Hasil untuk "Microscopy"

M. Haider, S. Uhlemann, E. Schwan et al.

F. Giessibl

N. Kodera, D. Yamamoto, R. Ishikawa et al.

Myung K. Kim

D. Axelrod

P. Midgley, M. Weyland

E. Rittweger, K. Y. Han, S. E. Irvine et al.

G. Gordon, G. Berry, X. Liang et al.

Fluorescence resonance energy transfer (FRET) is a technique used for quantifying the distance between two molecules conjugated to different fluorophores. By combining optical microscopy with FRET it is possible to obtain quantitative temporal and spatial information about the binding and interaction of proteins, lipids, enzymes, DNA, and RNA in vivo. In conjunction with the recent development of a variety of mutant green fluorescent proteins (mtGFPs), FRET microscopy provides the potential to measure the interaction of intracellular molecular species in intact living cells where the donor and acceptor fluorophores are actually part of the molecules themselves. However, steady-state FRET microscopy measurements can suffer from several sources of distortion, which need to be corrected. These include direct excitation of the acceptor at the donor excitation wavelengths and the dependence of FRET on the concentration of acceptor. We present a simple method for the analysis of FRET data obtained with standard filter sets in a fluorescence microscope. This method is corrected for cross talk (any detection of donor fluorescence with the acceptor emission filter and any detection of acceptor fluorescence with the donor emission filter), and for the dependence of FRET on the concentrations of the donor and acceptor. Measurements of the interaction of the proteins Bcl-2 and Beclin (a recently identified Bcl-2 interacting protein located on chromosome 17q21), are shown to document the accuracy of this approach for correction of donor and acceptor concentrations, and cross talk between the different filter units.

M. Juette, T. Gould, M. Lessard et al.

A. Zewail

M. Levoy, Ren Ng, Andrew Adams et al.

W. Chao, W. Chao, B. Harteneck et al.

K. Steingart, M. Henry, Vivienne Ng et al.

P. Kner, Bryant B. Chhun, E. Griffis et al.

Structured-illumination microscopy can double the resolution of the widefield fluorescence microscope but has previously been too slow for dynamic live imaging. Here we demonstrate a high-speed structured-illumination microscope that is capable of 100-nm resolution at frame rates up to 11 Hz for several hundred time points. We demonstrate the microscope by video imaging of tubulin and kinesin dynamics in living Drosophila melanogaster S2 cells in the total internal reflection mode.

Zhuo Wang, L. Millet, Mustafa Mir et al.

Spatial light interference microscopy (SLIM) is a new optical microscopy technique, capable of measuring nanoscale structures and dynamics in live cells via interferometry. SLIM combines two classic ideas in light imaging: Zernike's phase contrast microscopy, which renders high contrast intensity images of transparent specimens, and Gabor's holography, where the phase information from the object is recorded. Thus, SLIM reveals the intrinsic contrast of cell structures and, in addition, renders quantitative optical path-length maps across the sample. The resulting topographic accuracy is comparable to that of atomic force microscopy, while the acquisition speed is 1,000 times higher. We illustrate the novel insight into cell dynamics via SLIM by experiments on primary cell cultures from the rat brain. SLIM is implemented as an add-on module to an existing phase contrast microscope, which may prove instrumental in impacting the light microscopy field at a large scale.

Ricardo Garcia, E. T. Herruzo

C. Freudiger, Sijia Lu, Wei Min et al.

R. Henderson, A. Sali, M. Baker et al.

This Meeting Review describes the proceedings and conclusions from the inaugural meeting of the Electron Microscopy Validation Task Force organized by the Unified Data Resource for 3DEM (http://www.emdatabank.org) and held at Rutgers University in New Brunswick, NJ on September 28 and 29, 2010. At the workshop, a group of scientists involved in collecting electron microscopy data, using the data to determine three-dimensional electron microscopy (3DEM) density maps, and building molecular models into the maps explored how to assess maps, models, and other data that are deposited into the Electron Microscopy Data Bank and Protein Data Bank public data archives. The specific recommendations resulting from the workshop aim to increase the impact of 3DEM in biology and medicine.

Chris Xu, F. Wise

Xiao Chen, Chengdi Li, Shuangxia Zhu et al.

This study employs numerical simulations and experiments to examine the cold spray deposition of nanostructured hydroxyapatite (Ca₁₀(PO4)₆(OH)₂, HA)/70wt.%Ti composite particles under different processing conditions, based on the features of nanocomposites that strengthen interfacial adhesion and improve coating interfacial strength. Using ABAQUS/CAE combined with LS-PrePost 4.9-x64 software, the deposition behavior of the composite particles during deposition under various impact velocities was analyzed, along with the stress of the HA and Ti particles within the composite particle. The deposition behavior of both single and multiple composite particles under different gas temperatures was studied through cold spray experiments, and composite coatings were fabricated. The microstructure and phase composition were analyzed using scanning electron microscopy (SEM) and X-ray diffraction (XRD). The results showed that the numerical simulations were consistent with the experimental analyses. As the particle velocity or gas temperature increased, the degree of particle deformation upon deposition became more pronounced, accompanied by phenomena such as cracking or fragmentation and splashing rebound. At a gas temperature of 700 °C, both the bonding density of individual particles and the bonding effectiveness of multi-particle deposits were lower than those achieved at 500 °C. The coating prepared at a gas temperature of 500 °C exhibited a flatter surface, better overall bonding with the Ti interlayer, and higher internal density.