Photoconvertible Fluorophores
Photoconvertible Fluorophores - Related Videos
07:59
Tracking Cells in GFP-transgenic Zebrafish Using the Photoconvertible PSmOrange System
0 Views •
Cited by 2 •
2016
We established the photoconvertible PSmOrange system as a powerful, straight-forward and cost inexpensive tool for in vivo cell tracking in GFP transgenic backgrounds. This protocol describes its application in the zebrafish model system.
07:19
Cell Tracking Using Photoconvertible Proteins During Zebrafish Development
0 Views •
Cited by 17 •
2012
Here, we present a method for the photoactivated switch of photoconvertible fluorescent proteins (PCFPs) in the living zebrafish embryo and further tracking of photoconverted protein at specific time points during development. This methodology allows monitoring of cell biological events underlying different developmental processes in a live vertebrate organism.
09:45
In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2
0 Views •
Cited by 7 •
2020
Presented here is a protocol to monitor degradation of the protein huntingtin fused to the photoconvertible fluorophore Dendra2.
14:11
Synthesis of pH Dependent Pyrazole, Imidazole, and Isoindolone Dipyrrinone Fluorophores using a Claisen-Schmidt Condensation Approach
0 Views •
2021
The Claisen-Schmidt condensation reaction is an important methodology for the generation of methine-bridged conjugated bicyclic aromatic compounds. Through utilizing a base-mediated variant of the aldol reaction, a range of fluorescent and/or biologically relevant molecules can be accessed through a generally inexpensive and operationally simple synthetic approach.
03:44
The Photoconversion Technique for Exploring Inflammatory Cell Dynamics in Insect Pupae
0 Views •
2025
The video demonstrates the use of photoconversion to study inflammatory cell dynamics in Drosophila pupae. Green hemocytes are drawn to the wounded epithelium, followed by distant migration. Photoconversion shifts certain hemocytes from green to red, facilitating their movement tracking.
FAQs
Focused illumination at a specific wavelength triggers a photochemical rearrangement within the fluorescent molecule. This rearrangement changes the probe’s excitation or emission spectrum, so the illuminated region can be distinguished from surrounding labeled material. Because the change occurs only where the appropriate light is applied, researchers can mark selected cells, organelles, or proteins within a larger population.
A permanent change preserves the identity of the illuminated population as imaging continues. Researchers can therefore compare the marked material with its later position or distribution, rather than relying only on a transient observation. In neuronal studies, this supports analysis of how labeled structures or molecules move through axons and how neuronal architecture changes over time.
A spectral shift provides a way to distinguish converted material from unconverted material in the same sample. This creates a spatial and temporal reference for following selected structures while other labeled material remains visible. The approach is particularly useful when researchers need to determine where a marked molecule, organelle, or cellular region was located and how its distribution subsequently changes.
Photoconversion adds a spatially selective labeling step to broader imaging and circuit-mapping approaches. It can identify a chosen subset of cells or structures and then relate their later behavior to neuronal architecture or activity-related structural changes. Thus, the technique complements methods that map connectivity or activity by providing a way to track selected material across time.
Researchers first use the probe to label the cells, organelles, or proteins of interest and document their initial fluorescence. They then apply focused illumination at the wavelength that induces conversion in a selected region. Subsequent imaging distinguishes the altered signal from the original state, allowing changes in location, neuronal structure, or distribution to be followed over time.
The converted population can be used to examine neuronal architecture, axonal transport, and synaptic remodeling. Tracking the altered signal helps reveal how selected material is distributed or relocated after labeling. The same strategy also supports cell-lineage studies, where marked cells can be distinguished from unconverted populations as observations continue.
Photoconvertible fluorophores are valuable when a study must connect a defined starting location with later structural or transport behavior. Applications include live imaging of neuronal architecture, monitoring material moving through axons, examining synaptic remodeling, and following cell lineage. They also provide context for circuit mapping and for investigating structural changes associated with neuronal activity.