Sony Biotechnology offers over 9,000 high quality monoclonal antibody products to support your research. These include widely published clones conjugated to bright dyes such as Brilliant Violet, and PE/Dazzle.
Browse the technical tools section for practical advice on fluorochromes, panel design, as protocols for surface and intracellular (cytokine, transcription factor, and phosphoprotein) staining. The Biology section maps markers and reagents appropriate to T-cell, B-cell, Cancer and Stem Cell investigations. This is where you’ll also find sample panels.
Fluorochromes
Sony Biotechnologies is committed to advancing science through innovations in both flow cytometry instrumentation and reagents. At Sony we are committed to increasing the number and quality of fluorochromes available to continue to improve your results. Please visit our website and speak with your sales representative to learn about our latest releases.
Over the last several years the number of fluorochromes available for flow cytometry has dramatically increased. These next few pages will provide basic information about our current fluorochrome selection.
When selecting fluorochromes it is important to know the laser and filter configuration of your instrument. Spectral overlap is also a consideration. With compensation one can separate the overlap however some signal will be lost. This is especially true with a conventional flow cytometer compared to a spectral analyzer(more about this in panel design).
For example, two of the most common fluorochromes, FITC (blue) and PE (orange) have significant overlap. With conventional flow cytometers the overlapping signal is subtracted decreasing the overall detection efficiency, With antigens expressed at a high density such as CD4 this is not a problem. However lower density or variable density antigens may be difficult to resolve. It is important to note that the Sony SP6800 and SA3800 Spectral Analyzers use a different mechanism to capture all fluorescence negating these issues.
Fluorochromes for Blue Laser (488 nm)
The Blue (488nm) Laser is the most common laser for flow cytometry research. Blue lasers are generally equipped to detect 2-5 flurochromes.
Emission Spectra of Fluorochromes excited by the Blue Laser
Alexa Fluor® 488
Alexa Fluor® 488
Relative brightness
+++
Recommended filter
530/30
Excitation max (nm)
495
Emission max (nm)
519
Recommended for
Medium to high density markers; intracellular markers
Incompatible dyes
FITC, BD Horizon Brilliant™Blue 515
Comments
FITC
FITC
Relative brightness
+++
Recommended filter
530/30
Excitation max (nm)
493
Emission max (nm)
525
Recommended for
Medium to high density markers
Incompatible dyes
Alexa Fluor®488, BD Horizon Brilliant™Blue 515
Comments
PE
PE
Relative brightness
+++++
Recommended filter
575/26
Excitation max (nm)
565
Emission max (nm)
575
Recommended for
Low density markers
Incompatible dyes
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers
PE/Dazzle™ 594
PE/Dazzle™ 594
Relative brightness
+++++
Recommended filter
575/26
Excitation max (nm)
565
Emission max (nm)
575
Recommended for
Low density markers
Incompatible dyes
PE-CF594
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers
PE-Cy™ 5
PE-Cy™ 5
Relative brightness
+++++
Recommended filter
670/14
Excitation max (nm)
565
Emission max (nm)
670
Recommended for
Low density markers
Incompatible dyes
PerCP, PerCP-Cy™5.5
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers, Tandem dye
PerCP
PerCP
Relative brightness
+++
Recommended filter
695/40
Excitation max (nm)
482
Emission max (nm)
675
Recommended for
Medium to high density markers
Incompatible dyes
PE-Cy5, PerCP-Cy5.5
Comments
PerCP-Cy 5.5
PerCP-Cy 5.5
Relative brightness
++
Recommended filter
695/40
Excitation max (nm)
482
Emission max (nm)
690
Recommended for
Medium to high density markers
Incompatible dyes
PerCP, PE-Cy5
Comments
Tandem dye
PE-Cy™7
PE-Cy™7
Relative brightness
++++
Recommended filter
780/60
Excitation max (nm)
496
Emission max (nm)
785
Recommended for
Low density markers
Incompatible dyes
APC-Cy™7 (on instruments with collinear lasers)
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers; Tandem dye is sensitive to degradation. Protect from light and avoid prolonged exposure to paraformaldehyde containing fixatives.
