Fluorescent proteins for optical microscopy

Author(s):  
Pingyong Xu ◽  
Mingshu Zhang ◽  
Hao Chang
Author(s):  
M. F. Garcia-Parajo ◽  
J.-A. Veerman ◽  
G. M. J. Segers-Nolten ◽  
B. G. de Grooth ◽  
J. Greve ◽  
...  

2009 ◽  
Vol 02 (01) ◽  
pp. 27-35 ◽  
Author(s):  
ADAM M. LARSON ◽  
ANTHONY LEE ◽  
PO-FENG LEE ◽  
KAYLA J. BAYLESS ◽  
ALVIN T. YEH

Ultrashort pulse, multispectral non-linear optical microscopy (NLOM) is developed and used to image, simultaneously, a mixed population of cells expressing different fluorescent protein mutants in a 3D tissue model of angiogenesis. Broadband, sub-10-fs pulses are used to excite multiple fluorescent proteins and generate second harmonic in collagen. A 16-channel multispectral detector is used to delineate the multiple non-linear optical signals, pixel by pixel, in NLOM. The ability to image multiple fluorescent protein mutants and collagen, enables serial measurements of cell-cell and cell-matrix interactions in our 3D tissue model and characterization of fundamental processes in angiogenic morphogenesis.


Author(s):  
D. J. Barber ◽  
R. G. Evans

Manganese (II) oxide, MnO, in common with CoO, NiO and FeO, possesses the NaCl structure and shows antiferromagnetism below its Neel point, Tn∼ 122 K. However, the defect chemistry of the four oxides is different and the magnetic structures are not identical. The non-stoichiometry in MnO2 small (∼2%) and below the Tn the spins lie in (111) planes. Previous work reported observations of magnetic features in CoO and NiO. The aim of our work was to find explanations for certain resonance results on antiferromagnetic MnO.Foils of single crystal MnO were prepared from shaped discs by dissolution in a mixture of HCl and HNO3. Optical microscopy revealed that the etch-pitted foils contained cruciform-shaped precipitates, often thick and proud of the surface but red-colored when optically transparent (MnO is green). Electron diffraction and probe microanalysis indicated that the precipitates were Mn2O3, in contrast with recent findings of Co3O4 in CoO.


Author(s):  
L. Montoto ◽  
M. Montoto ◽  
A. Bel-Lan

INTRODUCTION.- The physical properties of rock masses are greatly influenced by their internal discontinuities, like pores and fissures. So, these need to be measured as a basis for interpretation. To avoid the basic difficulties of measurement under optical microscopy and analogic image systems, the authors use S.E.M. and multiband digital image processing. In S.E.M., analog signal processing has been used to further image enhancement (1), but automatic information extraction can be achieved by simple digital processing of S.E.M. images (2). The use of multiband image would overcome difficulties such as artifacts introduced by the relative positions of sample and detector or the typicals encountered in optical microscopy.DIGITAL IMAGE PROCESSING.- The studied rock specimens were in the form of flat deformation-free surfaces observed under a Phillips SEM model 500. The SEM detector output signal was recorded in picture form in b&w negatives and digitized using a Perkin Elmer 1010 MP flat microdensitometer.


Author(s):  
John F. Mansfield

The current imaging trend in optical microscopy, scanning electron microscopy (SEM) or transmission electron microscopy (TEM) is to record all data digitally. Most manufacturers currently market digital acquisition systems with their microscope packages. The advantages of digital acquisition include: almost instant viewing of the data as a high-quaity positive image (a major benefit when compared to TEM images recorded onto film, where one must wait until after the microscope session to develop the images); the ability to readily quantify features in the images and measure intensities; and extremely compact storage (removable 5.25” storage devices which now can hold up to several gigabytes of data).The problem for many researchers, however, is that they have perfectly serviceable microscopes that they routinely use that have no digital imaging capabilities with little hope of purchasing a new instrument.


2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.


2006 ◽  
Vol 175 (4S) ◽  
pp. 328-328 ◽  
Author(s):  
Hugo H. Davila ◽  
Maggie Mamcarz ◽  
Irving Nadelhaft ◽  
Raoul Salup ◽  
Jorge Lockhart ◽  
...  

1996 ◽  
Vol 444 ◽  
Author(s):  
Maarten P. de Boer ◽  
Terry A. Michalske

AbstractWe have measured autoadhesion (e.g. stiction) of individual polysilicon beams by interferometric optical microscopy. Untreated cantilever beams were dried from water in air, while treated beams were coated with a hydrophobic molecular coating of octadecyltrichlorosilane (ODTS). Adhesion values obtained for beams adhered to the substrate over a long length (large d) are independent of beam length with values of 16.7 and 4.4 mJ/m2 for untreated and treated samples respectively. These values can be understood in terms of differences in surface chemistry and polysilicon roughness. Using the shortest length beam which remains attached to the substrate, adhesion values were 280 and 16 mJ/m2 respectively. These higher values may be a result of capillarity effects. We recommend that measurements be made on beams in which d is large, in contrast to the current practice of noting the shortest beam adhered.


2003 ◽  
Vol 773 ◽  
Author(s):  
Xiaohu Gao ◽  
Shuming Nie ◽  
Wallace H. Coulter

AbstractLuminescent quantum dots (QDs) are emerging as a new class of biological labels with unique properties and applications that are not available from traditional organic dyes and fluorescent proteins. Here we report new developments in using semiconductor quantum dots for quantitative imaging and spectroscopy of single cancer cells. We show that both live and fixed cells can be labeled with multicolor QDs, and that single cells can be analyzed by fluorescence imaging and wavelength-resolved spectroscopy. These results raise new possibilities in cancer imaging, molecular profiling, and disease staging.


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