38 tobacco was grown in the greenhouse at the University of California, Berkeley and picked daily from the mid-stalk position (leaf #9). The tissue was excised from the right of the mid rib and rapidly frozen in liquid nitrogen slush. It was then placed into an Amray biochamber and maintained at 103K. Fracture faces of the tissue were prepared and carbon-coated in the biochamber. The prepared sample was transferred from the biochamber to the Amray 1000A SEM equipped with a cold stage to maintain low temperatures at 103K. Analyses were performed using a tungsten source with accelerating voltages of 17.5 to 20 KV and beam currents from 1-2nA.

Bulk standards for low-temperature biological x-ray microanalysis

1984 ◽  
Vol 42 ◽  
pp. 282-283
Author(s):  
P. Echlin ◽  
M. McKoon ◽  
E.S. Taylor ◽  
C.E. Thomas ◽  
K.L. Maloney ◽  
...  
Keyword(s):  
Phase Separation ◽  
Low Temperature ◽  
Analytical Method ◽  
Salt Solutions ◽  
Absolute Concentration ◽  
Cooling Process ◽  
X Ray ◽  
The Absolute ◽  
Analytical Procedures ◽  
Electrical Conductors

Although sections of frozen salt solutions have been used as standards for x-ray microanalysis, such solutions are less useful when analysed in the bulk form. They are poor thermal and electrical conductors and severe phase separation occurs during the cooling process. Following a suggestion by Whitecross et al we have made up a series of salt solutions containing a small amount of graphite to improve the sample conductivity. In addition, we have incorporated a polymer to ensure the formation of microcrystalline ice and a consequent homogenity of salt dispersion within the frozen matrix. The mixtures have been used to standardize the analytical procedures applied to frozen hydrated bulk specimens based on the peak/background analytical method and to measure the absolute concentration of elements in developing roots.

Field ion microscopy

1988 ◽  
Vol 46 ◽  
pp. 434-435
Author(s):  
Gert Ehrlich
Keyword(s):  
Low Temperature ◽  
Sample Surface ◽  
Low Pressure ◽  
Radius Of Curvature ◽  
Field Ion Microscopy ◽  
Phosphor Screen ◽  
Ion Microscopy ◽  
Field Ion Microscope

The field ion microscope, devised by Erwin Muller in the 1950's, was the first instrument to depict the structure of surfaces in atomic detail. An FIM image of a (111) plane of tungsten (Fig.l) is typical of what can be done by this microscope: for this small plane, every atom, at a separation of 4.48Å from its neighbors in the plane, is revealed. The image of the plane is highly enlarged, as it is projected on a phosphor screen with a radius of curvature more than a million times that of the sample. Müller achieved the resolution necessary to reveal individual atoms by imaging with ions, accommodated to the object at a low temperature. The ions are created at the sample surface by ionization of an inert image gas (usually helium), present at a low pressure (< 1 mTorr). at fields on the order of 4V/Å.

High-Vacuum Evaporation and a Cryostage to Observe Fractured Yeast

1988 ◽  
Vol 46 ◽  
pp. 256-257
Author(s):  
William P. Wergin ◽  
Eric F. Erbe ◽  
Eugene L. Vigil
Keyword(s):  
Electron Microscope ◽  
Low Temperature ◽  
Standard Specimen ◽  
High Vacuum ◽  
Specimen Holder ◽  
Sputter Coating ◽  
Fresh Frozen ◽  
Gain Access ◽  
Scanning Electron ◽  
Transfer Device

Investigators have long realized the potential advantages of using a low temperature (LT) stage to examine fresh, frozen specimens in a scanning electron microscope (SEM). However, long working distances (W.D.), thick sputter coatings and surface contamination have prevented LTSEM from achieving results comparable to those from TEM freeze etch. To improve results, we recently modified techniques that involve a Hitachi S570 SEM, an Emscope SP2000 Sputter Cryo System and a Denton freeze etch unit. Because investigators have frequently utilized the fractured E face of the plasmalemma of yeast, this tissue was selected as a standard for comparison in the present study.In place of a standard specimen holder, a modified rivet was used to achieve a shorter W.D. (1 to -2 mm) and to gain access to the upper detector. However, the additional height afforded by the rivet, precluded use of the standard shroud on the Emscope specimen transfer device. Consequently, the sample became heavily contaminated (Fig. 1). A removable shroud was devised and used to reduce contamination (Fig. 2), but the specimen lacked clean fractured edges. This result suggested that low vacuum sputter coating was also limiting resolution.

Observation by HVEM of the martensite transformation and the superconducting transition in Nb3Sn

1988 ◽  
Vol 46 ◽  
pp. 788-789
Author(s):  
M. A. Kirk ◽  
M. C. Baker ◽  
B. J. Kestel ◽  
H. W. Weber
Keyword(s):  
Thin Films ◽  
Low Temperature ◽  
Superconducting State ◽  
Martensite Transformation ◽  
Sputter Deposition ◽  
Diffusion Reaction ◽  
Solute Diffusion ◽  
Superconducting Transition ◽  
Twin Boundaries ◽  
Martensite Structure

It is well known that a number of compound superconductors with the A15 structure undergo a martensite transformation when cooled to the superconducting state. Nb3Sn is one of those compounds that transforms, at least partially, from a cubic to tetragonal structure near 43 K. To our knowledge this transformation in Nb3Sn has not been studied by TEM. In fact, the only low temperature TEM study of an A15 material, V3Si, was performed by Goringe and Valdre over 20 years ago. They found the martensite structure in some foil areas at temperatures between 11 and 29 K, accompanied by faults that consisted of coherent twin boundaries on {110} planes. In pursuing our studies of irradiation defects in superconductors, we are the first to observe by TEM a similar martensite structure in Nb3Sn.Samples of Nb3Sn suitable for TEM studies have been produced by both a liquid solute diffusion reaction and by sputter deposition of thin films.

Design of Sensitive Photographic Materials for the High-Voltage Electron Microscope

1975 ◽  
Vol 33 ◽  
pp. 156-157
Author(s):  
Murray Vernon King ◽  
Donald F. Parsons
Keyword(s):  
Electron Microscope ◽  
Radiation Damage ◽  
High Voltage ◽  
Radiation Sensitivity ◽  
Fast Electrons ◽  
Voltage Electron ◽  
High Voltage Electron Microscope ◽  
High Voltage Electron ◽  
Biological Studies ◽  
Low Sensitivity

Effective application of the high-voltage electron microscope to a wide variety of biological studies has been restricted by the radiation sensitivity of biological systems. The problem of radiation damage has been recognized as a serious factor influencing the amount of information attainable from biological specimens in electron microscopy at conventional voltages around 100 kV. The problem proves to be even more severe at higher voltages around 1 MV. In this range, the problem is the relatively low sensitivity of the existing recording media, which entails inordinately long exposures that give rise to severe radiation damage. This low sensitivity arises from the small linear energy transfer for fast electrons. Few developable grains are created in the emulsion per electron, while most of the energy of the electrons is wasted in the film base.

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