Correlated Light Microscopy
Correlated Light Microscopy - Related Videos
11:16
Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Cited by 12 •
2016
A method is described whereby quantum dot (QD) nanoparticles can be used for correlative immunocytochemical studies of epoxy embedded human pathology tissue. We employ commercial antibody fragment conjugated QDs that are visualized by widefield fluorescence light microscopy and transmission electron microscopy.
10:00
Low-Cost Cryo-Light Microscopy Stage Fabrication for Correlated Light/Electron Microscopy
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Cited by 11 •
2011
We demonstrate the fabrication of a low-cost cryogenic stage designed to fit most reflected light microscopes. This lab-built cryogenic stage enables efficient and reliable correlative imaging between cryo-light and cryo-electron microscopy.
10:52
Correlative Light and Electron Microscopy to Study Microglial Interactions with β-Amyloid Plaques
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Cited by 30 •
2016
This article describes a protocol for visualizing amyloid Aβ plaques in Alzheimer's disease mouse models using methoxy-X04, which crosses the blood-brain barrier and selectively binds to β-pleated sheets found in dense core Aβ plaques. It allows pre-screening of plaque-containing tissue sections prior to immunostaining and processing for electron microscopy.
08:47
Epon Post Embedding Correlative Light and Electron Microscopy
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Cited by 1 •
2024
We present a detailed protocol for Epon post-embedding correlative light and electron microscopy using a fluorescent protein called mScarlet. This method can maintain the fluorescence and the ultrastructure simultaneously. This technique is amenable to a wide variety of biological applications.
09:10
Correlative Light and Electron Microscopy (CLEM) as a Tool to Visualize Microinjected Molecules and their Eukaryotic Sub-cellular Targets
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Cited by 19 •
2012
The CLEM technique has been adapted to analyze ultrastructural morphology of membranes, organelles, and subcellular structures affected by microinjected molecules. This method combines the powerful techniques of micromanipulation/microinjection, confocal fluorescent microscopy, and electron microscopy to allow millimeter to multi-nanometer resolution. This technique is amenable to a wide variety of applications.
FAQs
Spatial registration aligns the light-m microscopy observation with a complementary image by using shared landmarks in the specimen. This alignment allows a fluorescent signal associated with a molecule or organelle to be interpreted within the corresponding cellular architecture. Without reliable spatial matching, the two datasets cannot be confidently related, limiting conclusions about molecular location and structural context.
Light microscopy provides molecularly specific information through fluorescent labeling and can capture cellular observations that complement structural imaging. Electron microscopy contributes higher-resolution ultrastructural context. Combining these views helps investigators connect a labeled protein or organelle with fine cellular architecture rather than interpreting fluorescence without structural reference or structure without molecular identification.
Fluorescent probes identify specific proteins or organelles within the specimen, giving the light-microscopy dataset chemical or molecular specificity. Their signals can then be compared with structural features in a complementary image. This pairing helps determine where a target is located and supports interpretation of its relationship to cellular organization and function.
Interpretation depends on retaining shared landmarks and achieving accurate spatial registration between the imaging datasets. The labeled target must also produce a light-microscopy signal that can be related to the specimen's structural features. When these relationships are preserved, researchers can connect molecular localization with architecture; when they are unclear, biological conclusions become more limited.
A typical workflow begins by preparing a specimen with fluorescent labels for selected proteins or organelles. Researchers first acquire the light-microscopy observation, then obtain complementary structural images or measurements. They align the datasets through shared landmarks and spatial registration, allowing the fluorescent observations to be examined in relation to cellular ultrastructure and organization.
The approach is most valuable when a single imaging method cannot provide molecular specificity, structural detail, and functional context at the same time. In biology, it supports studies of infection, intracellular trafficking, and organelle organization. The combined datasets help investigators relate labeled components to the architecture in which cellular processes occur.
Correlated light microscopy can help localize molecules, identify their association with organelles or other cellular structures, and interpret cellular architecture in functional terms. By connecting chemically specific or dynamic observations with ultrastructural context, it supports analysis of how labeled components are organized and how processes such as trafficking or infection relate to cell structure.