Laboratory Investigation最新文献

{"title":"Deep Learning–Guided Quantitative Analysis Establishes Optimized BRAF V600E Immunohistochemical Criteria for Colorectal Cancer: A Multiplatform Validation Study","authors":"Yehan Zhou ,&nbsp;Jiayu Li ,&nbsp;Chengmin Zhou ,&nbsp;Jun Hou ,&nbsp;Jieyu Wang ,&nbsp;Ting Lan ,&nbsp;Dan Wan ,&nbsp;Yuan Tu ,&nbsp;Yungchang Chen ,&nbsp;Qiao Yang ,&nbsp;Jincheng Luo ,&nbsp;Dan Luo ,&nbsp;Lin Shi ,&nbsp;Yang Liu","doi":"10.1016/j.labinv.2025.104215","DOIUrl":"10.1016/j.labinv.2025.104215","url":null,"abstract":"<div><div>Accurate detection of BRAF V600E mutation is critical for guiding therapeutic strategies. Unlike other solid tumors, colorectal cancer (CRC) lacks reliable immunohistochemical (IHC) interpretation criteria. This study aimed to establish CRC-specific IHC criteria through quantitative analysis. A cohort of 250 CRC cases with paired IHC and genetic testing (qPCR and next-generation sequencing) results was analyzed. Cross-platform generalization capability of 3 BRAF V600E antibodies was validated. Previously reported IHC criteria were applied and discordant cases were analyzed. A deep learning–based digital pathology platform quantified IHC parameters (H-score, staining intensity, and percentage). Receiver-operating characteristic analysis identified optimal thresholds, which were translated into practical criteria. External validation was performed to confirm generalizability. Cross-platform validation revealed consistent antibody performance across platforms, with absorbance optical density (2.0-2.3) and H-scores (145-160) showing no significant intergroup differences (<em>P</em> &gt; .05). Initial comparison of existing criteria demonstrated 80.4% to 84.8% concordance with molecular testing. Discordant cases exhibited 5 distinct abnormal staining patterns. Artificial intelligence–driven quantification identified H-score 52.675 as the optimal upper cutoff (area under the curve [AUC], 0.938), translated into a positive criterion of &gt;25% 2+ or &gt;15% 3+ stained cells. A negative criterion of &lt;20% 1+ cells was established. Cases with atypical staining patterns required molecular confirmation. The optimized criteria achieved superior concordance in internal (AUC, 0.932) and external validation (AUC, 0.977). This study established refined BRAF V600E IHC criteria for colorectal cancer using precision quantitative analysis. The optimized protocol significantly improves accuracy and standardization in complex real-world scenarios, demonstrating strong potential for broad clinical adoption.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104215"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Expression of Homo Sapiens (Hsa)-miR-139-5p as a Clinically Feasible Prognostic Marker for Differentiated Thyroid Cancer","authors":"Natalia Martínez-Puente ,&nbsp;Ignacio Ruz-Caracuel ,&nbsp;Luis J. Leandro-García ,&nbsp;Héctor Pian-Arias ,&nbsp;Zaira Vega-Corral ,&nbsp;Rocío Letón ,&nbsp;Roberta Radu ,&nbsp;Mariolga Berrizbeitia ,&nbsp;Sara Mellid ,&nbsp;Clara Reglero ,&nbsp;Milton E. Salazar-Hidalgo ,&nbsp;Ester Arroba ,&nbsp;Alberto Díaz-Talavera ,&nbsp;Mónica Marazuela ,&nbsp;Amparo Benito-Berlinches ,&nbsp;Irene González-García ,&nbsp;Sandra Campos-Mena ,&nbsp;María D. Lozano-Escario ,&nbsp;Sonsoles Guadalix ,&nbsp;María Calatayud ,&nbsp;Cristina Montero-Conde","doi":"10.1016/j.labinv.2025.104199","DOIUrl":"10.1016/j.labinv.2025.104199","url":null,"abstract":"<div><div>Prognostic uncertainty often leads to overtreatment of differentiated thyroid cancer (DTC) or unspecific management of more aggressive entities. MiR-139-5p (miR-139-5p) has emerged as a promising prognostic factor that may enhance current individual risk assessment systems. Therefore, we aimed to validate miR139-5p expression as a prognostic marker in DTC using a standardized method and to establish expression cutoff values for discriminating prognostic groups. In addition, we explored an in situ approach to analyze this microRNA expression as a potential molecular tool for clinical practice. We collected a tissue series of 132 samples, including thyroid tumors, adjacent normal thyroid tissue, and lymph node metastases from a long-term follow-up retrospective cohort of 60 patients with DTC with either progressive/persistent disease or an excellent response to primary treatment. We first identified recurrent tumor driver mutations and <em>TERT</em> promoter mutations using next-generation sequencing. Through a standardized paired