Fluorescent Probes
Fluorescent Probes - Related Videos
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.
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.
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.
07:14
Assessing Phagocytic Clearance of Cell Death in Experimental Stroke by Ligatable Fluorescent Probes
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Cited by 4 •
2014
We present a new fluorescence technique for selective in situ labeling of active phagocytic cells, which clear off cell corpses in stroke. The approach is important for assessing brain reaction to ischemia because only a small proportion of phagocytes present in ischemic brain participate in clearance of cell death.
08:23
Visualization of Bacterial Resistance using Fluorescent Antibiotic Probes
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Cited by 8 •
2020
Fluorescently tagged antibiotics are powerful tools that can be used to study multiple aspects of antimicrobial resistance. This article describes the preparation of fluorescently tagged antibiotics and their application to studying antibiotic resistance in bacteria. Probes can be used to study mechanisms of bacterial resistance (e.g., efflux) by spectrophotometry, flow cytometry, and microscopy.
FAQs
The probe links an optical change to a local biological condition. Binding, enzymatic activity, ion concentration, or membrane environment can alter the fluorescence signal, so the measured light reports where or when that condition occurs. This makes the readout useful not merely for locating a labeled structure, but also for comparing biological states through changes in signal.
Attachment strategy connects the fluorescent signal to a particular biological target or process. Antibody-linked probes can identify structures recognized by those antibodies, nucleic-acid-linked probes can report selected sequences, and small-molecule sensors can respond to local conditions. Choosing among these formats therefore determines whether the experiment emphasizes organization, gene-related information, or dynamic cellular chemistry.
Multiplexed labeling allows researchers to examine more than one labeled target or biological feature in the same investigation. By combining probes that provide distinguishable fluorescence signals, experiments can relate cell organization to molecular or process-level information. This broader view is valuable when interpreting how structures, gene expression, signaling, metabolism, or disease-related changes occur together.
Selection should begin with the biological event or structure that must be measured. Researchers can then match the probe format to the target, such as an antibody, nucleic acid sequence, or small-molecule sensor, and choose an imaging approach capable of detecting its signal. The expected output may be localization, fluorescence change, or comparison of biological conditions.
Fluorescence microscopy can reveal the distribution of labeled biological structures and show changes associated with binding, enzymatic activity, ion concentration, or membrane environment. Depending on the probe, the resulting images or measurements can address cell organization, gene expression, signaling, or metabolism. The outcome is an optical readout that connects observed fluorescence with a defined biological question.
They are useful when researchers need to connect molecular events with their location or cellular context. Applications described for these probes include studying cell organization, gene expression, signaling, metabolism, and disease mechanisms. Their ability to support multiplexed labeling also helps expand biological analysis beyond a single structure or process, while improved probe design can increase sensitivity and specificity.