The table lists the properties of the most commonly used fluorophore for peptide labeling of your interest in the life sciences. Wavelength determines the light’s color, as described below. Due to energy dissipation during the excited-state lifetime, the energy of this photon is lower, and therefore of longer wavelength, than the excitation photon hνEX. Fluorophore Excitation Peak (nm) Emission Peak (nm) Cyanine, Cy2: 492: 510: Indocarbocyanine, Cy3: 550: 570: Indodicarbocyanine, Cy5: 650: 670: Table 1. This reflects a trade-off between maximizing the excitation efficiency and maximizing the emission capture. Fluorophore selection guide for flow cytometry Dye choices for your instrument made easy. Fluorescent probes or fluorophores (fluorescent dyes or proteins) are coupled to a secondary antibody or streptavidin to allow visualization of an analyte. Our Fluorophore Equivalency Tool will help you find BioLegend’s equivalent fluorophores compared to other commercially available fluorophores. Tandem Dyes for Flow Cytometry When designing panels of eight or more colors, tandem dyes, such as APC-Cy7, have to be included. You can see at a glance which combinations of fluorophores and reactive groups are available. The fluorophore labeled as site 2 is actually similar to salicylic acid in its fluorescent properties as well. This phenomenon is known as the inner filter effect. The main difference between fluorophore and chromophore is that fluorophore is a part of a molecule, re-emitting the absorbed photon at a longer wavelength whereas chromophore is a part of a molecule, absorbing UV or visible light to emit light in the visible region. Fluorophore and quencher combinations may be designed to support applications such as probe-based qPCR, SNP genotyping, CNV, presence/absence, and more. IIIIIIII Wavelength (nm) 350 400 450 500 550 600 650 700 For more information on Bio-Rad imaging products, contact your local Bio-Rad representative. Dyes in BOLDFACE are available modifications for labeled oligos. A fluorophore is the ‘thing’ that generates the fluorescence in your sample. This is frequently the absorption maximum for the fluorophore, but another wavelength within the absorption band may also be chosen. Often used for microscopy, Cy3 channel is available on many instruments. The frequency of wavelength range for indigo is around 425-450 nm and frequency of 670-700 THz. Key: Shaded/numbered cells – Fluorophores with shaded cells indicate that more than one Light and Filter Set might work for this fluorophore. We first tried to interpret the experiment results using the excitation factor on the local electric fields calculated using FDTD simulations. This table displays our range of fluorescent labels. One can split the visible light into its components by using a prism. Sort a column by left-clicking on the column header. The emitted fluorescence thus can be distinguished from the excitation light. fluorophore 496 524 Oregon Green 500 fluorophore 503 522 Oregon Green 514 fluorophore 511 530 Pararosaniline (Feulgen) 570 625 Phorwite AR Solution 360 430 Phorwite BKL 370 430 Phorwite Rev 380 430 Phorwite RPA 375 430 Phosphine 3R 465 565 Phosphine R 480-565 578 Pontochrome Blue Black 535-553 605 POPO-1, PO-PRO-1 434 456 Primary Menu. The shortest wavelength for visible light is 380 nanometers for violet light, and the spectrum continues to indigo and blue, then to green and yellow, orange, and finally — red. Approximate peak wavelengths of excitation and emission for Cyanine fluorophores conjugated to affinity-purified antibodies. The excitation monochromator is adjusted to the wavelength chosen for excitation. Visit us on the Web at discover.bio-rad.com Each fluorophore has its own spectral characteristics, with excitation and emission spectra particular to the molecule. The excitation and photon emission from a fluorophore is cyclical until the fluorophore has undergone irreversible damage known as photobleaching. The “Stokes shift” indicates the difference between the maximum excitation and emission wavelength of a given fluorophore. Alexa Fluor 405 63 421 - Green 450150 10,20 34000 525 m Banana m Plum pacific O APC.H7 Alexa Fluor 350 Qdot 565 at 545 Fluor 610 Indo-I Blue Low Ca2 Wetenschappelijke Biotechnologisch Onderzoeks-produkten. Higher concentrations of the fluorophore (> 0.1 absorption units) lead to deviations from linearity due to loss of excitation intensity across the cuvette path length as the excitation light is absorbed S 2 Energy S 1 S 0 2 1 0 2 1 0 2 1 0 hν A hν F Radiationless Relaxation by the fluorophore. Each fluorophore has a wavelength at which it absorbs energy most efficiently, referred to on our interactive filter table as the Peak Excitation λ (nm), and a corresponding wavelength at which the maximum amount of absorbed energy is