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.

Conducting preliminary experiments before formal batch testing with enzyme-linked immunosorbent assay (ELISA) is a key method to avoid problems such as sample concentrations exceeding the quantification range, matrix interference, excessive well-to-well variation, and waste of precious samples.

The core insight of this review is to reinterpret the tumour‑suppressive functions of p53. Beyond canonical apoptosis‑driven tumour suppression, p53 can also exert biological effects through multiple non‑apoptotic cell‑death pathways including ferroptosis, necroptosis and pyroptosis, which influence tumorigenesis, tissue injury, neurodegenerative disorders and infections.

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.

This article will use Curve Expert software to complete, step by step, data import, standard curve plotting, fit evaluation, and the calculation of concentrations of the samples to be tested.

After obtaining raw OD readings from ELISA assays, standard‑curve fitting and sample concentration conversion are critical to the reliability of experimental data.

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.