Innovation in Spatial Biology: Next-Generation Protein Imaging and Its Applications

Orion Spatial Biology: The Next-Generation Imaging Platform

Author:RareCyte, Inc.

What is Spatial Biology?

Spatial biology is a field of study that explores how cells, genes, and proteins are arranged and interact within and between tissues. By using advanced technologies, researchers can gain a comprehensive understanding of cellular and tissue functions.

Tissues are composed of different types of cells, each with unique functions and conditions, and their spatial arrangement can affect a patient’s health. However, challenges such as overlapping fluorescence, autofluorescence, and slow processing times due to extended staining and data acquisition have hindered progress in this field.

Innovations with Orion Technology

Orion technology overcomes these challenges by providing fast, easy, and multiplexed whole-slide tissue analysis.

  • Single staining and imaging process: Ultra-fast, cost-effective, and preserves tissue
  • Whole-slide imaging on standard glass slides
  • H&E staining at the same location for morphology and protein alignment
  • Design hundreds of panels with flexibility and ease

With Orion technology’s ultra-fast processing and low cost, research speed and usage frequency increase tenfold, enabling the collection of large-scale statistical data in pharmaceutical and clinical trials.

Case Study: Colorectal Cancer Immunotherapy

The clinical trial on colorectal cancer immunotherapy conducted by Dr. Pashtoon Kasi at Weill Cornell Medicine utilized Orion. This study employed 13-color multiplex staining to analyze cell counts, cell states, and spatial context. The results revealed a significant infiltration of immune cells and reduction in tumor size following immunotherapy.

  • Objective: Uncover the mechanism of immunotherapy drugs on colorectal tumor reduction pre-surgery
  • Samples: Biopsy samples from 12 patients before treatment and surgical samples post-treatment
Immunotherapy sample

  • Test: 13-marker multiplex staining with Orion and subsequent quantitative image analysis to assess cell count, cell state, and spatial context
  • Results: Significant immune cell infiltration and tumor shrinkage were observed
Immunotherapy results

Orion Workflow

The Orion workflow enables high-throughput whole-slide analysis, processing dozens of slides per week, with multiple tissue sections or large tissues placed on a single slide.

  1. Sample Preparation: Place samples on standard glass slides; processing area is up to 10 cm²
  2. Use standard IHC/IF protocols for FFPE and fresh frozen samples. Autofluorescence is cleared with UV and light boxes.

Staining

  1. After a single round of staining with Orion reagents, mount the cover glass and let it sit overnight.
  2. ※ Can be stored at -80°C for up to 1 month.

Scanning

  1. Orion scans 18 channels at a rate of 75 minutes per cm²
  2. Remove the cover glass, perform H&E staining, and rescan

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Flexible Panel Design

RareCyte’s Orion reagent portfolio includes hundreds of panels covering over 30 tissue types and more than 50 diseases. Custom biomarkers are labeled using the ArgoFluor dye kits and validated against IHC. These reagents are small molecules, easily conjugated to antibodies via amine groups, making them simple to use. Panels can be designed in as little as two weeks.

Conclusion

Orion enables the processing of large-scale samples, accommodating large tissues or multiple tissue sections on a single slide, processing dozens of slides weekly, and supporting large-scale cohort studies in clinical research or shared instrumentation environments with numerous users.

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