Table 1.
Overview the mfIHC technologies.
| Method Name | Vendor Name |
Sample | Maximal Labeling |
Direct/ Indirect Detection |
Antibody Conjugation |
Stain Removal Method |
Time Consuming (Per Cycle) * |
Resolution |
|---|---|---|---|---|---|---|---|---|
| Stain removal technologies | ||||||||
| MELC (Toponome imaging systems) |
ToposNomos GmbH | FFPE | 100 | Direct | Fluorescent based | Bleaching | two proteins per hour (one tag/one dye per cycle) |
<40 nm |
| SIMPLE | NA | FFPE | 12 | Indirect | Fluorescent based | Alcohol-soluble red peroxidase substrate AEC method |
3 h | 15–20 μm |
| IBEX | NA | Frozen/FFPE | >65 | Direct/indirect | Fluorescent based | LiBH4-based bleaching | Manual ~3.5 h/automated ~1.5 h | 160 nm |
| Fluorophore inactivation technologies | ||||||||
| MxIF | Cell IDx | FFPE | 60 | Indirect | Fluorescent based | Alkaline oxidation chemistry inactivation | 1 h 15 min | 1 μm |
| CycIF | NA | FFPE | 60 | Direct | Fluorescent based | Hydrogen peroxide and light inactivation | ~24 h | 5 μm |
| Chip Cytometry |
Zellsafe™ | Cell suspensions /frozen/FFPE |
60 | Direct | Fluorescent based | Chemical bleaching or light photobleaching | ~1 h | 5 μm |
| Multiplexed signal amplification | ||||||||
| Multiplex modified hapten-based | UltraPlex™ | FFPE | 4 | Indirect | Fluorescent based | Antibody stripping | 2 h | NA |
| TSA | Roche and Akoya Biosciences |
Cell suspensions /FFPE |
9 | Indirect | Fluorescent based | Antibody stripping | 1 h | 0.25–0.9 μm |
| QDs | NA | FFPE | 5 | Direct/Indirect | Fluorescent based | Chemical bleaching | 6 h | Super resolution |
| DNA barcoding technologies | ||||||||
| DEI | NA | FFPE | 8 | Indirect | DNA-barcoding based | NA | 2–3 h | 20 nm |
| CODEX | Akoya Biosciences |
Cell suspensions/frozen/FFPE | 60 | Indirect | DNA-barcoding based | NA | <1 day (Whole slide imaging) |
260 nm |
| Immuno-SABER | NA | Cell suspensions/frozen/FFPE | 10 | Indirect | DNA-barcoding based | NA | 1 h | Super resolution |
| DSP | NanoString | Frozen/FFPE | 96 | Indirect | DNA-barcoding based | NA | 1–2 h | 10 μm |
| InSituPlex® | Ultivue | FFPE | 15 | Indirect | DNA-barcoding based | NA | 5.5 h | NA |
| Mass cytometry | ||||||||
| IMC | Hyperion | Cell suspensions /frozen/FFPE |
>40 | Direct | Metal-based | NA | 2 weeks (0.5 mm × 0.5 mm ROI takes ~3.5 h with a slide scanner) |
1 μm |
| MIBI | Ionpath | Cell suspensions /FFPE |
40–100 | Direct | Metal-based | NA | 2 weeks (Whole slide imaging) |
260 nm |
* Due to different section size, image acquisition time is various, this is a rough estimate.