Fluorescent Confocal Microscopy
Fluorescent Confocal Microscopy - Related Videos
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,...
17:08
In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy
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Cited by 11 •
2014
Combinatorial 5 fluorescent proteins marking of hematopoietic stem and progenitor cells allows in vivo clonal tracking via confocal and two-photon microscopy, providing insights into bone marrow hematopoietic architecture during regeneration. This method allows non-invasive fate mapping of spectrally-coded HSPCs-derived cells in intact tissues for extensive periods of time following transplantation.
13:13
Confocal Fluorescence Microscopy: A Technique to Determine the Localization of Proteins in Mouse Fibroblasts
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2023
Source: Dominique R. Bollino1, Eric A. Legenzov2, Tonya J. Webb1 1 Department of Microbiology and Immunology, University of Maryland School of Medicine and the Marlene and Stewart Greenebaum Comprehensive Cancer Center, Baltimore, Maryland 21201 2 Center for Biomedical Engineering and Technology, University of Maryland School of Medicine, Baltimore, Maryland 21201 Confocal fluorescence microscopy is an imaging technique that enables increased optical resolution as compared to conventional...
09:44
Oral Biofilm Analysis of Palatal Expanders by Fluorescence In-Situ Hybridization and Confocal Laser Scanning Microscopy
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Cited by 20 •
2011
We present a protocol for structural and compositional analysis of natural oral biofilm from orthodontic appliances with in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM). Oral biofilm samples were collected from palatal expanders, scraping acrylic-resin flakes off their surface and referring them for molecular processing.
10:28
High-plex Imaging using Spectral Confocal Microscopy to Minimize Non-specific Tissue Fluorescence
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2025
Spectral Iterative Bleaching Extends Multiplexity (IBEX) builds upon the base IBEX technique by adding heparin blocking to minimize nonspecific binding and leveraging spectral detection with computational unmixing to suppress autofluorescence. This approach accelerates image acquisition while reducing sources of background, enabling robust multi-round, high-parameter spatial proteomic analyses.
FAQs
The focused laser excites fluorophores in a defined region of the specimen, allowing the instrument to detect selected fluorescent signals rather than treating the entire sample as uniformly visible. This selective excitation supports higher-contrast mapping of chemical distributions and helps distinguish molecular or material features within complex samples, including interfaces, polymers, and nanoparticles.
The pinhole rejects emission originating outside the focal region before detection. By reducing out-of-focus light, it enables collection of thin optical sections with improved depth resolution. These sections help reveal how fluorescently labeled structures or chemical signals are arranged at different positions within a specimen, rather than combining information from multiple depths into one image.
Fluorescent Confocal Microscopy collects optical sections from different depths within a specimen. The sections can then be assembled into a three-dimensional reconstruction that represents the spatial organization of detected fluorescent signals. In chemistry, this provides a way to examine molecular localization, material organization, or chemical distributions throughout a sample instead of observing only a single focal plane.
Fluorescent labels provide the detectable signals that connect a molecular or material feature to a measured image. Because the instrument records fluorescence from the labeled components, researchers can examine selected chemical distributions within complex samples. This selectivity is useful when the goal is to compare localization, organization, or concentration-related changes rather than simply view overall sample structure.
A typical workflow begins by associating the feature of interest with a fluorescent label, placing the specimen for optical imaging, and using a focused laser to excite the fluorophores. The instrument rejects out-of-focus emission through the pinhole, records thin sections at relevant depths, and can combine those sections into a three-dimensional representation for analysis.
Chemists can use the method when spatially resolved information is important, such as mapping molecular localization, monitoring a reaction over time, or characterizing the organization of polymers, nanoparticles, interfaces, or biological molecules. Repeated imaging can reveal changes in concentration or arrangement, while three-dimensional data can show how those changes vary through the sample depth.