Confocal Microscopy
Confocal Microscopy - Related Videos
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...
09:20
Near Simultaneous Laser Scanning Confocal and Atomic Force Microscopy (Conpokal) on Live Cells
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Cited by 10 •
2020
Presented here is the protocol of the Conpokal technique, which combines confocal and atomic force microscopy into a single instrument platform. Conpokal provides same cell, same region, near simultaneous confocal imaging and mechanical characterization of live biological samples.
09:14
Visualizing Macrophage Extracellular Traps Using Confocal Microscopy
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Cited by 18 •
2017
Macrophage extracellular traps are a newly described entity. This article will concentrate on confocal microscopy methods and how they are visualized in vitro and in vivo from lung samples.
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,...
FAQs
The laser illuminates a focused location rather than the entire specimen at once, and the detector records the emitted fluorescence from that scanned point. Repeating this process across the field builds an image from localized measurements. This coordinated scanning approach helps preserve spatial detail when researchers examine cellular structures or subcellular components within complex biological samples.
The pinhole sits in front of the detector and blocks fluorescence arriving from regions outside the selected focal plane. Because light from the focused location can pass to the detector while much out-of-focus emission is excluded, each recorded optical section contains more spatially restricted information. This enables clearer analysis of structures positioned at different depths in a specimen.
Fluorescent labels provide the emitted signal that the microscope detects during scanning, allowing particular biological structures or proteins to be examined within their spatial context. The resulting images can reveal protein localization, cell morphology, and relationships among subcellular components. For living specimens, the probes must also remain compatible with the imaging conditions used during observation.
Researchers first prepare a specimen containing suitable fluorescently labeled structures, then scan selected regions point by point. They collect sequential optical sections at different positions through the sample rather than relying on a single image. Combining those sections produces a three-dimensional reconstruction that can show how cells, tissues, or subcellular components are organized across depth.
This approach is useful when the research question depends on spatial organization within cells or tissues. Images can support examination of cell morphology, localization of proteins, and the arrangement of subcellular components in three dimensions. The method therefore connects molecular labeling with structural information, helping investigators determine where particular features occur within a biological specimen.
Living-specimen imaging is possible when the fluorescent probes and imaging conditions are compatible with continued observation. Under those circumstances, sequential images can be used to examine dynamic biological processes rather than only fixed structural relationships. The choice of labeling and imaging conditions is therefore important because it determines whether the specimen can be monitored while retaining interpretable fluorescence-based information.