Fluorochromes for the Red Laser (638 nm)
Red lasers are another popular option for flow cytometry. Flow cytometers for the red laser can typically detect 1-3 fluorochromes.
Emission Spectra of Fluorochromes excited by the Red Laser
APC
APC
Relative brightness
++++
Recommended filter
660/20
Excitation max (nm)
650
Emission max (nm)
660
Recommended for
Medium/Low density markers
Incompatible dyes
Alexa Fluor®647, eFluor®660
Comments
Alexa Fluor®647
Alexa Fluor®647
Relative brightness
++++
Recommended filter
660/20
Excitation max (nm)
650
Emission max (nm)
668
Recommended for
Medium/Low density markers; intracellular markers
Incompatible dyes
APC, eFluor®660
Comments
Alexa Fluor®700
Alexa Fluor®700
Relative brightness
++
Recommended filter
730/45
Excitation max (nm)
696
Emission max (nm)
679
Recommended for
High density markers
Incompatible dyes
BD Horizon™ APC-R700
Comments
APC-Cy™7
APC-Cy™7
Relative brightness
+
Recommended filter
780/60
Excitation max (nm)
650
Emission max (nm)
785
Recommended for
High density markers
Incompatible dyes
APC-H7, PE-Cy™7 (on instruments with collinear lasers)
Comments
Tandem is sensitive to degradation. Protect from light and avoid prolonged exposure to paraformaldehyde containing fixatives
Fluorochromes for Violet Laser (405 nm)
Violet lasers have increased in popularity in recent years due to the availability of many new bright dyes including the Brilliant Violet™ family. Most violet lasers can detect 2-6 fluorochromes.
When updating panels replacing dimmer fluorochromes such as Pacific Blue with very bright dyes such as Brilliant Violet™ 421, one may want to consider the antigen density of the markers. For example if Pacific Blue is used it is likely on a high antigen density (bright) marker such as CD4. You may want to put a lower antigen density or dimmer marker on the Brilliant Violet BV421 and put CD4 on a dimmer fluorochrome.
Possible moderate spillover into the Alexa Fluor® 700 and PerCP-Cy™5.5
Green (532 nm) and Yellow Green (561 nm) Lasers
The green and yellow green lasers are typically used to detect PE and PE tandems. These dyes are typically brighter off the green/yellow green lasers compared to the traditional blue (488 nm) laser. Use of the green/yellow green laser can facilitate the spread of markers across lasers decreasing compensation requirements.
PE
PE
Relative brightness
+++++
Recommended filter
575/26
Excitation max (nm)
565
Emission max (nm)
575
Recommended for
Low density markers
Incompatible dyes
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers
PE/Dazzle™ 594
PE/Dazzle™ 594
Relative brightness
+++++
Recommended filter
565/26
Excitation max (nm)
565
Emission max (nm)
575
Recommended for
Low density markers
Incompatible dyes
PE-CF594
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers
PE-Cy™5
PE-Cy™5
Relative brightness
+++++
Recommended filter
670/14
Excitation max (nm)
565
Emission max (nm)
670
Recommended for
Low density markers
Incompatible dyes
PerCP, PerCP-Cy™5.5
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers, Tandem dye
PE-Cy™7
PE-Cy™7
Relative brightness
++++
Recommended filter
780/60
Excitation max (nm)
496
Emission max (nm)
785
Recommended for
Low density markers
Incompatible dyes
APC-Cy™7 (on instruments with collinear lasers)
Comments
Compatible with green (532 nm) and yellow-green (561 nm) lasers; Tandem dye is sensitive to degradation. Protect from light and avoid prolonged exposure to paraformaldehyde containing fixatives.
