Pathological Research

​Conventional immunohistochemistry (IHC) only enables detection of 1–2 proteins per tissue section, providing merely cell counts while losing in situ spatial information and protein co‑expression profiles. Standard single‑plex immunofluorescence also fails to resolve complex immune suppressive networks. Multiplex immunofluorescence (mIF) allows simultaneous detection of multiple targets on a single FFPE paraffin section, preserves native tissue architecture, and supports cell phenotyping, cell density quantification and spatial distance analysis, making it a key technique for translational tumor immunology research.

From the purple bacterial bodies in Gram staining, to the clearly visible stained cells in cell experiments, and to the easily observable cell colonies in cloning experiments, crystal violet has become one of the common staining reagents in the laboratory due to its good coloring ability. But did you know? Despite all being called 'crystal violet staining,' the staining purpose, operation methods, and result interpretation can be completely different in different experiments. Today, let's get to know crystal violet staining solution, see what it can actually do, and explore the details worth paying attention to in experiments.

Picrosirius Red Staining is a special staining method commonly used to show collagen fibers in tissues. It not only helps observe the distribution of collagen fibers, but can also be combined with a polarized light microscope to further examine the birefringent characteristics of collagen fibers. So, what exactly are the characteristics of Picrosirius Red Staining? How does it differ from the commonly used Masson's Trichrome Staining?

In cell culture, cell passaging and cell counting experiments, besides cell concentration, one key parameter needs to be monitored — cell viability. How many cells in the suspension retain intact plasma membranes? How many have lost membrane integrity? How can we quickly assess cell quality before seeding, passaging or downstream assays? The Trypan Blue Exclusion Assay is a classic, straightforward method for cell viability assessment.

Alcian Blue Staining is a special histochemical staining method widely used to detect acidic mucins. It stains target components blue and facilitates observation of mucin distribution and associated pathological changes in tissue sections.

The article uses a multiplex immunofluorescence (mIF) dataset of malignant pleural mesothelioma as a demonstration case, systematically reviewing the paradigm of tumor microenvironment spatial modeling analysis for multiplex immunofluorescence images, focusing on the applicable conditions and potential methodological pitfalls of various spatial statistical tools, and providing a practical framework that can be referenced for tumor immunology translational research.

H&E staining uses two dyes with selective coloring ability to differentially mark various chemical components within the tissue, establishing a stable, reproducible, and globally applicable tissue coloration system. In principle, all surgical biopsies and resection specimens are first observed with HE-stained sections; techniques such as immunohistochemistry, special staining, and molecular pathology testing are all based on the morphological assessment of HE sections. Therefore, HE staining is not a simple coloring procedure, but a universal morphological language in the field of pathology.

Using pre-treatment core needle biopsy specimens, this study breaks the conventional paradigm limited to cell abundance quantification. It characterizes immune cell composition and spatial proximity interactions, aiming to uncover the mechanisms underlying divergent therapeutic responses under equivalent TIL levels and identify potential immune biomarkers for patient stratification prior to neoadjuvant chemotherapy for TNBC.

Pathological staining is absolutely not the same as ordinary pigment coloring processes. It is a core technology for visualizing microscopic tissue information that is indispensable in clinical pathological diagnosis, histological research, and biological experiments. Its main function is to convert hidden microscopic structural information in transparent sections, which cannot be recognized by the naked eye or under a microscope, into visual optical signals that are clearly layered, significantly different, accurately identifiable, and objectively analyzable through specific color reactions.

Using multiplex immunohistochemistry, the study dissected TIME features of gastric cancer from both immune‑cell density and spatial distribution dimensions and constructed a multi‑dimensional tumor‑infiltrating immune‑cell predictive model, aiming to address the clinical challenge of accurate efficacy prediction for gastric cancer immunotherapy.