Table 1.

Differences and comparisons of the various characteristics among the different multiplex platforms and assays.

Assay	D.Ultra [32]	MICSSS [33]	Vectra/Polaris [35]	Opal [39]	CODEX [41,42]	InSituPlex [43]	GeoMx DSP [44,45]	MIBI-TOF [46,47]	Cy-TOF [48]
Vendor	Roche	Remark et al.	Akoya Biosciences	Roche and Akoya Biosciences	Akoya Biosciences	Ultivue	Nanostring	IonPath	Bodenmiller et al.
Antibody Conjugate	chromogen	chromogen	fluorescent	Opal fluorescent	DNA barcode	DNA barcode	DNA barcode	Metal isotope	Metal isotope
Tissue disruption	no	no	no	no	no	no	no	Yes (adjustable)	yes
Biomarkers	5	10+	∼9	∼8	40+	∼5	70+	30+	30+
Staining cycles	Iterative	Iterative	Iterative	Iterative	Iterative	1	1	1	1
Scanning	camera	camera	Vectra Polaris	Vectra Polaris	CODEX fluidics	compatible with various systems	GeoMx	TOF spectrometer	TOF spectrometer
Profiling area	Whole slide	Whole slide	adjustable (660 $\mathsf{\mu }$m${}^2$ → whole slide)	adjustable (660 μm${}^2$ → whole slide)	660 $\mathsf{\mu }$m${}^2$	NA	adjustable (280 μm${}^2$ → 800 $\mathsf{\mu }$m${}^2$)	800 $\mathsf{\mu }$m${}^2$	1 mm${}^2$
Software	NA	NA	InForm	InForm QuPath	CODEX analysis manager & viewer	compatible with various systems	GeoMx data analysis suite	MIBIAnalysis	HistoCat, Cell Profiler
Resolution	Human vision	Human vision	0.25–0.9 $\mathsf{\mu }$m	0.25–0.9 $\mathsf{\mu }$m	0.26 $\mathsf{\mu }$m	NA	10 $\mathsf{\mu }$m	0.26 $\mathsf{\mu }$m	1 $\mathsf{\mu }$m
Spatial information	yes	yes	yes	yes	yes	yes	limited	yes	yes
Single-cell information	yes	yes	yes	yes	yes	yes	limited	yes	yes
Disadvantages	Human Vision Limits, Semi- Quantitative	time- consuming, coverslip removal errors	costly, limited marker numbers	costly, limited marker numbers	costly, time- consuming, difficult cell segmentation, difficult panel validation	limited marker numbers	costly, limited spatial information, limited single cell information	special training, costly, time- consuming, difficult cell segmentation, difficult panel validation	special training, costly, time- consuming, difficult cell segmentation, difficult panel validation