7-Color Imaging

The 7-Color Challenge

Visualizing four colors usually isn’t challenging with conventional bandpass filters. However, when you exceed four colors, the fluorochromes begin to bleed into each other’s channels.
The complete spectrum, from violet to far-red, can only be divided into a certain number of channels before spillover and non-specific signals arise. To achieve a clean signal, each channel must be narrow enough to prevent cross-talk – in other words, maintain specificity – while still capturing as much emission signal as possible.

We have optimized it for you with our patented SpectraSplit® 7,  a 7-Channel filter set consisting of 4 bandpass filter sets.

Discover the Power of Seven Colors

Any high-throughput scanning method is enhanced by the use of more colors, as each staining cycle conveys more information. SpectraSplit® 7 is a patented set of bandpass filters that features carefully calculated and tested bandwidths. SpectraSplit® 7 effectively separate seven common classes of fluorochromes without the need for spectral unmixing. The system is compatible with AlexaFluor®dyes, as well as traditional FITC/Cy3/Cy5/Cy7 dyes and many varieties of Opal™ dyes. With its user-friendly design, simply choose one fluorochrome from each class to produce 7-color images free from channel spillover. The data generated requires no spectral unmixing or other types of post-processing.You can proceed directly to analysis with your preferred software.

Fluorochromes compatible with SpectraSplit® 7

Channel Traditional dyes StreptaClick® dyes ThermoFisher Biotium ThermoFisher Atto-tec Akoya Ultivue Sirigen/BD Protein
S-Split Blue (375) DAPI AZDye™ 405 Alexa Fluor 405 CF405 Dylight 405 DAPI DAPI BV421 Hoecht
S-Split Cyan (435) CF®430 CF430 Atto425 Opal p.480 BV480 cCFP
S-Split Green (490) FITC AZDye™ 488 Alexa Fluor 488 CF488 DyLight 488 Atto488 Opal 520 Ultivue, FITC eGFP
S-Split Orange (545) Cy3/TRITC AZDye™ 555,
Cyanine 3, Atto™ 542
Alexa Fluor 546 CF555 DyLight 549 Atto542 Opal 570 Ultivue, Cy3 mOrange/
S-Split Red (590) Texas Red AZDye™ 594 Alexa Fluor 594 CF594 DyLight 594 Atto590 Opal 620 mCherry/mRaspberry
S-Split Far-red (650) Cy5/Cy5.5 AZDye™ 647,
Cyanine 5
Alexa Fluor 647 CF647/680 DyLight 649 Atto647/665 Opal 690 Ultivue, Cy5 miRFP703
S-Split Infra-red (740) Cy7 Alexa Fluor 750 CF750 DyLight 750 Atto740 Opal p.780 Ultivue, Cy7

*Cy3, Cy5, Cy7 are registered trademarks of GE Healthcare, Alexa Fluor and DyLight dyes are trademarks of Thermo Fisher Scientific, CF dyes are trademarks of Biotium, Atto dyes are trademarks of Atto-tec, Opal dyes are trademarks of Akoya Biosciences, Brillialt Violet dyes are trademarks of Sirigen/BD

Quality Filters

The filter sets are custom made by Chroma Technologies and manufactured to the high product standards of Chroma Technologies. SpectraSplit® 7 is a patented technology, exclusive to our brand. These filter sets are not available under any other name.

SpectraSplit® 7 maximizes the capacity of any microscope

SpectraSplit® 7 immediately empowers your microscope with seven independent color channels that each generates crystal clear images. SpectraSplit® 7 is designed to work with both standard and scanning fluorescence microscopes and it is compatible with various lighting sources.

SpectraSplit® in the VS200 Research Slide Scanner from Evident

The SpectraSplit® 7 is available as a seven-channel option on the VS200 Research Slide Scanner from Evident. In conjunction with a high-performance microscope, SpectraSplit® 7 enhances your visual capabilities. For more information, please reach out to your Evident sales representative.

We’ve imaged over 2,500 highly-multiplexed slides with the SpectraSplit 7 filters in the past year and the data in every single sample set is crisp and clean. Our users do a large amount of RNAScope staining utilizing the Opal dye set and it’s been a game changer for us to have the SpectraSplit filters that are precisely matched to these dyes. The imaged samples have been a wide range of tissue sections and cultured cells and none of my other microscopes, both filter-based and with spectral detection, match the Opals as well as SpectraSplit 7.

Christina Baer,
Director, SCOPE Imaging Facility,
UMass Chan Medical School,
Worcester MA

Our company Offspring Biosciences supports project teams in the pharma industry with histology-based contract services. Kromnigons SpectraSplit filters have allowed us to significantly expand our capability to perform multiplexed immunofluorescence analyses with excellent separation between the fluorophores. We can highly recommend them.

Anders Dahlstrand,
Managing Director at Offspring

SpectraSplit® 7

SpectraSplit® 7 are patented filter sets that empower fluorescence microscopes with seven color channels.  The channels are distributed over four filter sets and specifically separate the most commonly used fluorochromes in immunofluorescence (see table below). Hence, you can immunostain tissues with up to seven fluorochromes and still visualize each fluorochrome in the sample with SpectraSplit® 7 without bleedthrough. No additional software and no computational spectral unmixing are required. SpectraSplit® 7 can easily be installed in both standard and scanning fluorescence microscopes, and can be run with different popular light sources. See the description below for more information. Request a Quote


StreptaClick® Color labels antibodies with fluorochromes, mimicking directly conjugated antibodies. StreptaClick®-color provides a rapid, robust, and flexible method for attaching fluorochromes to biotinylated antibodies. Once labeled with StreptaClick®, the antibody is stable and can be used in the same manner as antibodies that are directly conjugated with fluorochromes. The click reaction is highly efficient and can be carried out at any volume, concentration, or buffer. StreptaClick®-color is available in four colors: AZDye™ 488, AZDye™ 594, AZDye™ 647, and Atto™ 542. The AZDyes™ are structurally identical to the Alexa Fluor® dyes, and Atto 542 is a particularly bright dye with a spectral similarity to Cy®3. Each color contains 500 μl of StreptaClick® antibody labeling reagent, capable of labeling 50 μg of biotinylated antibody, sufficient for approximately 50 slides. Select your StreptaClick®-Colors:

StreptaClick®-HRP Mutliplex IHC Kit

390 850 
StreptaClick®–HRP antibody labeling reagent and a novel HRP block buffer that efficiently quenches HRP activity between staining cycles. Unlike other buffers, this HRP block buffer does not require boiling between cycles, thus not only reducing the time for each staining cycle but also making it compatible with both FFPE sections and frozen sections, all while preserving tissue morphology. StreptaClick®–HRP is compatible with biotinylated antibodies from any host and on tissues from mice, humans, rats, and other species. Note: While the kit does NOT include TSA Dyes or Signal Amplification buffer, these items are also available for purchase in our online store.