Fluorescence Microscopy Assay
Fluorescence Microscopy Assay - Related Videos
10:27
A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae
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Cited by 26 •
2011
A robust approach to monitor the delivery of organelles to the acidic lumen of the yeast vacuole for degradation and recycling is described. The method relies on the specific labeling of target organelles with a genetically encoded dual-emission fluorescence pH-biosensor, and visualization of individual cells using fluorescence microscopy.
09:22
Introduction to Fluorescence Microscopy
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2023
Fluorescence microscopy is a very powerful analytical tool that combines the magnifying properties of light microscopy with visualization of fluorescence. Fluorescence is a phenomenon that involves absorbance and emission of a small range of light wavelengths by a fluorescent molecule known as a fluorophore. Fluorescence microscopy is accomplished in conjunction with the basic light microscope by the addition of a powerful light source, specialized filters, and a means of fluorescently...
11:23
Lensless Fluorescent Microscopy on a Chip
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Cited by 8 •
2011
A lensless on-chip fluorescent microscopy platform is demonstrated that can image fluorescent objects over an ultra-wide field-of-view of e.g., >0.6-8 cm2 with <4μm resolution using a compressive sampling based decoding algorithm. Such a compact and wide-field fluorescent on-chip imaging modality could be valuable for high-throughput cytometry, rare-cell research and microarray-analysis.
10:07
"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)
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Cited by 8 •
2016
We describe an experimental setup to visualize with unprecedented high resolution phagosome formation and closure in three dimensions in living macrophages, using total internal reflection fluorescence microscopy. It allows monitoring of the base of the phagocytic cup, the extending pseudopods, as well as the precise site of phagosome scission.
09:02
Visualizing Bacteria in Nematodes using Fluorescent Microscopy
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Cited by 20 •
2012
To study the mutualism between Xenorhabdus bacteria and Steinernema nematodes, methods were developed to monitor bacterial presence and location within nematodes. The experimental approach, which can be applied to other systems, entails engineering bacteria to express the green fluorescent protein and visualizing, using fluorescence microscopy bacteria within the transparent nematode.
FAQs
The fluorophore first absorbs excitation light and then emits light at a longer wavelength. Optical filters separate this emitted signal from the original illumination before it reaches the detector. This separation allows the recorded image to represent label-associated fluorescence rather than the excitation source, supporting spatially resolved observation of biological structures or events.
Selective labeling connects a fluorescent signal to a particular molecule, cell population, or biological event. Image analysis can then relate that signal to cellular structure and function, rather than treating all visible features as equivalent. This principle supports measurements such as protein localization, gene expression, molecular interactions, and cell viability within a defined biological context.
The assay can be applied to either fixed or living specimens, allowing researchers to address different biological questions. Fixed samples support examination of labeled features in a preserved specimen, whereas living samples can support investigation of dynamic processes. This distinction helps determine whether an experiment emphasizes spatial organization, ongoing cellular behavior, or both.
A typical workflow begins by applying fluorescent labels to the molecules, cells, or events of interest. The specimen is then illuminated with excitation light, while emitted light passes through optical filters to a detector. The resulting image or signal undergoes analysis to visualize or quantify the labeled target and relate it to cellular organization.
Fluorescence microscopy assay data can provide spatial information about protein localization and measurements related to gene expression, cell viability, or molecular interactions. Image analysis converts label-associated patterns into interpretable results, allowing researchers to connect molecular identity with cellular structure. The resulting measurements can reveal how biological components are organized or how events change in an experiment.
Biology researchers use fluorescence microscopy assays in experimental cell biology, disease-mechanism studies, and drug-response investigations. The method is especially useful when a study must connect a labeled molecular target with its cellular location or behavior. Depending on the specimen and labeling strategy, it can support analysis of organization, viability, interactions, gene expression, or dynamic cellular processes.