Pathological Research

In recent years, across research fields such as tumor immunology, infection and inflammation, and neuroscience, an increasing number of reviewer comments have begun to feature a common keyword — "insufficient spatial information". Many researchers are puzzled: they have completed Western blot, qPCR, and even single-cell sequencing, yet are still asked to supplement spatial-level evidence. The core issue lies in an important transformation that modern life sciences are undergoing: the research focus is no longer just "whether something is expressed" or "how much is expressed", but "where it is expressed" and "who it coexists with". This cognitive leap from molecules to structures has gradually made spatial information a key dimension in the paper evaluation system, and TSA multiplex immunofluorescence technology has emerged as an important tool connecting molecular and tissue structures in this context.

Unlike traditional fluorescent immunostaining, the TSA system does not simply enhance fluorescence intensity, but achieves "tissue deposition" of signals through enzymatic catalysis, fundamentally changing the signal existence form and detection logic, thereby supporting tissue in-situ detection systems with up to 5-8 colors or even higher dimensions.

Today, we will provide you with a comprehensive understanding of this revolutionary technology from the perspectives of technical principles, experimental workflow, application cases, and advantages analysis.

In modern life science research, the complexity of tissue microenvironments poses higher demands on scientific research. Multiplex immunofluorescence (mIHC) technology, with its advantages of simultaneously detecting multiple targets and preserving spatial information, has become an indispensable tool in fields such as tumor immunology, neuroscience, and stem cell research.

Over the past few decades, immunohistochemistry (IHC) has been the standard tool for tumor tissue research: from HER2, Ki-67 to PD-L1, it has allowed researchers and pathologists to clearly see the distribution of individual proteins in tissues. However, as tumor research enters the "spatial biology era," information from single markers is far from sufficient.

This study combined sequential stepwise staining with tyramide signal amplification detection technology, successfully establishing a multiplex immunofluorescence staining system suitable for fully automated staining platforms, breaking through the technical barrier that traditional automated staining cannot use same-species primary antibodies for multi-target detection.

What are the differences between traditional IHC and mIHC? Why are more and more studies choosing mIHC?

In recent years, Multiplex Immunohistochemistry (mIHC), with its ability to simultaneously detect multiple markers and analyze spatial information, has become an important tool for tumor microenvironment research, immunotherapy evaluation, and spatial pathology studies. However, compared to traditional IHC, the mIHC experimental workflow is more complex. From antibody screening to Panel design, to TSA staining and image analysis, every step can affect the final result.

With the rapid development of tumor microenvironment research, spatial biology, and precision pathological diagnosis, Multiplex Immunofluorescence (mIHC) has become an important tool for tissue sample analysis. Compared with traditional immunohistochemistry (IHC) and conventional immunofluorescence (IF), mIHC enables simultaneous detection of multiple protein markers on the same tissue section, allowing cell phenotype identification, spatial localization analysis, and cell-cell interaction studies.

mIHC, short for Multiplex Immunohistochemistry, is a high-throughput, high spatial resolution pathological detection technology iteratively upgraded from traditional immunohistochemistry (IHC).