Applications

MIST technology works for most cells and tissue sources for information-rich, high-precision investigations.

Human Diseases

Cancer immunotherapy, immunological disorders, neurodegenerative diseases, developmental biology, and many.

Fundamental Biology

Suspension and adherent cells or

solid tissue in study of developmental biology, Immune system, mechanobiology, etc.

Drug discovery

The performance of drugs on cultured cells or tissue can be assessed with single-cell precision by MIST.

Knowledge Center

MIST Linker - superior performance

High-Efficiency Labeling/Conjugation for Staining and Streamlined Workflows

The MIST Linker can be labeled with fluorophores or conjugated with oligo DNAs. Owing to its high capacity for carrying biotin-tagged oligos and fluorophores, it delivers immunofluorescence (IF) staining intensity comparable to conventional, time-consuming two-step methods. Once conjugated, the antibodies can be used directly for tissue staining.

Purer Antibody-DNA Conjugates

Typically, tissue staining with fluorophore-labeled antibody-DNA conjugates suffers from a low signal-to-noise ratio. This occurs because residual unreacted DNA tends to bind non-specifically to cell nuclei, even when using salmon sperm DNA as a blocking agent. The MIST Linker Cleavable Antibody-DNA Conjugation Kit solves this issue. By combining MIST Quencher™ with a 100 kDa purification filter, the kit yields a relatively purified antibody conjugate that significantly reduces non-specific background staining.

Robust Performance in Multiplexed Cyclic IF

These antibody-DNA conjugates enable 9 cycles of cyclic immunofluorescence (cyclic IF) in our test where we don't need be concerned about whether the antibodies are from the same species or not. Each cleavage step removes more than 90% of the fluorescence signal. During development, we tested adding three tandem photocleavable linkers between the antibody and the fluorophore. This modification did not significantly improve the removal of residual fluorescence, indicating that diffusion into the bulk solution after cleavage rather than cleavage efficiency itself is the limiting factor.

To ensure complete signal removal, applying a mild quenching solution (4% H2O2 and 25 mM NaOH in PBS) for 30 minutes entirely eliminates the remaining fluorescence. This step is particularly helpful for accurately detecting low-abundance proteins in subsequent cycles

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2. Spatial MIST copy-pastes proteins from tissue.

cytoMIST technology is likely the only platform in the spatial proteomics field that copies protein distribution and abundance directly onto a microarray. Consequently, whether analyzing a single target or dozens of plexes, the workflow requires the same amount of time—only about 15 minutes in addition to standard tissue processing and staining. This efficiency becomes especially apparent during high-multiplex assays.

cytoMIST also enables rapid detection of more than 100 proteins at sub-10 μm cellular resolution in any tissue section. The core components of this technology are a highly compact barcode DNA array and custom-designed, UV-cleavable single-stranded (ss) DNA-antibody conjugates. The array contains approximately 50 million barcode-bearing microbeads within a 1 cm x 1 cm area on a glass slide, or 400 million beads in a 2 cm x 4 cm area. The conjugates convert protein detection into DNA detection, allowing researchers to leverage DNA-based techniques for multiplexed analysis. Even without retaining spatial information, cytoMIST has achieved high-throughput analysis of up to 465 proteins per cell—the highest multiplexity achieved to date in single-cell analysis. It has also been used to map the spatial distribution of biomarkers in tissue sections, co-detecting up to 125 proteins.

To capture spatial information during single-cell functional proteomic analysis, cytoMIST transfers, or "prints," protein information from a tissue slice directly onto the array. For example, a mouse brain sample is first stained with antibody-barcode DNA conjugates; the DNA tags are then released and transferred to the MIST array. This transfer process takes just 5 minutes, with signal diffusion limited to a mere 1.2 μm. System sensitivity reaches approximately 180 proteins per cell. Furthermore, cytoMIST technology allows for multiple consecutive rounds of printing from the same tissue section onto a series of MIST arrays to scale up multiplexing.

The accompanying MIST Explorer software provides all the tools necessary to process cytoMIST data. Learn more on this page.