Photostable dyes suitable for microscopy
Perfect match for the CFP filter set
Highly water soluble and pH-insensitive
Suitable for flow cytometry in the AmCyan channel
CF®405L Dye
A 405 nm-excitable dye with green fluorescence emission
Minimally charged, for less non-specific binding than Alexa Fluor® 488
Narrower emission spectrum for less bleed to red
Very photostable
Compatible with super-resolution imaging by TIRF
Highly water soluble and pH-insensitive
CF®503R Dye
A spectrally unique green dye ideal for spectral flow cytometry
Abs/Em Maxima: 562/584 nm
Extinction coefficient: 100,000
Molecular weight: ~ 714
Excitation laser line: 532 nm or 568 nm
Direct replacement for: Alexa Fluor® 568,
ATTO 565, Rhodamine Red
Features
Yields much brighter antibody conjugates than Alexa
Fluor® 568
Extremely photostable
Excellent choice for multi-color imaging with CF®488A and CF®640R
Compatible with TIRF and multicolor STORM super resolution imaging
Spectrally similar red fluorescent dyes with exceptional brightness
Serves as an alternative for Cy®3.5 and Texas Red®
Rhodamine-based CF®583R can be used as an excellent energy acceptor for FRET when paired with R-PE
Yields the brightest antibody conjugates among spectrally
similar dyes.
Excellent choice for multicolor imaging with green dyes like CF®488A
Extremely photostable
Also see CF®594ST, a version of CF®594 engineered specifically for STORM microcopy
Yields the brightest antibody conjugates among spectrally similar dyes
Far more photostable than Alexa Fluor® 647
Compatible with TIRF, FIONA, and gSHRImP super-resolution imaging methods
CF®680 Features
The brightest among spectrally similar dyes
Validated in multicolor STORM and 3D super-resolution
microscopy
Recommended for LI-COR® Odyssey®
CF®680R Features
Unrivaled photostability among spectrally similar dyes
Compatible with STED and single molecule spectroscopy super-resolution imaging (see pp. 20-21)
Molecular weight compatible with nucleic acid labeling
Recommended for LI-COR® Odyssey®
Brighter near-IR alternative for Alexa Fluor® 700
Superior signal-to-noise for antibodies and other bioconjugates
Exceptional performance for microscopy and flow cytometry
CF®710 Dye
Near-infrared CF® Dye with exceptional photostability
Exceptionally bright and stable
Ideal for in vivo imaging
Recommended for LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
CF Dyes validated in SIM, STORM, STED, TIRF, and more
Superior brightness, photostability, and photochemical switching
Single label antibody conjugates for STORM
Growing list of publications
Figure 1. Normalized absorbance and emission spectra of CF®700 goat anti-mouse conjugate in PBS.
Near-IR CF®700 is one of Biotium’s next-generation long wavelength dyes developed for superior brightness and signal-to-noise.
Other commercial near-IR dyes suffer from problems of limited fluorescence brightness due to excessive dye aggregation and poor stability. As a result of novel molecular engineering by scientists at Biotium, near-IR CF® Dyes overcome these problems, resulting in exceptionally bright dyes for life science research.
CF®700 serves as a superior alternative to Alexa Fluor® 700 for microscopy or flow cytometry. The dye has been validated extensively and offers a brighter signal and higher stain index than Alexa Fluor® 700 for flow cytometry (Figures 2-3).
Superior signal-to-noise for antibodies and other bioconjugates
Exceptional performance for microscopy and flow cytometry
Superior Brightness and Signal-to-Noise Over Alexa Fluor®700
Figure 2. PBMCs stained side by side with antibodies for the same target and of the same clone, conjugated either to CF®700 or to Alexa Fluor® 700. Cells were analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer, and the mean fluorescence of the positive population is plotted. Clones: CD3(SK7); CD4(SK3); CD8(SK1); CD16(3G8); CD19(SJ25C1).
Figure 3. PBMCs stained side by side with antibodies for the same target and of the same clone, conjugated either to CF®700 or to Alexa Fluor® 700. Cells were analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer, and the Stain INdex of the positive population is plotted. Clones: CD3(SK7); CD4(SK3); CD8(SK1); CD16(3G8); CD19(SJ25C1). Stain Index equation: (MFIpos – MFIneg) / 2 x SDneg.
Figure 4. PBMCs stained with CD3(SK7)-CF®700 and CD3(SK7)-Alexa Fluor® 700 (BioLegend® 344821) and analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer.
Figure 5. PBMCs stained with CD4(SK3)-CF®700 and CD4(SK3)-Alexa Fluor® 700 (BioLegend® 344621) and analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer.
Figure 6. PBMCs stained with CD8(SK1)-CF®700 and CD8(SK1)-Alexa Fluor® 700 (BioLegend® 344723) and analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer.
Figure 7. CD3- PBMCs stained with CD16(3G8)-CF®700 and CD16(3G8)-Alexa Fluor® 700 (BioLegend® 302007) and analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer.
Figure 8. PBMCs stained with CD19(SJ25C1)-CF®700 and CD19(SJ25C1)-Alexa Fluor® 700 (BioLegend® 363033) and analyzed in the R712 (APC-700) channel of a CytoFLEX® flow cytometer.
CF is a registered trademark of Biotium, Inc.; Alexa Fluor is a registered trademark of Thermo Fisher Scientific; BioLegend is a registered trademark of Biolegend, Inc; CytoFLEX is a registered trademark of Beckman Coulter, Inc.
Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples.
One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.
See below for our recommended alternatives to Akoya tyramides. Unless noted as a direct replacement, these tyramide dyes are spectrally similar alternatives. They have not been validated as direct replacements for use with Akoya kits or imaging systems.
Note: The CF® Dyes listed here are spectrally similar alternatives to Opal™ dyes, they have not been validated as direct replacements for use with Akoya's kits or imaging system. Opal is a trademark of Akoya Biosciences.
Dyes that carry multiple negative charges can introduce background. Usually, this is more of a concern with labeled antibodies that carry many dyes, as opposed to a small toxin like bungarotoxin. When staining tissues, the endogenous autofluorescence of the tissue itself is often the most significant source of background. Endogenous fluorescence background in tissue is usually highest in the blue wavelengths (DAPI channel) and lowest in the far-red (Cy®5 channel). Our CF®633 bungarotoxin (catalog no. 00009) is a far-red conjugate for the Cy®5 channel with a low negative charge that should have low background from either the dye or autofluorescence.
We test fluorescent bungarotoxin on rat skeletal muscle sections. While the tissue shows autofluorescence, the bungarotoxin staining of motor endplates is usually much brighter than the background for all of the dye colors we’ve tested. However, if you are staining human tissue (especially brain), lipofuscin autofluorescence may be bright in all channels. This usually shows up as bright, punctate dots around cell nuclei. While we would usually recommend our TrueBlack® lipofuscin quenchers for human brain tissue, they are not compatible with bungarotoxin staining. We have, however, found that EverBrite TrueBlack® Mounting Medium (cat. no. 23017) can be used to mount skeletal muscle sections stained with bungarotoxin.
Cy Dye is a registered trademark of Cytiva.
