What is the wavelength of FITC?
What is the wavelength of FITC?
475 to 650 nm
FITC emits fluorescence from 475 to 650 nm, peaking at 525 nm, which falls in the green spectrum.
What is the color of FITC?
FITC exhibits an excitation maximum at λ = 495 nm and emission maximum at approximately λ = 519 nm. The color of the compound is yellow while the emitted light is green.
How do you choose excitation wavelength for fluorescence?
the range of excitation wavelength should be 200 nm to 20 nm less than your emission wavelength. 6. Now the last point, check the excitation spectra and find out the highest intense peak. The wavelength belongs to highest intense peak, is the suitable excitation wavelength for ur sample.
What wavelengths cause fluorescence?
In general, fluorescence investigations are conducted with radiation having wavelengths ranging from the ultraviolet to the visible regions of the electromagnetic spectrum (250 to 700 nanometers).
What is FITC A in flow cytometry?
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC has excitation and emission spectrum peak wavelengths of approximately 495 nm and 519 nm, giving it a green color. Like most fluorochromes, it is prone to photobleaching.
What is FITC flow cytometry?
Why is fluorescence red shifted?
Red shifts occur because long-wavelength excitation results in photoselection of those fluorophores which are interacting most strongly with the polar solvent molecules.
How is the excitation wavelength prevented from reaching your eyes in a fluorescence microscope?
The excitation wavelength, such as blue, is focused by the objective lens on the sample, which prevents it from reaching your eyes. The wavelengths that pass through provide the colored light that emerges from the objective lens.
Why is fluorescence red shift?
Why is fluorescence red?
As the solution strength is decreased, three-dimensional fluorescence maxima systematically shifted to shorter wavelengths. They describe the effect as ‘cascade’ effect, thereby implying excited state energy transfer to be the primary cause of the red-shift.
Is Cy3 red or green?
Cy3 fluoresces greenish yellow (~550 nm excitation, ~570 nm emission), while Cy5 is fluorescent in the far-red region (~650 excitation, 670 nm emission). Cy3 can be detected by various fluorometers, imagers, and microscopes with standard filters for Tetramethylrhodamine (TRITC).
What is the wavelength of a flow cytometer?
The most common lasers used in traditional flow cytometers are 488 nm (blue), 405nm (violet), 532nm (green), 552nm (green), 561 nm (green-yellow), 640 nm (red) and 355 nm (ultraviolet). Additional laser wavelengths are available for specialized applications.
What is FITC fluorescence color?
Fluorescence color usually refers to the color of light a fluorophore emits at its highest stable excited state. However, if FITC fluorescence is detected only via channel B (Figure 9), it will appear orange and be much weaker in intensity.
Which fluorescent dye should I use for flow cytometry?
Applications where a smaller dye is required are more appropriate for these dyes. These fluorochromes are well suited for fluorescent microscopy. Fluorescein isothiocyanate (FITC) is currently the most commonly used fluorescent dye for flow cytometry analysis.
What does the FITC spectrum look like?
However, FITC’s emission is not restricted to green photons; it also emits yellow, orange, red, albeit at lower probabilities. You may notice something curious about the spectrum if you look closely at the emission curve immediately around the excitation line.