222,848 research outputs found
Confocal microscopy
Chapter focusing on confocal microscopy. A confocal microscope is one in which the illumination is confined to a small volume in the specimen, the detection is confined to the same volume and the image is built up by scanning this volume over the specimen, either by moving the beam of light over the specimen or by displacing the specimen relative to a stationary beam. The chief advantage of this type of microscope is that it gives a greatly enhanced discrimination of depth relative to conventional microscopes. Commercial systems appeared in the 1980s and, despite their high cost, the world market for them is probably between 500 and 1000 instruments per annum, mainly because of their use in biomedical research in conjunction with fluorescent labelling methods. There are many books and review articles on this subject ( e.g. Pawley ( 2006) , Matsumoto( 2002), Wilson (1990) ). The purpose of this chapter is to provide an introduction to optical and engineering aspects that may be o f interest to biomedical users of confocal microscopy
Twin-Photon Confocal Microscopy
A recently introduced two-channel confocal microscope with correlated
detection promises up to 50% improvement in transverse spatial resolution
[Simon, Sergienko, Optics Express {\bf 18}, 9765 (2010)] via the use of photon
correlations. Here we achieve similar results in a different manner,
introducing a triple-confocal correlated microscope which exploits the
correlations present in optical parametric amplifiers. It is based on tight
focusing of pump radiation onto a thin sample positioned in front of a
nonlinear crystal, followed by coincidence detection of signal and idler
photons, each focused onto a pinhole. This approach offers further resolution
enhancement in confocal microscopy
Improving spatial resolution of confocal Raman microscopy by super-resolution image restoration
A new super-resolution image restoration confocal Raman microscopy method (SRIR-RAMAN) is proposed for improving the spatial resolution of confocal Raman microscopy. This method can recover the lost high spatial frequency of the confocal Raman microscopy by using Poisson-MAP super-resolution imaging restoration, thereby improving the spatial resolution of confocal Raman microscopy and realizing its super-resolution imaging. Simulation analyses and experimental results indicate that the spatial resolution of SRIR-RAMAN can be improved by 65% to achieve 200 nm with the same confocal Raman microscopy system. This method can provide a new tool for high spatial resolution micro-probe structure detection in physical chemistry, materials science, biomedical science and other areas
Use of confocal and multiphoton microscopy for the evaluation of micro-optical components and emitters
We report on the application of confocal and multiphoton microscopic techniques for the evaluation of the latest generation of micro optical components. The optical emitting characteristics of arrays of matrix addressable GaN micrometer-sized light emitting diodes (micro-LEDs) have been measured using a commercial confocal microscope utilising the LEDs' own emission along with reflection confocal microscopy to determine the surface structure. Multiphoton induced luminescence from the 10-20-micron diameter emitters has also been used to examine the structure of the device and we compare this with electrically induced emission. In related work, the optical properties of micro lens arrays (10-100-micron diameter) fabricated in SiC, Sapphire, and Diamond have been determined using transmission confocal microscopy. Such optical microscopy techniques offer a simple, non-destructive method to determine the structure and performance of such novel devices
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Rifabutin corneal deposits localized to the deep stroma using anterior segment optical coherence tomography.
Purpose:To demonstrate that rifabutin-related corneal deposits are localized to the deep stroma using anterior segment optical coherence tomography (OCT) and confocal microscopy. Observations:A 55-year-old male with a history of human immunodeficiency virus (HIV) and disseminated mycobacterium avium complex on rifabutin treatment for 3 years presented with bilateral corneal deposits. Confocal microscopy and anterior segment OCT confirm that rifabutin-related corneal deposits are located in the deep stroma, rather than in the endothelium. Conclusions:And Importance: Rifabutin deposits localize to the deep corneal stroma, and can be seen with both confocal microscopy and anterior segment OCT. Anterior segment OCT is a widely available and easily used diagnostic tool, and can provide utility in the diagnosis of corneal deposits
Methods to calibrate and scale axial distances in confocal microscopy as a function of refractive index
Accurate distance measurement in 3D confocal microscopy is important for
quantitative analysis, volume visualization and image restoration. However,
axial distances can be distorted by both the point spread function and by a
refractive-index mismatch between the sample and immersion liquid, which are
difficult to separate. Additionally, accurate calibration of the axial
distances in confocal microscopy remains cumbersome, although several high-end
methods exist. In this paper we present two methods to calibrate axial
distances in 3D confocal microscopy that are both accurate and easily
implemented. With these methods, we measured axial scaling factors as a
function of refractive-index mismatch for high-aperture confocal microscopy
imaging. We found that our scaling factors are almost completely linearly
dependent on refractive index and that they were in good agreement with
theoretical predictions that take the full vectorial properties of light into
account. There was however a strong deviation with the theoretical predictions
using (high-angle) geometrical optics, which predict much lower scaling
factors. As an illustration, we measured the point-spread-function of a
point-scanning confocal microscope and showed that an index-matched,
micron-sized spherical object is still significantly elongated due to this PSF,
which confirms that single micron-sized spheres are not well suited to
determine accurate axial calibration nor axial scaling.Comment: 8 pages, 5 figure
Confocal laser scanning microscope, raman microscopy and western blotting to evaluate inflammatory response after myocardial infarction
Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead
cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the
inflammatory phase, the proliferative phase and the maturation phase.
Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory
response following myocardial infarction both humoral and cellular phase, allowing the identification and
semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution
and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy
(CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the
sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific
protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced
semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution
of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy.
The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample
and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated
the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this
way additional information regarding the extension of the inflammatory process in function of the time stayed from the
time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory
mediators at several different timepoints providing a more detailed analysis of the time course of the infarct
Plane-projection multi-photon microscopy for high-frame-rate Live Tissue Imaging
We present a wide-field multi-photon microscopy that provides optical sectioning at high frame rate under biocompatible laser dosage. Axial resolution comparable to confocal microscopy and 5-frame-per-second live tissue imaging are demonstrated
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