Why cycle when you can image in a single round?
The power of single-round multiplex IF imaging
Orion™ Spatial Imager gives you 20 channels in a single scan
Multiplexed immunofluorescence (mIF) imaging methods enable researchers to visualize and analyze multiple biomarkers simultaneously within tissue samples. While certain technologies employ multi-cycle staining and imaging workflows—staining and imaging low number of antibodies in sequential rounds—this approach, while effective, comes with inherent challenges that impact data quality and throughput.
RareCyte developed the Orion Spatial Imager to address these limitations head-on. Orion delivers single-round 20-channel multiplex imaging, eliminating the need to cycle antibodies to reach a high plex. Orion is the only single-round mIF imager in the market.
What happens when you repeatedly strip your antibodies?
Cyclic approaches rely on repeated rounds of antibody stripping, staining, and imaging.
This process has a negative effect on:
- Tissue Integrity: Repeated stripping can physically damage the tissue, resulting in sample degradation or shifts between cycles that complicate image alignment and analysis.
- Signal Intensity: Successive rounds of stripping and staining often diminish antibody signal intensity, increasing the risk of data loss. You will need to decide when to apply different antibodies to achieve the best results, adding complexity to panel building.
We work a lot with delicate tissues…so we wanted to make sure that we are never going to lose any part of it, and therefore we wanted to find a way that was going to avoid methods that rely on cycling. Stripping can be quickly harsh on the tissues…and you absolutely need to verify the complete removal [of antibodies]. Because if you don’t, you’re going to have cross reactivity between the antibodies that were left over from previous samples.”
How does cycling impact throughput?
Throughput is a well-recognized challenge for spatial proteomic platforms, especially in large-scale studies and clinical applications where efficiency is critical. Cycling antibodies, even using systems with automated microfluidics, require multiple rounds of staining and imaging, which significantly extends the workflow. In contrast, Orion achieves the same plex and slide coverage in a single scan, making it over 10x faster than other multiplex IF imaging platforms available today.
The catch? Because of its iterative nature, the CycIF workflow is time-consuming, and the tissues can struggle to handle many rounds of this process"
From the new Select Science article:
Advanced spatial biology sheds new light on therapeutic research. Read the article ➝
How Orion achieves high-plex imaging without cycling
Orion's innovative design leverages RareCyte’s proprietary ArgoFluor™ dyes and advanced 20-channel imaging system. Each antibody in the panel is conjugated to a specific ArgoFluor dye, ensuring that each biomarker corresponds to a distinct Orion channel. Orion’s nine-laser illumination system spans a wide wavelength range, allowing Orion to capture 20 channels in a single scan.
ArgoFluor dyes have wide spectral spacing to ensure data quality.
Explore the future of spatial biology
Learn more about the advantages of single round multiplex imaging and how Orion is enabling high-resolution spatial analysis at unmatched speeds.
Register for our latest webinar to learn more about the Orion platform:
Breaking Barriers in Spatial Proteomics with Single-Round Imaging
Mar 18, 2025
9:00 am PST | 12:00 pm EST | 18:00 CET
Tad George, PhD
Sr VP Biology R&D
Join us to explore how the Orion Spatial Imager is transforming spatial proteomics with high-throughput, single-round imaging—eliminating the need for cycling and improving image quality. Unlike traditional multiplex immunofluorescence systems that require iterative staining and imaging, Orion delivers 18 plex in a single scan, streamlining workflows and accelerating discoveries.
In this webinar, you'll get an in-depth look at Orion’s capabilities, a walkthrough of its single-round workflow, and customer case studies showcasing key applications. Don't miss this opportunity to learn how Orion is setting a new standard in spatial proteomics.
