Fluorescent Product Measurement
Fluorescent Product Measurement - Related Videos
14:39
Measuring Phagosome pH by Ratiometric Fluorescence Microscopy
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Cited by 26 •
2015
Phagosomal pH influences phagosome maturation, oxidant production, phagosomal killing as well as antigen presentation. Here we describe a ratiometric method for measuring time-course and endpoint pH changes in individual phagosomes in living phagocytes using fluorescence microscopy.
07:03
In Situ Measurement of Vacuum Window Birefringence using 25Mg+ Fluorescence
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2020
Presented here is a method to measure the birefringence of vacuum windows by maximizing the fluorescence counts emitted by Doppler cooled 25Mg+ ions in an ion trap. The birefringence of vacuum windows will change the polarization states of the laser, which can be compensated by changing the azimuthal angles of external wave plates.
06:47
Measurement and Analysis of Extracellular Acid Production to Determine Glycolytic Rate
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Cited by 103 •
2015
Glycolysis is a defining metabolic marker in multiple biological systems. Monitoring glycolysis by measuring the extracellular flux of H+ is common, but requires correction to be quantitative and unambiguous. Here, we demonstrate how to gather and correct extracellular flux data to distinguish between respiratory and glycolytic sources of extracellular acidification.
07:00
Measuring Diffusion Coefficients via Two-photon Fluorescence Recovery After Photobleaching
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Cited by 4 •
2010
In this article we will describe the procedure for measuring diffusion coefficients using multi-photon fluorescence recovery after photobleaching. We will begin by aligning the laser along the optical path to the sample and determining the proper experimental parameters, then continue generating and finally fitting fluorescence recovery curves.
10:01
Measurement of Carbon Dioxide Production from Radiolabeled Substrates in Drosophila melanogaster
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Cited by 7 •
2016
This paper describes a method for the measurement of fuel oxidation in Drosophila melanogaster in which trace amounts of specific radiolabeled metabolic substrates are fed to flies. The exhaled radiolabeled CO2 that is a produced from fuel oxidation is collected and measured.
FAQs
The instrument first excites the reaction product at a selected wavelength and then records the light emitted. The recorded intensity serves as a quantitative readout because it can indicate how much product is present when assay conditions are controlled. This optical sequence links the biological reaction to a measurable signal that can be compared across samples or time points.
Controlled conditions make fluorescence comparisons interpretable. When samples are measured under the same conditions, differences in recorded intensity can be related to differences in product concentration rather than changes in the measurement setup. Maintaining this consistency is important for comparing biological samples, reaction rates, or cellular responses within the same experiment or study.
Recording fluorescence at multiple time points shows how the amount of fluorescent product changes during a reaction. The resulting time-dependent signal can be used to compare reaction rates, because faster product accumulation produces a different pattern of change than slower accumulation. This approach allows biological activity to be followed dynamically rather than assessed only at one endpoint.
A typical workflow begins with a reaction that produces a fluorescent product from a nonfluorescent substrate. The product is then measured using a fluorometer or microplate reader configured with a selected excitation wavelength. Researchers record the emitted light under controlled conditions and use the intensity, or its change over time, to assess product formation and biological activity.
Researchers can apply this approach when they need sensitive measurements of enzyme kinetics, metabolic reactions, cellular signaling, or reporter-gene activity. The same measurement strategy connects product formation with different biological questions, allowing investigators to compare reaction rates or responses among experimental conditions. Its value is greatest when small changes in biological activity need to be detected.
The method can provide quantitative fluorescence values, changes in signal over time, and comparisons between reaction rates or biological responses. These outcomes help researchers determine whether activity differs among samples or conditions and can support development of sensitive analytical assays. Interpretation depends on relating the recorded light intensity to product concentration under the controlled conditions of the experiment.