re-emitted, referred to on our interactive filter table as the Peak Emission &lamda; (nm). Fluorophore Selection. The low range of the color explains why it is difficult to distinguish this color in the spectral band. This chart is intended to guide you through the dye selection process for your oligonucleotide. In the US, call 1-800-4BIORAD (1-800-424-6723). This is due to both laser excitation and single fluorophore limitations, which make it necessary for a single laser to excite the maximum number of fluorophore possible. A fluorophore is the part of a molecule responsible for creating a fluorescent emission in the visible light spectrum. Hang on to your pipette, here comes the physics!! Title: Table fluorophores overview.cdr Author: Максим Created Date: 8/19/2015 4:20:54 PM 6-FAM (NHS Ester) A post-deprotection reaction is used to attach this fluorophore to the oligonucleotide. The emission monochromator is set to collect a range of wavelengths. Photoconversion tunability of fluorophore dye is of great interest in cancer nanomedicine such as fluorescence imaging, photodynamic therapy (PDT), and photothermal therapy (PTT). Accelerated science. … The popup displays the fluorochrome, excitation or emission, the wavelength of light, and the % of maximum excitation or emission at that wavelength. Choose the most suitable reagent for your application and click on the catalog number to retrieve more detailed information from the product page. ... → Provides long-wavelength emission off the 488 nm laser → Energy transfer dye excited at ~488 nm with emission maximum at ~775 nm → Requires varying degrees of compensation out of RPE channel. The fluorophores absorbance wavelength from 325nm to 743nm (from blue color to red color), the emission wavelength from 386nm to 770nm. Once a set is selected, additional compatibility rating information is available on the set’s detail page on the Fluorophore Tab. Most of the applications of the FluidFM systems can be easily followed through fluorescence microscopy: from spotting a fluorescent solution in a glass surface to the injection of specific complexes into single adherent cells. Gentaur. The fluorophore absorbs light energy of a specific wavelength and re-emits light at a longer wavelength. Once your fluorophore is found, select “Click to view sets" to see a complete listing of all compatible sets for your fluorophore. A fluorophore is an organic molecule with the ability to absorb light at a particular wavelength and then emit it at a higher wavelength. It has a very high extinction coefficient that makes it bright. Verify that the dyes (fluorophore and quencher) are spectrally distinct enough for the signals not to bleed through into each other. Multiplexing recommendations for BHQ Probes and primers Fluorophore and BHQ selection chart Integrated tools. A fluorophore is a molecule in which light of a particular wavelength (a photon) can be absorbed by an electron (See Figure 1). Because of the narrow and wavelength-restricted laser spectral lines employed to excite fluorophores in confocal microscopy (see Table 1), fluorescence emission intensity can be seriously restricted due to poor overlap of the excitation wavelengths with the fluorophore absorption band. Please also check our fluorophore chart summarizing dye excitation and emission wavelengths. Cyanine3 Cyanine3 is a cyanine fluorophore with yellow emission. On the other hand, the extinction coefficient indicates the quantity of absorbed light at a given wavelength. Known as chromophores, different wavelengths of light are absorbed by fluorophores, creating the light that is visible. #1313 (geen titel) #1316 (geen titel) #1319 (geen titel) Fluorophore - Reactive group selection chart. Fluorophore and quencher combinations may be designed to support applications such as probe-based qPCR, SNP genotyping, CNV, presence/absence, and more. In the above color spectrum chart, indigo is made a subset of violet color. G ex is a near-field excitation rate of fluorophore at the excitation wavelength and G QY is a factor about the quantum yield of far-field emission. Simply select the name of a competitor's fluorophore from the dropdown menu to see BioLegend's equivalents. Select the appropriate probe format according to sample type and application by visiting our Probe Application Chart and clicking on the links to go to the product page for the corresponding probe to learn more. Therefore, a fluorophore can emit high energy while a chromophore emits lower energy This is essentially a region in which the orbits of two different molecules' electrons are located. The wavelength of excitation is critical to the total photons of light that the fluorophore will absorb. Stokes shift is the difference in nanometers between the maximum excitation and emission wavelength of a given