tumor/normal qPCR analysis, we confirmed a significant reduction in miR139-5p expression in progressive/persistent DTCs compared with excellent response DTCs (<em>P</em> value = .002). Further analysis, including thyroid cancer The Cancer Genome Atlas tumor/normal pairs (n = 59), showed a strong association between reduced miR139-5p expression and <em>TERT</em> promoter mutations (<em>P</em> &lt; .001), as well as advanced local or distant metastasis at diagnosis (<em>P</em> = .031). Next, we established miR139-5p^HIGH and miR139-5p^LOW tumor/normal cutoff values to discriminate prognostic groups, with high expression predicting excellent response and low expression predicting disease progression/persistence. Cutoff values were defined through logistic regression and receiver operating characteristic curve analysis and validated in an independent cohort (n = 38). Quantitative image analysis using QuPath software of an automatic chromogenic in situ hybridization assay for miR139 detection further supported the qPCR findings and revealed heterogeneous intratumor miR139 expression, which was lowest in the Ki-67 proliferation index--positive foci. Overall, our data indicate that miR139 expression assessment is a feasible tool for clinical use, potentially reducing overtreatment during primary DTC interventions and supporting a risk-adjusted follow-up schedule.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104199"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144181725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinicopathologic, Cellular, and Molecular Analyses of Pulmonary Neuroendocrine Carcinoma With High Expression of Hepatocyte Nuclear Factor 4 Alpha","authors":"Kei Asayama ,&nbsp;Ryota Matsuoka ,&nbsp;Suzuka Tachi ,&nbsp;Aya Shiba-Ishii ,&nbsp;Yoshihiko Murata ,&nbsp;Tomoki Nakagawa ,&nbsp;Yosuke Furuhashi ,&nbsp;Hitomi Kawai ,&nbsp;Ayako Suzuki ,&nbsp;Yutaka Suzuki ,&nbsp;Naohiro Kobayashi ,&nbsp;Yukio Sato ,&nbsp;Nobuyuki Hizawa ,&nbsp;Yoshinori Murakami ,&nbsp;Toshiro Niki ,&nbsp;Daisuke Matsubara","doi":"10.1016/j.labinv.2025.104210","DOIUrl":"10.1016/j.labinv.2025.104210","url":null,"abstract":"<div><div>Pulmonary neuroendocrine carcinoma (NEC), including small cell carcinoma (SCLC) and large cell neuroendocrine carcinoma (LCNEC), is highly aggressive and has a poor prognosis. The molecular subtyping of NECs has recently attracted attention, and we identified a new NEC subtype, the hepatocyte nuclear factor 4α (HNF4α) subtype. HNF4α, a transcription factor associated with gastrointestinal differentiation, and TTF-1 are mutually and exclusively expressed in lung adenocarcinomas; however, the characteristics of HNF4α-high NEC and TTF-1-high NEC have yet to be compared. We immunohistochemically examined the characteristics of HNF4α-high NEC in 83 surgically resected specimens (37 SCLCs and 46 LCNECs) and revealed that HNF4α-high and TTF-1-high NEC accounted for 15% (12/83) and 47% (39/83), respectively. In SCLCs, HNF4α-high cases (n = 3) and TTF-1-high cases (n = 20) were almost confined to the neuroendocrine phenotype with high ASCL1 expression, and the expressions of HNF4α, TTF-1, and POU2F3 were mutually exclusive. Similar results were obtained for LCNECs; however, some HNF4α-high cases were positive for TTF-1 or YAP1, possibly due to the heterogeneity of LCNEC. Therefore, we investigated the heterogeneity of LCNEC and performed a spatial transcriptome analysis of 1 HNF4α-high LCNEC case, which revealed a mutually exclusive mixture of different subgroups characterized by HNF4A and NKX2-1 (TTF-1) expressions. A whole-genome analysis of 10 LCNECs showed that <em>NFE2L2/KEAP1</em> mutations were characteristic of HNF4α-positive LCNECs. A prognostic analysis revealed a significantly worse prognosis in HNF4α-high LCNECs than in HNF4α-low LCNECs. A cell line analysis showed that TTF-1-high-expressing (Lu139/H889/H510A) and HNF4α-high-expressing (VMRC-LCD/H810) lines were consistent with ASCL1-high-expressing lines. HNF4α knockdown/knock-in experiments in VMRC-LCD and SBC5 (HNF4α-negative) revealed that HNF4α promoted cell proliferation by inhibiting apoptosis. The HNF4α-subtype of pulmonary NEC is a unique subtype, characterized by a neuroendocrine phenotype with high ASCL1 expression and mutual exclusivity with the TTF-1/POU2F3 subtypes. <em>NFE2L2/KEAP1</em> mutations and HNF4α itself are potential therapeutic targets for this subtype.