How do you choose fluorophore for flow cytometry?

Selecting the Right Fluorophores for Flow Cytometry Experiments Understand your flow cytometer. ... Consider target abundance. ... Research fluorophore properties. ... Increase panel size with tandem dyes. ... Think about using calibration and compensation beads. ... Never underestimate the importance of compensation controls.

What is the most commonly used fluorescent dye?

isothiocyanate (FITC) is an organic fluorescent dye and probably one of the most commonly used in immunofluorescence and flow cytometry.

How do I choose a fluorophore?

Select fluorophores with high extinction coefficients ( ) One defining factor of a fluorophore's brightness is its extinction coefficient (a measurement of the probability of absorbing a photon of light); the higher the value of the extinction coefficient the brighter the fluorophore.

How do I choose fluorescence dye?

Here the most important aspects to consider when you select a fluorescent dye: The extinction coefficient at the excitation wavelength should be as high as possible. ... The quantum yield should be as high as possible. ... The fluorescent dye should be photostable. ... Photochromic behavior. ... Live/dead cell permeability.

How do I choose the best fluorophore?

Select fluorophores with high extinction coefficients ( ) One defining factor of a fluorophore's brightness is its extinction coefficient (a measurement of the probability of absorbing a photon of light); the higher the value of the extinction coefficient the brighter the fluorophore.

How do you choose fluorophore for flow cytometry?

Selecting the Right Fluorophores for Flow Cytometry Experiments Understand your flow cytometer. ... Consider target abundance. ... Research fluorophore properties. ... Increase panel size with tandem dyes. ... Think about using calibration and compensation beads. ... Never underestimate the importance of compensation controls.

Which fluorescent dye can be used for fluorescence?

Rhodamine. The spirolactam structure of rhodamine is nonfluorescent and colorless and exhibits high selectivity for metal ions; the rhodamine ring in its open form has strong fluorescence.

What information do you need to know while choosing fluorophores to use in your flow cytometry experiments?

When selecting a panel of fluorophores for your flow cytometry panel, it is important to follow a few basic guidelines. First, it is important to understand your flow cytometer. You should know the number and types of lasers present, their excitation capabilities and how to set your filters.

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Fluorophore selection guide for the Attune NxT Acoustic Focusing Cytometer UVIR 300Excitation laser400Emission filter (nm)ChannelRecommended dyes440/50VL1Alexa Fluor 405 Pacific BlueViability dyes.

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