Live Fluorescent Probes
Live Fluorescent Probes - Related Videos
11:21
Photoconversion of Purified Fluorescent Proteins and Dual-probe Optical Highlighting in Live Cells
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Cited by 5 •
2010
This protocol describes a general approach to perform photoconversion of fluorescent proteins on a confocal laser scanning microscope. We describe procedures for the photoconversion of puried protein samples, as well as for dual-probe optical highlighting in live cells with mOrange2 and Dronpa.
10:59
Determination of Lipid Raft Partitioning of Fluorescently-tagged Probes in Living Cells by Fluorescence Correlation Spectroscopy (FCS)
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Cited by 9 •
2012
A technique to probe the lipid raft partitioning of fluorescent proteins at the plasma membrane of living cells is described. It takes advantage of the disparity in diffusion times of proteins located inside or outside of lipid rafts. Acquisition can be performed dynamically in control conditions or after drug addition.
12:19
Rapid Subtractive Patterning of Live Cell Layers with a Microfluidic Probe
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Cited by 2 •
2016
We present a protocol to perform subtractive patterning of live cell monolayers on a surface. This is achieved by local and selective lysis of adherent cells using a microfluidic probe (MFP). The cell lysate retrieved from local regions can be used for downstream analysis, enabling molecular profiling studies.
04:02
An Assay to Measure the Accumulation of a Fluorescent Antibiotic Probe in Bacterial Cells
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2025
This video demonstrates an assay to measure the accumulation of a fluorescent antibiotic probe in bacterial cells. Exposure to an efflux pump inhibitor disrupts the transmembrane proton gradient in bacterial cells, upon which a fluorophore-conjugated antibiotic is added. Inhibition of efflux pumps due to the disrupted proton gradient causes accumulation of the antibiotic inside the bacteria. Bacterial cells are lysed, and the fluorescence intensity is measured to confirm antibiotic accumulation.
09:45
Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
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Cited by 17 •
2012
Fluorescence Lifetime Imaging (FLIM) has emerged as a key technique to image the environment and interaction of specific proteins and dyes in living cells. FLIM of fluorescent molecular rotors allows mapping of viscosity in living cells.
FAQs
The probe links a biological response to a measurable change in fluorescence. Depending on its design, the fluorophore can become brighter or dimmer, shift its emission wavelength, or change location within the cell. These signal changes correspond to the presence or activity of a target molecule, ion, enzyme, or cellular condition, allowing dynamic events to be monitored optically.
Each readout provides a different type of information about cellular behavior. Brightness changes can indicate altered probe response, wavelength shifts can distinguish changes in the optical signal, and localization can show where a relevant event occurs. Considering these dimensions together helps researchers connect molecular or cellular activity with both its intensity and spatial distribution.
Live fluorescent probes preserve access to information about when and where a biological event occurs, whereas endpoint measurements provide an observation after the process has reached a selected stage. This distinction matters when signaling, metabolism, gene expression, or cellular responses change over time. In bioengineering, temporal information can reveal dynamics that a single final measurement may miss.
Selection begins with the biological event that must be monitored, such as signaling, metabolism, gene expression, an ion-related response, enzyme activity, or a cellular condition. Researchers then consider whether brightness, wavelength, or localization best reports that event. Matching the probe response to the intended measurement helps produce information that is relevant to the engineered cell or system.
They can be used to examine dynamic signaling, metabolic behavior, gene expression, and interactions between cells and biomaterials. These applications extend beyond simply detecting whether an event occurred. By preserving spatial and temporal information, the measurements can help researchers evaluate how engineered cells behave and how biomaterial environments influence cellular processes over time.
Real-time optical measurements can help researchers assess engineered cells and follow biological processes relevant to therapeutic design. Monitoring signaling, metabolism, gene expression, or cellular responses provides evidence about system behavior while the sample remains observable. That information can guide evaluation and optimization of therapeutic systems, particularly when dynamic changes are important but endpoint measurements provide insufficient context.