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104210"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144564870","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Validation Workflow for Novel Oligonucleotide Sequences to Expand the Multiplexing Capacity of the CO-Detection by indEXing (CODEX) Platform","authors":"Alexander Rochwarger ,&nbsp;Louisa Kaufmann ,&nbsp;Jing Zhao ,&nbsp;Ahmad Makky ,&nbsp;Nhat Anh Nguyen ,&nbsp;Nastassja Lehmann ,&nbsp;Nikolay Samusik ,&nbsp;Christine Beschorner ,&nbsp;Saumya S. Manmadhan ,&nbsp;Karen Greif ,&nbsp;Christian M. Schürch","doi":"10.1016/j.labinv.2025.104200","DOIUrl":"10.1016/j.labinv.2025.104200","url":null,"abstract":"<div><div>Antibody-based multiplexed tissue imaging has the potential to provide critical advances in biological discoveries and their translation for clinical applications. With the increasing introduction of markers and modalities for spatial analysis, there is an according demand for the expansion of multiplexing capacities of such imaging platforms. CO-Detection by indEXing (CODEX) is a widely used multiplexed tissue imaging platform that utilizes DNA-conjugated antibodies for imaging. The multiplexing capacity of CODEX is limited by the availability of unique DNA-oligonucleotide sequences for antibody barcoding. In this study, we demonstrate a workflow for the validation and the introduction of novel sets of candidate DNA-oligonucleotide sequences for CODEX. Through cross-validation multicycle experiments with the already published library of DNA barcodes, we here present a set of 27 novel oligonucleotide sequences for CODEX, increasing the potential multiplexing capacity to 85+ markers. We confirmed the utility of the new barcodes using a 74-plex antibody panel on a multitumor tissue microarray of paraffin-embedded tissues. The workflow presented here provides a reproducible method for extending the plexity of the CODEX platform, facilitating a deeper understanding of tissue microenvironments.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104200"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiscale Fusion Models With Genomic, Topological, and Pathomic Features to Predict Response to Radiation Therapy for Non–Small Cell Lung Cancer Patients","authors":"Yu Jin ,&nbsp;Hidetaka Arimura ,&nbsp;Takeshi Iwasaki ,&nbsp;Takumi Kodama ,&nbsp;Noriaki Yamamoto ,&nbsp;Yunhao Cui ,&nbsp;Yoshinao Oda","doi":"10.1016/j.labinv.2025.104204","DOIUrl":"10.1016/j.labinv.2025.104204","url":null,"abstract":"<div><div>Artificial intelligence models with biomarkers to predict treatment responses to radiation would be necessary to maximize the treatment outcomes of individual patients, especially with histopathology images routinely obtained before treatment. We hypothesized that multiscale features, such as genomic (GM), pathomic (PM), and topological (TP) features, could be associated with the radiation response. We investigated fusion models with multiscale features in histopathology images to predict response to radiation therapy for patients (responders) with non–small cell lung cancer. Ten radiosensitivity-related (radiosensitive and radioresistant) genes were deployed as GM features. PM features were extracted from histopathology images by conventional PM analyses. TP features represent the intrinsic properties of tumor cells using Betti numbers, which are mathematical invariants. We analyzed non–small cell lung cancer patients from The Cancer Genome Atlas and Clinical Proteomic Tumor Analysis Consortium who received radiotherapy and established 3 base models with GM, TP, and PM features, respectively, and 3 fusion models. The TP model showed a higher area under the receiver operating characteristic curve of 0.707 (<em>P</em> = .026, log-rank test in overall survival analysis) in the internal test data set and 0.720 (<em>P</em> = .136) in the external test data set. The results indicated that the TP models achieved better classification and prognostic prediction powers than the other base models. The inner-cell TP structure may have the ability to reveal the cell radiosensitivity-related information. Furthermore, the best fusion model with GM, TP, and PM features achieved the highest area under the receiver operating characteristic curve of 0.846 (<em>P</em> = .019) and 0.731 (<em>P</em> = .043) in predicting the treatment response and prognoses in the internal and external test data sets, respectively. This study demonstrated the predictive power of the multiscale fusion model for histopathology images, which may assist clinical physicians in the selection of responders to radiation for personalized radiation therapy and would be substantially beneficial for patients with cancer.