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
Compatible with 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
Compatible with LI-COR® Odyssey®
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
CF®740 Dye
Near-infrared CF® Dye with exceptional photostability
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
APC-Astral™813
Novel long Stokes shift tandem dye for flow cytometry multiplexing
Abs/Em Maxima: 797/816 nm
Extinction coefficient: 210,000
Molecular weight: ~3334
Excitation laser line: 785 nm
Spectrally similar to: Indocyanine Green
Features
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Dye options for STORM & photoacoustic imaging
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Optimal for the 808 nm laser for flow cytometry
Exceptionally bright and stable
Ideal for in vivo imaging
Compatible with LI-COR® Odyssey®
Superior signal-to-noise for bioconjugates
Optimal for the 808 nm laser for flow cytometry
CF® Dyes for Multi-Color Super-Resolution Microscopy
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 absorption and emission spectra of CF®568 goat anti-mouse antibody conjugate in PBS.
CF®568 is a red fluorescent dye with an excitation spectrum optimally matching the 568 nm laser line. Antibody conjugates of the dye are much brighter than those of Alexa Fluor® 568. CF®568 is also more photostable than Alexa Fluor® 568, making CF®568 a much better choice for demanding applications, such as confocal and single molecule imaging. CF®568 has been validated in super-resolution imaging by SIM, TIRF, STED, and multicolor STORM, and produces better STORM images than Cy®3b (learn more about CF® Dyes for Super-Resolution Imaging). See CF®568 product lines below.
CF®568 Technical Summary
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
Figure 2. CF®568 produces brighter antibody conjugates than Alexa Fluor® 568. Jurkat cells were stained with mouse anti-CD3 or isotype control, followed by CF®568 or Alexa Fluor® 568 goat anti-mouse conjugates with varying degrees of labeling (DOL, dye molecules per protein molecule). Fluorescence was analyzed on a BD FACSCalibur™ flow cytometer in the FL2 channel; bars represent geometric mean fluorescence.
Figure 3. CF®568 is more photostable than Alexa Fluor® 568. Jurkat cells were stained with biotinylated anti-CD3 antibody followed by CF®568 or Alexa Fluor®568 streptavidin conjugate. Cells were continuously exposed to mercury arc lamp excitation through a Cy®3 filter cube. Images were captured every 15 seconds for 5 minutes, and fluorescence intensity was normalized to time zero.
Figure 4. MCF-7 cells stained with CF®568 monoclonal anti-Ep-CAM (EGP40/826) (red) and Hoechst (blue).
Figure 5. HeLa cells stained with NucSpot® 470 (green) and CF®568 phalloidin (red).
Figure 6. Section of rat intestine stained with CF®568 phallodin (red). Nuclei are stained with DAPI (blue).
CF is a registered trademark of Biotium, Inc; Alexa Fluor is a registered trademark of Thermo Fisher Scientific.
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.
