Epi-fluorescence Imaging
Epi-fluorescence Imaging - Related Videos
02:13
In Vivo Imaging of Geniculate Ganglion Neuron Responses Using Epifluorescent Microscopy
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2025
Source: Fowler, B. E., et al. In vivo Calcium Imaging of Mouse Geniculate Ganglion Neuron Responses to Taste Stimuli. J. Vis. Exp. (2021) This video demonstrates a protocol for delivering tastants to the surgically exposed geniculate ganglion in an anesthetized mouse and using GCaMP fluorescence with epifluorescent microscopy to visualize real-time neuronal responses, aiding studies on peripheral taste signal processing.
08:32
Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software
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Cited by 8 •
2011
The C. elegans embryo is a powerful system for studying cell biology and development. We present a protocol for live imaging of C. elegans embryos utilizing DIC optics or fluorescence using readily available epifluorescent microscopes and open-source software.
09:00
Long-term Intravital Immunofluorescence Imaging of Tissue Matrix Components with Epifluorescence and Two-photon Microscopy
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Cited by 21 •
2014
The extracellular matrix undergoes substantial remodeling during wound healing, inflammation and tumorigenesis. We present a novel intravital immunofluorescence microscopy approach to visualize the dynamics of fibrillar as well as mesh-like matrix components with high spatial and temporal resolution using epifluorescence or two-photon microscopy.
06:31
Real-time Imaging of Plant Cell Surface Dynamics with Variable-angle Epifluorescence Microscopy
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Cited by 10 •
2015
The goal of this protocol is to demonstrate how to monitor fluorescently-tagged protein dynamics on plant cell surfaces with variable-angle epifluorescence microscopy, showing blinking dots of GFP-tagged PATROL1, a membrane trafficking protein, in the cell cortex of the stomatal complex in Arabidopsis thaliana.
07:44
Time-Lapse Epifluorescence Microscopy Imaging of Pseudomonas aeruginosa and Staphylococcus aureus Heterogeneous Phenotypes
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Cited by 2 •
2025
In this manuscript, we provide a comprehensive protocol for assessing antibiotic survival for Pseudomonas aeruginosa and Staphylococcus aureus, transforming plasmids into P. aeruginosa and S. aureus to create reporter strains and visualize phenotypic variants, such as persisters, by time-lapse epifluorescence microscopy.
FAQs
The objective directs excitation light onto the specimen, where fluorophores absorb selected wavelengths. These molecules then emit light at longer wavelengths. Because the illumination and emitted signal differ spectrally, the microscope can collect the fluorescence associated with labeled structures rather than treating the incoming excitation as the image signal. This enables visualization of molecular features within biological specimens.
The dichroic mirror and emission filter separate the fluorescence produced by the specimen from the excitation light used to stimulate it. This separation is essential because the detector must receive the emitted signal associated with labeled structures while limiting the contribution of the illumination path. Their coordinated action supports imaging of cellular and tissue features marked by fluorophores.
Compatibility with multiple fluorescent labels allows researchers to examine more than one labeled feature within a biological specimen. Because fluorophores respond to specific excitation wavelengths and emit longer-wavelength light, different labels can support observations of distinct molecular or cellular features. This expands the ability of Epi-fluorescence Imaging to investigate organization within cells and tissues.
Fluorescent labels associate the observed signal with selected structures, such as proteins or organelles. The resulting image therefore provides information about where those labeled components are positioned within cells, tissues, or other specimens. In addition to showing overall morphology, this approach makes molecular organization observable and helps connect structural patterns with cellular behavior.
Researchers can use the method to localize proteins, examine organelles, and monitor cell morphology. These readouts connect fluorescent signals with both molecular placement and visible changes in cellular form. The same imaging approach can also make cellular behavior and biological changes observable, allowing studies to address structure and dynamics rather than relying on morphology alone.
It is useful when researchers need to monitor biological changes in live samples or compare cellular states across time. Fluorescently labeled structures can be followed while cell morphology or behavior changes, providing a way to relate molecular organization to dynamic observations. This makes the technique relevant to live-sample analysis and studies of changing cellular conditions.
The approach is valuable across cell biology, developmental biology, pathology, and live-sample analysis. In cell biology, it can support protein and organelle localization; in developmental biology, it can reveal labeled structures as biological patterns change. Pathology applications can examine labeled features in specimens, while live-sample studies can monitor morphology or other changes over time.
Epi-fluorescence Imaging can be applied to cells, tissues, and other biological specimens that contain fluorescently labeled structures. This range supports questions at different biological scales, from the placement of proteins or organelles within individual cells to labeled features distributed through tissue. The appropriate specimen depends on whether the study focuses on organization, morphology, or biological change.