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104204"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144293991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Histopathological Insights into Metastatic Breast Cancer Gained From Rapid Autopsies","authors":"Gitte Zels ,&nbsp;Anirudh Pabba ,&nbsp;Maxim De Schepper ,&nbsp;Tatjana Geukens ,&nbsp;Karen Van Baelen ,&nbsp;Marion Maetens ,&nbsp;Amena Mahdami ,&nbsp;Sophia Leduc ,&nbsp;Edoardo Isnaldi ,&nbsp;Ha-Linh Nguyen ,&nbsp;Imane Bachir ,&nbsp;Josephine Van Cauwenberge ,&nbsp;Kristien Borremans ,&nbsp;Birgit Weynand ,&nbsp;Elia Biganzoli ,&nbsp;Patrick Neven ,&nbsp;Hans Wildiers ,&nbsp;Wouter Van Den Bogaert ,&nbsp;François Richard ,&nbsp;Christine Desmedt ,&nbsp;Giuseppe Floris","doi":"10.1016/j.labinv.2025.104202","DOIUrl":"10.1016/j.labinv.2025.104202","url":null,"abstract":"<div><div>The biology of metastatic breast cancer is poorly understood, and its understanding is hampered by limited access to metastatic tissue. Post-mortem tissue donation programs may represent a step forward to circumvent this problem, allowing access to large volumes of samples that would often be inaccessible otherwise. In this context, we have set up the UZ/KU Leuven Program for post-mortem Tissue Donation to Enhance Research (UPTIDER, NCT04531696). In this study, we performed detailed histopathological examination of 662 unique metastases collected during autopsy from the first 20 patients in our UPTIDER program. Tissue procurement was guided by a patient-specific tissue donation plan based on available clinical and imaging data. Central pathology review included revision of the primary tumor and biomarker assessment (estrogen receptor [ER], progesterone receptor [PR], human epidermal growth factor receptor 2, and KI67). Linear mixed quantile regression was used to assess relevant associations. Metastases in bones, liver, pleura, and nonaxillary lymph nodes were present in up to 17 patients. Our major findings include (1) an important clinical underestimation of disease burden in patients with invasive lobular carcinoma; (2) a relatively modest disease burden associated with leptomeningeal metastases; (3) a higher than anticipated loss of predictive biomarkers in metastases from primary ER- positive and/or PR positive tumors (up to 84% and 100% of the patients having at least 1 ER-negative or PR-negative lesion, respectively); (4) a high variability in KI67% between metastases with frequent zonation pattern; and (5) a high frequency of metastases with human epidermal growth factor receptor 2–low or –ultralow status. Despite the challenging setup of UPTIDER, we demonstrated here that the data and observations that emerge from this program have high potential for clinical translatability.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104202"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144191945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular Subtypes of Balanopreputial and Urethral Male Genital Lichen Sclerosus: Distinct Transcriptomic and Clinicopathological Profiles","authors":"Xianjie Xiu ,&nbsp;Zhenwei Yu ,&nbsp;Georgios Kravvas ,&nbsp;Christopher B. Bunker ,&nbsp;Liang Cheng ,&nbsp;Guangyu Mao ,&nbsp;Juan Tang ,&nbsp;Ruihang Zhang ,&nbsp;Tianzheng Hao ,&nbsp;Lichun Yang ,&nbsp;Zeyu Wang ,&nbsp;Weidong Zhu ,&nbsp;Wei Yuan ,&nbsp;Zuojing Yin ,&nbsp;Lujie Song","doi":"10.1016/j.labinv.2025.104206","DOIUrl":"10.1016/j.labinv.2025.104206","url":null,"abstract":"<div><div>Male genital lichen sclerosus (MGLSc) is a heterogeneous and aggressive disease characterized by varying severities of balanopreputial and urethral disease (MGLSc-US) and outcomes, including stricture. This study aims to elucidate the transcriptomic heterogeneity of MGLSc and explore its associations with histological and clinical features. We collected 40 preputial samples and 14 urethral tissue samples from patients with MGLSc-US, non-MGLSc urethral strictures, and redundant prepuce. Bulk RNA sequencing was performed to comprehensively profile the transcriptome. Molecular subtypes, functional features, and gene signatures were identified in MGLSc prepuce and urethral lesions. Additionally, we examined the histological and clinical features specific to each subtype. Two distinct transcriptomic subtypes in preputial lesions were identified. Subtype 1 was