TABLE 1.

Multiplex technology providers and topline methods for spatial biology.

Multiplex tissue imaging providers, and their use of brightfield-, fluorescence- or DNA- and mass cytometry-based methods for spatial biology.

Technology provider	Top-level summary
Agilent a	RUO, Clinical/IVD solutions provider for IHC, sequential brightfield multiplexing
Akoya Biosciences b	Single-cell, sub-cellular RNA and protein detection using tyramide signal amplification with multispectral whole-slide fluorescence imaging
10X Genomics c	Spot-based transcriptomics using release-ligated probe pairs, with in situ protein and multiomics on roadmap
Cell IDx d	Single-cell, sub-cellular protein detection using proprietary anti-hapten antibodies for sequential brightfield and fluorescent multiplexing
IonPath e	Mass spectrometry cytometry with ROI spatial tissue analysis using metal-conjugated antibodies
Leica f	RUO, Clinical/IVD solutions provider for IHC, brightfield and fluorescent multiplexing through partnerships, higher plex fluorescent cyclic multiplexing using dye inactivation methodology
Lunaphore g	High-plex staining platform with proprietary fluidics technology, no reagent solution
Miltenyi h	Flow-centric, moving into spatial proteomics using recombinantly engineered antibody fragments coupled to releasable fluorochromes for sequential fluorescence multiplexing
Nanostring i	Spot-based protein and RNA profiling, using photocleavable oligonucleotide probes, single cell analysis and subcellular resolution on roadmap
Standard BioTools j	Imaging mass cytometry with ROI spatial tissue analysis using CyTOF technology with using metal-conjugated antibodies
Ultivue k	Single-cell, sub-cellular protein detection using DNA barcoded antibodies for sequential fluorescence multiplexing
Ventana l	RUO, Clinical/IVD solutions provider for IHC, predominantly brightfield multiplexing but tyramide/hapten-based signal amplification for fluorescence multiplexing
Markers	2–3	4–5	6–7	8–9	10+

Intended Use m	Translational Diagnostic	Translational Diagnostic	DiscoveryHypothesis Translational	Discovery Hypothesis	DiscoveryHypothesis
Tech Method	Brightfield (absorbance)	Fluorescence (emission)			Barcode/Ion (mass or ID tag)
Example Providers	Agilent, Leica, Ventana	Akoya, Cell IDX, Ultivue, Leica, Ventana			10X Genomics, IonPath, Leica (FL), Miltenyi, Nanostring, Standard BioTools
Phenotypes	Low	Medium	Medium	High	Very High
Dynamic Range	Low—1 Log	Medium—3 Log	Medium—3 Log	Medium—3 Log	High—5 Log
Resolution	200nm, WSI	200nm, WSI	200nm, WSI	200nm, WSI/ROI	1µM, ROI
Throughput	High—0.5 min/mm2	Medium—1 min/mm2	High—1 min/mm2	Low—10 min/mm2	Very Low—1 h/mm2
Build Complexity	Low	Medium	Medium	High	Very High
Providers n	Many	Growing	Growing	Fewer	Few

Assumptions and Notes

^a https://www.agilent.com/en/product/immunohistochemistry

^b https://www.akoyabio.com/

^c https://www.10xgenomics.com/

^d https://cellidx.com/

^e https://www.ionpath.com/

^f https://www.leicabiosystems.com/us/ihc-ish/

^g https://lunaphore.com/

^h https://www.miltenyibiotec.com/US-en/products/macs-imaging-and-microscopy.html

ⁱ https://nanostring.com/

^j https://www.standardbio.com/products-services/technologies/imaging-mass-cytometry

^k https://ultivue.com/

^l https://diagnostics.roche.com/us/en/products/product-category/anatomical-pathology.html

^m For definition, see Table 2: Intended use performance requirements for spatial biology using mIF and MSI.

ⁿ Includes contract research organizations (CRO) offering technology, as additional solution providers.