Epifluorescence Stereomicroscopy
Epifluorescence Stereomicroscopy - 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.
14:29
Capturing Tissue Repair in Zebrafish Larvae with Time-lapse Brightfield Stereomicroscopy
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Cited by 27 •
2015
We present a protocol for capturing the dynamics of zebrafish larval tail fin regeneration on a whole-tissue scale using brightfield-based stereomicroscopy. This technique enables capturing the regeneration dynamics with single cell resolution. This methodology can be adapted to any stereomicroscope equipped with a CCD camera and time-lapse software.
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.
06:27
Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence
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Cited by 1 •
2019
We present the protocols to examine mouse heart development using whole mount epifluorescent microscopy on mouse embryos dissected from ventricular specific MLC-2v-tdTomato reporter knock-in mice. This method allows us to directly visualize each stage of the ventricular formation during mouse heart development without labor-intensive histochemical methods.
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.
FAQs
The excitation source sends light through the objective toward the specimen. Fluorescence generated by labeled structures returns through the optical system, where a dichroic mirror and emission filter separate it from the excitation light. The remaining signal reaches the camera or eyepieces, allowing the labeled features to be observed with reduced interference from the illumination.
Depth perception comes from the stereomicroscope’s three-dimensional view, while low magnification preserves a broader view of the specimen. This combination helps users relate fluorescently detected structures to the specimen’s surrounding form and supports natural handling during observation. It is particularly useful when positioning, dissecting, or manipulating biological material under direct visual control.
Selective contrast comes from detecting light emitted by fluorescent labels rather than relying only on the specimen’s overall appearance. Structures carrying labels, including fluorescently tagged proteins, can therefore stand out against surrounding biological material. This makes it easier to locate specific features within embryos, insects, tissues, cell cultures, or other specimens being examined.
The objective places excitation light onto the specimen. The resulting fluorescence then encounters the dichroic mirror and emission filter, which separate the emitted signal from excitation light before transmission to the camera or eyepieces. This arrangement links illumination with selective detection, allowing labeled structures to be viewed rather than simply recording the incoming excitation.
A basic observation sequence begins with placing a labeled biological specimen for viewing and directing excitation light through the objective. The emitted fluorescence is separated from excitation light by the dichroic mirror and emission filter, then viewed through eyepieces or recorded with a camera. This workflow can also support real-time examination of living samples.
The technique is suited to specimens in which fluorescent structures must be located within a broader three-dimensional form. Applications described for it include observing embryos, insects, tissues, and cell cultures, as well as examining fluorescently tagged proteins. Its combination of selective contrast, depth perception, and specimen handling supports varied biological sample types.
In developmental biology, the method can provide real-time visual information from living samples while preserving depth perception and natural handling. The same capabilities support dissection, microinjection, and sorting, where users need to see labeled structures and manipulate specimens directly. These applications connect fluorescence-based localization with practical examination or handling of biological material.