Table 2.
Comparison of imaging techniques.
| IHC | IF | Multiplex IHC | Imaging mass cytometry | Spatial protein and transcriptomics | |
|---|---|---|---|---|---|
| Example technologies | –/– | –/– | Opal™ | Hyperion™/MIBI™ | Visium/NanoString |
| Number of markers | 1–2 | 3–4 | Up to eight protein markers | >40 protein markers | Protein and gene expression |
| Sample type | FFPE or Fresh frozen | FFPE or Fresh frozen | FFPE or Fresh frozen | FFPE or Fresh frozen | Fresh frozen (Visium) FFPE or Fresh frozen (NanoString) |
| Antibody availability/format | Purified antibody | Purified or fluorochrome-conjugated antibody | Any purified antibody suitable for IHC | Metal-tagged antibodies (commercially available or self-conjugation) | Slide layered with DNA probes |
| Staining | Time: <1 h Cost: low Difficulty: low |
Time: <1 h Cost: low Difficulty: low |
Time: up to 5 days Cost: low Difficulty: low |
Time: up to 2 days Cost: mid-high (dependent on region size for ablation) Difficulty: mid-high (requires specific instrument) |
Time: up to 2 days Cost: high Difficulty: high (various instruments required) |
| Imaging/data collection | Conventional microscope | Immunofluorescent microscope | Mantra™ imaging: manual Vectra® Polaris™ imaging: automated slide scanning |
Hyperion™ ablation: 1 mm2 tissue approx. 2.5 h MIBI™: 1 mm2 tissue approx. 5 h |
Microscope (protein) RNA sequencing |
| Resolution | Dependent on microscope | Dependent on microscope | 200 nm (dependent on microscope) |
1000 nm | 100 µm |