Confocal Raman Microscopy
Confocal Raman Microscopy - Related Videos
09:57
Multiplex Chemical Imaging Based on Broadband Stimulated Raman Scattering Microscopy
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Cited by 8 •
2022
We present a protocol to acquire chemical images with broadband stimulated Raman scattering (SRS) microscopy. Based on an SRS microscope that operates with differential multichannel-lock-in detection, the protocol describes the sample preparation, adjustment of the SRS apparatus, and chemometrics to disentangle different constituents of chemically heterogeneous samples.
10:48
Imaging Biological Samples with Optical and Confocal Microscopy
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2023
Source: Peiman Shahbeigi-Roodposhti and Sina Shahbazmohamadi, Biomedical Engineering Department, University of Connecticut, Storrs, Connecticut Optical microscopes have been around for centuries, and while they reached their theoretical limitation of resolution decades ago, new equipment and techniques, such as confocal and digital image processing, have created new niches within the field of optical imaging. The best optical microscopes will typically have a resolution down to 200 nm in ideal...
03:16
Three-dimensional Imaging of Immunolabeled Astrocytes Using Confocal Microscopy
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2025
Source: Bagheri, M., et. al. Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes. J. Vis. Exp. (2015).This video demonstrates the imaging of fluorescently labeled astrocytes in a mouse brain section using a confocal microscope. The brain section is positioned under the appropriate objective lens, and suitable fluorophores and filters are selected. After optimizing the exposure automatically, the scanning parameters are defined to ensure high-resolution 2D images. Z-stack...
01:16
Confocal Fluorescence Microscopy
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2023
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
09:16
Analyzing Craniofacial Morphogenesis in Zebrafish Using 4D Confocal Microscopy
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Cited by 18 •
2014
Time-lapse confocal imaging is a powerful technique useful for characterizing embryonic development. Here, we describe the methodology and characterize craniofacial morphogenesis in wild-type, as well as pdgfra, smad5, and smo mutant embryos.
FAQs
The laser interacts with the illuminated sample volume, producing inelastically scattered light whose wavelength shifts correspond to molecular vibrations. Because different constituents produce different vibrational signatures, the resulting spectrum can indicate phase composition and help distinguish materials within polymers, semiconductors, ceramics, and composites. This chemical information supports engineering characterization without requiring invasive sampling.
The confocal pinhole suppresses light originating outside the focal region before detection. This improves depth discrimination and spatial resolution, allowing measurements to be associated with specific locations inside or beneath a material surface. By changing the sampled position through the material, engineers can examine composition, interfaces, defects, and other features across three dimensions rather than only at the surface.
Raman measurements can reveal variations in phase composition, stress and strain, defects, and interfaces. These signals allow engineers to connect local chemical or structural differences with material performance and processing history. The approach is especially useful when a component contains multiple constituents or when a failure may originate from a small, spatially localized region.
A focused laser is directed to selected locations within the sample, and the instrument records the wavelength-shifted scattered light from each focal volume. Measurements collected across positions can then be compared to identify spatial changes in composition or material condition. Confocal rejection of out-of-focus signals improves the reliability of location-specific data during the resulting map.
Engineers apply the technique when they need chemical and spatial information from complex materials without physically removing or sectioning the sample. It can characterize constituents, phase distributions, interfaces, and localized defects in polymers, ceramics, and composites. Those results are relevant to material selection, performance evaluation, and investigations in which internal heterogeneity affects reliability.
Spatially resolved spectra can help compare intact and suspect regions, locate compositional differences, and examine defects or interfaces associated with a failed component. The same information can support quality control by checking material consistency and identifying changes during processing. In advanced device development, mapping also helps evaluate material condition in structures where noninvasive analysis is valuable.