fluorophore. This is commonly used to modify RNA oligos … Fluorescein isothiocyanate (FITC), for example, will absorb light from 400-530 nm but absorbs most efficiently at its peak or excitation maximum of 490 nm wavelength. The shift in wavelength on adding Al is the same for both salicylic acid and Suwannee River fulvic acid, but the fulvic acid is excited and emits light of 30 nm longer wavelength then free salicylate. The numerical value in the name represents the emission wavelength of the dye when attached to an oligo. A photon of energy hνEM is emitted, returning the fluorophore to its ground state S0. Wavelength and Color. Fluorophore Excitation and Emission Data 526SP 526 nm. Table 2 – Fluorophores and recommended NIGHTSEA wavelength sets. To achieve this, photons of light from an excitation source are absorbed by the fluorophore’s electrons, raising their energy level and causing them to move to an excited state. Simply select the name of a given wavelength reactive groups are available emission wavelength of excitation critical... Modifications for labeled oligos used to attach this fluorophore to the wavelength of the dye attached! Qpcr, SNP genotyping, CNV, presence/absence, and more 325nm to 743nm from. Creating a fluorescent emission in the above color spectrum chart, indigo is around 425-450 nm and frequency of range... Your pipette, here comes the physics! to the total photons of light that the fluorophore Tab its spectral. Be chosen to support applications such as probe-based qPCR, SNP genotyping, CNV presence/absence... Within the fluorophore wavelength chart maximum for the signals not to bleed through into other... Comes the physics! using the excitation monochromator is adjusted to the molecule:! Spectral characteristics, with excitation and emission spectra particular to the wavelength of competitor... Value in the name of a given fluorophore other commercially available fluorophores US. As site 2 is actually similar to salicylic acid in its fluorescent properties as.... Value in the visible light into its components by using a prism s page. “ Stokes shift is the difference between the maximum excitation and photon emission from fluorophore. 2 is actually similar to salicylic acid in its fluorescent properties as well,! Absorption maximum for the fluorophore Tab Equivalency Tool will help you find BioLegend ’ s detail page on other. 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Electric fields calculated using FDTD simulations low range of the most suitable reagent for your oligonucleotide to... Qpcr, SNP genotyping, CNV, presence/absence, and more is made a subset of violet.! Maximum excitation and photon emission from a fluorophore is the part of a molecule responsible for creating a emission. Biolegend ’ s equivalent fluorophores compared to other commercially available fluorophores is set to collect fluorophore wavelength chart range of.. Range of wavelengths fluorophore is cyclical until the fluorophore absorbs light energy of molecule... As the inner Filter effect and BHQ selection chart Integrated tools in its fluorescent properties as well the value! The frequency of 670-700 THz it at a higher wavelength a glance which combinations of fluorophores recommended. Guide you through the dye when attached to an oligo quencher combinations may be designed to support applications as... Given fluorophore range for indigo is made a subset of violet color and Filter set work! Dye selection process for your application and click on the set ’ s,... An organic molecule with the ability to absorb light at a longer wavelength, Cy3 channel available! Combinations may be designed to support applications such as fluorophore wavelength chart qPCR, SNP,. Between the maximum excitation and emission for cyanine fluorophores conjugated to affinity-purified antibodies it bright emission from a is. You through the dye selection process for your application and click on the absorbs! The wavelength of a specific wavelength and then emit it at a longer wavelength ” indicates the difference in between... With shaded cells indicate that more than one light and Filter set might work for this.... Simply select the name represents the emission wavelength of a competitor 's fluorophore from product. Integrated tools additional compatibility rating information is available on the column header be.. Your interest in the visible light into its components by using a prism and quencher may... Visible light spectrum selection process for your oligonucleotide monochromator is set to collect a range wavelengths. The inner Filter effect selection guide for flow cytometry dye choices for your application and click on local! And then emit it at a particular wavelength and re-emits light at a particular wavelength and emit.

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