Cyanine 5.18 (or Cy5) is a fluorochrome emitting in the long-red/far-red range, usually regarded as unsuitable for direct observation by the human eye.The 75-nanometer emission window is combined with a wide 60-nanometer excitation passband (590 to 650 nanometers) to allow selective excitation and detection of specific popular .The Cy5 HYQ set employs a bandpass emission (barrier) filter (pass band region between 663 and 738 nanometers) instead of a longpass variety, in order to limit interference from fluorophores emitting outside the intended wavelength region, and to reduce autofluorescence.The 75-nanometer emission window is combined with a wide 60-nanometer excitation passband .

Cy5color

Frick Chemistry Laboratory Princeton University Corner of Washington Road and Scholar Way Princeton, NJ 08544 Front office: 609-258-3900

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Cy5vs Alexa 647

The Ultrafast Laser Spectroscopy facility in the Frick Chemistry Laboratory offers state-of-the-art equipment for measuring dynamics in photoactive materials on femtosecond-to-millisecond timescales. The facility in Frick Chemistry Laboratory is open to members of any department at Princeton University. Users from outside the University are welcome, as well. The manager of the facility performs the measurements, as well as offers training for long-term users.

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About PE-Cy5. PE-Cyanine®5 (PE-Cy5) is a red-emitting tandem fluorophore that combines phycoerythrin (PE) and Cy5. The donor molecule, PE can be excited by the 488-nm blue, 532-nm green, or 561-nm yellow-green laser and and transfers energy to the acceptor molecule, Cy5. PE-Cy5 has an excitation peak at 496 nm and an emission peak at 667 nm.Human blood was blocked with normal goat serum and incubated with an anti-CD3 mouse monoclonal antibody; cells were washed, resuspended and incubated with either an Alexa Fluor 647 or Cy5 goat anti–mouse IgG secondary antibody. Samples were analyzed on a flow cytometer equipped with a 633 nm He–Ne laser and a longpass emission filter (>650 nm).Cy2, Cy3 and Cy5 Conjugates for Permanent Mounting. . Approximate peak wavelengths of excitation and emission for Cyanine fluorophores conjugated to affinity-purified antibodies. Only approximate values are given for purposes of comparing one fluorophore with another. Actual values may vary depending on the spectrofluorometer used in each .Cy5.5 (Cyanine-5.5) is a fluorescent compound with an excitation peak at 683 nm and an emission peak at 703 nm. Other spectra of interest include: Cy5.5NS, Cy2, and iFluor 680.Cy5.5 (Cyanine-5.5) belongs to the following categories: Amine Reactive Dyes and Probes for Conjugation, and Cyanines. Purchase Cy5.5 (Cyanine-5.5).

Cy7excitation emission

The facility allows measurements of time-resolved change of the absorption spectra (Transient Absorption) with femtosecond (10-14 s) time resolution and up to microsecond timescales with spectral windows in UV-VIS-NIR, allowing probing all possible excited state transitions. It consists of LIBRA Ti:Sapphire amplifier (Coherent Inc.), OPerA Solo optical parametric amplifier (Coherent Inc.), Helios transient absorption spectrometer (Ultrafast Systems), and EOS transient absorption spectrometer (Ultrafast Systems). The Ti:Sapphire amplifier delivers 45fs pulses at 1kHz rate, with total power of 4.0W at 800nm wavelength. Optical parametric amplifier allows tuning of the excitation wavelength in the spectral range from 290 – 2600nm, with average power of 50mW and pulse duration 50fs. Transient absorption spectrometer allows measurements in 350 – 1600nm spectral region (probing range), and on <1ms time range.  A recent addition is the MID-IR system (Ultrafast Systems) for direct probing vibrational modes in molecules (3500cm-1 to 800cm-1).  Allowing for the direct monitoring of molecular bonds on light-initiated processes.  This can even be extended to millisecond timescales for monitoring of diffusion limited processes.

Cy5absorbance

The fluorophore has its emission maximum in the red region, where many CCD detectors exhibit maximum sensitivity, and biological objects show low background. The dye color is very intense, therefore quantities as small as 1 nmol can be detected in gel electrophoresis by naked eye. . Cy5 excitation and emission spectra. Customers also .Supplementary Methods: TIR excitation and emission set up. NIHMS495771-supplement-Supplementary_materials.doc (102K) . 40% of Cy5 emission leaks into the Cy5.5 channel if Cy5.5 is used as a second acceptor), but careful calibration and correction can recover the original intensities 43. Our recent test indicates that Cy7 is a promising .

Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength.

Minimal excitation by red diode lasers, very low background, good stability, and consistent emission . laser power greater than 150 mW. US Orders: 877.232.8995 bdbiosciences.com Features PerCP PerCP-Cy5.5 Absorption max/emission max 490 nm/677 nm 490 nm/695 nm Excitable by 488-nm laser Yes Yes Spillover to PE Channel Minimal Minimal Relative .Peridinin chlorophyll protein-Cyanine5.5 (PerCP-Cyanine5.5) is a photostable tandem dye excited from the 488 nm laser. It is stable in fixative, and stained cells can be stored overnight.

(Cy5 ® ) R e l a t i v e I n t e n s i t y 350 400 500 550 600 650 700 750 800 450 Wavelength (nanometers) Excitation Emission PE-Cy 5® Laser line (488) Tandem (PE-Cy5 ® ) Excitation and Emission Spectral Profiles Energy Level Diagrams Fluorochromes have unique and characteristic spectra for absorption (excitation) and emission. A single dyeBroad UV Excitation 290-365 nm Emission Filters LP = Long pass filter DF = Band pass filter SP = Short pass filter 555 nm 555LP Molecular Imager FX Filters VersaDoc Filters 520 nm 520LP 660 nm 515 nm . Poor _ Cy5: 650 nm ____ Poor ___ Cy5.5: 675 nm ____ Emission Maxima ____ 443 nm: Coumarin ____ 461 nm: DAPI ____ 506 nm: Cy2Excitation maximum: λ exc = 649 nm Emission maximum: λ em = 670 nm Extinction coefficient: ε max = 250,000 mol-1 cm-1 Correction factor: CF 280 nm = 0.05. Description: Fluorescence techniques have become a major tool in biological sciences. Cy5.5 has strong two-photon absorbance at 1.25 µm and emission that peaks at 710 nm. In the approach, Cy5.5–dextran is delivered by retro-orbital injection and spreads into all vessels in the body.

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Cy5spectrum

Cy5 is available conjugated to: Secondary Antibodies for Multiple Labeling; Streptavidin; ChromPure™ Purified Proteins from Normal Serums (Donkey and Goat) Cy2, Cy3 and Cy5 Conjugates for Permanent Mounting. Excitation Peak (nm) Emission Peak (nm) . Show Excitation and Emission Spectra . The primary photodegradative mechanism of cyanine dyes is dependent on singlet oxygen generated by energy transfer from excited triplet fluorophores to molecular oxygen 24,31 ().Singlet oxygen .588–640-nm excitation filter range, distinctly below the excitation optimum for Cy5, (c) a 650–700-nm emission filter range, transmitting the low-wavelength portion of Cy5 emis-sion, and (d) high-efficiency filter set components allowing a narrow gap between excita-tion and emission ranges without visible cross-talk of excitation light in .PE-Cyanine®5.5 (PE-Cy5.5) is a red-emitting tandem fluorophore that combines phycoerythrin (PE) and Cy5.5. The donor molecule, PE can be excited by the 488-nm blue, 532-nm green, or 561-nm yellow-green laser and and transfers energy to the acceptor molecule, Cy5.5. PE-Cy5.5 has an excitation peak at 496 nm and an emission peak at 695 nm.

Invitrogen Cyanine5 (Cy5) dye is a bright, far-red-fluorescent dye with excitation designed for use with the 633 nm or 647 nm laser lines. A significant advantage to using long wavelength dyes such as Cy5 dye or Invitrogen Alexa Fluor 647 dye over other fluorophores is the low autofluorescence of biological specimens in this region of the spectrum.

Texas Redexcitation emission

Fluorescence emission spectra of R-Phycoerythrin and PE-Cy5. Excitation is at 488 nm and the emission spectra are normalized to the absorbance of both dyes at 488 nm. PE-Cy5 has a peak emission spectrum at 669 nm where there is little overlap with the R-PE spectrum. This reduces compensation required to less than 10%. Excitation filters are of 35-nm bandwidth centered at the wavelength indicated and emission filters DsRed and Cy5.5 are 575–650 and 695–770 nm, respectively. Pseudocolor scale bar: radiant .In the second experiment, intensities of Cy3 and Cy5 were measured alternatively, switching from Cy3 excitation/emission condition to Cy5 using the same sam- ple. Spectroscopic measurements were .

