Real-time Fluorescence Tracing
Real-time Fluorescence Tracing - Related Videos
06:37
Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology
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Cited by 41 •
2010
Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.
12:23
Real-time Monitoring of Ligand-receptor Interactions with Fluorescence Resonance Energy Transfer
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Cited by 6 •
2012
We demonstrate FRET between conjugated polymer polydiacetylene (PDA) and fluorophore attached to the surface of PDA liposomes for the sensing of biomolecules. PDA liposomes also contained receptor molecules on their surfaces for biomolecules to be used as probes. Ligand-receptor interactions lead to changes in the FRET efficiency between the fluorophore and PDA which is the basis of the sensing mechanism.
08:32
Application of Genetically Encoded Fluorescent Nitric Oxide (NO•) Probes, the geNOps, for Real-time Imaging of NO• Signals in Single Cells
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Cited by 21 •
2017
This manuscript presents protocols for the application of novel genetically encoded nitric oxide (NO•) probes (geNOps) to monitor single cell NO• fluctuations in real-time using fluorescence microscopy. The Ca2+-triggered NO• formation on the level of individual endothelial cells was visualized by combining geNOps with a chemical Ca2+ sensor.
07:48
A Method for Lineage Tracing of Corneal Cells Using Multi-color Fluorescent Reporter Mice
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Cited by 15 •
2015
In this article we describe the principles for designing and performing a multi-color lineage tracing experiment using R26R-Confetti mice. We provide a specific protocol for tracking corneal epithelial cells which can be modified for other tissues of interest.
08:21
Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor
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Cited by 9 •
2017
This protocol outlines a simple method for analyzing calcium signals in plants generated by feeding hemipteran insects, such as aphids. Arabidopsis thaliana transformed with the GFP calcium biosensor GCaMP3 allow for the real-time in vivo imaging of calcium dynamics with a high temporal and spatial resolution.
FAQs
The method relies on a two-stage optical interaction. A fluorescent probe is excited with light at a specific wavelength, and the emitted light is then detected repeatedly over time. This sequence converts molecular or cellular behavior into a time-resolved signal, allowing investigators to follow changes as they happen rather than infer them from a single measurement.
Each signal dimension can provide a different type of biological information. Changes in intensity may reflect altered activity, while shifts in location can indicate movement or uptake. Fluorescence lifetime, meaning how long the signal persists after excitation, can report changes in the probe’s local environment. Together, these measurements help distinguish dynamic biological events.
Static measurements capture a biological state at one point, whereas real-time fluorescence tracing preserves the timing of change. That temporal information can show whether a labeled molecule moves, binds, or is taken up, and can connect those events with changing cellular or tissue conditions. The result is a more continuous view of biological activity.
A basic workflow begins by associating a fluorescent label with the molecule, cell, or biological event of interest. The labeled target is exposed to light at the probe’s excitation wavelength, and emitted fluorescence is recorded over time. Investigators then examine changes in signal intensity, position, or lifetime to interpret movement, uptake, binding, or environmental changes.
Medical researchers can apply real-time fluorescence tracing to follow drug delivery, observe cellular behavior, examine tissue function, or monitor disease-associated processes. Because the method records changes as they occur, it can help relate the location or activity of a labeled target to evolving biological conditions, supporting investigations that require more than a single endpoint measurement.
By linking molecular activity with changing physiology, the method can provide information relevant to diagnosis, treatment response, and targeted therapies. Tracking labeled molecules or cells may reveal how a process develops over time and where activity occurs. In medical research, this connection can help investigators study disease mechanisms and evaluate how biological systems respond to treatment.