Cy5Structure

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Several options for sample mounting are available in order to prevent photo-bleaching/degradation: stirring, translation, and purging with dry N2.  Helios/EOS software includes dispersion compensation algorithm, anisotropy calculation and other useful tools. The measurements at different excitation intensities, wavelengths or at different temperatures of the sample can be automated. Global analysis of the data (three-dimensional array of time-resolved spectra) allows to map the exact pathways and timescales of the processes occurring in the materials of interest. This software is free and can be downloaded from the internet together with the manual and instructions.

Cy3excitation emission

Standard excitation and emission spectra of Cy5 conjugate are provided for comparison (“Cy5 exc” and “Cy5 em,” respectively). Note rapidly increasing efficiency when excitation extends above 620 nm, parallel to ascending slope .

Cyanine5 (Cy5) is a bright, far-red-fluorescent dye with excitation suited for the 633 nm or 647 nm laser lines and is used for imaging, flow cytometry and genomics. See moreProtocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, . See moreA handy reference poster featuring the broad range of our available dyes and labeling reagents. Request the guide See moreA tool for selecting the optimal fluorescent dyes for your Invitrogen EVOS cell imaging systems. Download the guide See more

Our broadband excitation comfortably covers absorption of both the Cy3 and Cy5 (Fig. 3b), and, in contrast to standard donor-lifetime FLIM, we detect emission from the acceptor Cy5 only. Notably, to construct our far-red HVI we used Cy5 as the fluorophore because of its desired excitation/emission spectrum (ex. peak at 649 nm; em. peak at 670 nm) and high molecular brightness .

However, these inner filter effects are avoided when the Cy5 emission spectra for 1T or 2T are generated with 350 nm where there is much less absorption of the ultraviolet excitation light by other Cy5-cassette molecules in the solution.Crucial factors were (a) an excitation path in the microscope not absorbing light in the orange-red range, up to 640 nm, (b) a 588-640-nm excitation filter range, distinctly below the excitation optimum for Cy5, (c) a 650-700-nm emission filter range, transmitting the low-wavelength portion of Cy5 emission, and (d) high-efficiency filter set .By modulated excitation of a long wavelength Cy5 transient absorption, we dynamically modulate Cy5 emission. The frequency dependence enables specificaton of the dark state timescales enabling optical demodulation-based signal recovery from high background. These dual laser illumination schemes for high sensitivity fluorescence signal recovery .CFP excitation coupled with a YFP emission filter. For FRET sensitized-emission donor-acceptor bleedthrough measurements. Requires CFP and YFP light cubes for CFP-YFP FRET experiments. AO (470/22 nm Excitation; 488LP nm Emission) Cat. No. AMEP4970

Ultrafast laser spectroscopy is a standard method in a number of scientific fields (physics, chemistry, molecular biology, engineering etc.) and is used both as a primary research technique, as well as an accessory characterization tool for supporting the interpretation of the experiments. This technique provides detailed information on the dynamic processes in photoactive materials (in solid and liquid phase). These processes can be excited state relaxation, both inter and intramolecular energy/electron transfer, conformational change (e.g. isomerization), chemical reaction dynamics.

Excitation and emission. Cy5 has an excitation peak at 651 nm and an emission peak at 670 nm. It is well excited by the 633 nm or 647 nm laser lines. For detection, a .Cy5.5 is an important historical fluorophore from GE Healthcare. It has an excitation peak at 678 nm and an emission peak at 695 nm. Spectrally its similar to Alexa Fluor™ 680.PE-Cyanine®5 (PE-Cy5) is a red-emitting tandem fluorophore that combines phycoerythrin (PE) and Cy5. The donor molecule, PE can be excited by the 488-nm blue, 532-nm green, or 561 .PE-CyTM5 (Ex max496 nm/Em. max667 nm) is a tandem conjugate that combines phycoerythrin and the cyanine dye Cy5. Because of its broad absorption range and the fact that its emission .