characterized by the upregulation of immune pathways and increased lymphocytic stromal infiltration. Subtype 2 showed an upregulation of epithelial cell proliferation and cellular stress response pathways. Both subtypes demonstrated features of hyperkeratosis; however, atrophy was specifically associated with subtype 1, whereas subtype 2 showed significant downregulation of extracellular matrix organization pathways and milder dermal sclerosis. <em>PLEK</em>, <em>PIK3AP1</em>, <em>NCF1</em>, <em>CTSS</em>, and <em>SELL</em> and <em>EVPL</em>, <em>RAPGEFL1</em>, and <em>TMEM79</em> were identified as 2 subtype gene signatures across preputial and urethral lesion cohorts. Clinically, subtype 2 was significantly associated with longer US segments compared with subtype 1. This study provides the first detailed transcriptomic characterization of MGLSc, identifying 2 distinct molecular subtypes with stratified markers. These findings offer a foundation for clinical and molecular classification of MGLSc and may guide management strategies and novel therapeutic developments for this challenging condition.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104206"},"PeriodicalIF":5.1,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144340253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Antibody-Based Multiplex Image Analysis: Standard Analytical Workflows and Artificial Intelligence Tools for Pathologists","authors":"Mohamed Omar ,&nbsp;Giuseppe Nicolo’ Fanelli ,&nbsp;Fabio Socciarelli ,&nbsp;Varun Ullanat ,&nbsp;Sreekar Reddy Puchala ,&nbsp;James Wen ,&nbsp;Alex Chowdhury ,&nbsp;Itzel Valencia ,&nbsp;Cristian Scatena ,&nbsp;Luigi Marchionni ,&nbsp;Renato Umeton ,&nbsp;Massimo Loda","doi":"10.1016/j.labinv.2025.104220","DOIUrl":"10.1016/j.labinv.2025.104220","url":null,"abstract":"<div><div>Conventional histopathology has traditionally been the cornerstone of disease diagnosis, relying on qualitative or semiquantitative visual inspection of tissue sections to detect pathological changes. Singleplex immunohistochemistry (IHC), although effective in detecting specific biomarkers, is often limited by its single-marker focus, which constrains its ability to capture the complexity of the tissue environment. The introduction of multiplexed imaging technologies, such as multiplex IHC and multiplex immunofluorescence, has been transformative, enabling the simultaneous visualization of multiple biomarkers within a single tissue section. These approaches complement morphology with quantitative multimarker data and spatial context, providing a more comprehensive view of cellular interactions and disease mechanisms. However, the rich data from multiplex IHC/multiplex immunofluorescence experiments come with significant analytical challenges, as large multichannel images require comprehensive processing to transform raw imaging data into quantitative and meaningful information. This review focuses on the standard digital image analysis workflow for multiplex imaging in pathology, covering each step from image acquisition and preprocessing to cell segmentation and biomarker quantification. We discuss the common open-source tools that support each step to guide users in selecting appropriate solutions. By outlining an end-to-end pipeline with concrete examples, this review is intended for practicing pathologists and researchers with limited computational expertise. It provides practical guidance and best practices to help integrate multiplex image analysis into routine pathology workflows and translational research, bridging the gap between advanced imaging technology and day-to-day diagnostic practice.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104220"},"PeriodicalIF":4.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760448","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Pretreatment with valproic acid alleviates pulmonary fibrosis through epithelial–mesenchymal transition inhibition in vitro and in vivo” [Laboratory Investigation 101 (2021) 1166-1175]","authors":"Lin Chen ,&nbsp;Azeem Alam ,&nbsp;Aurelie Pac-Soo ,&nbsp;Qian Chen ,&nbsp;You Shang ,&nbsp;Hailin Zhao ,&nbsp;Shanglong Yao ,&nbsp;Daqing Ma","doi":"10.1016/j.labinv.2025.104229","DOIUrl":"10.1016/j.labinv.2025.104229","url":null,"abstract":"","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104229"},"PeriodicalIF":4.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Corrigendum to “Histopathological Insights into Metastatic Breast Cancer Gained From Rapid Autopsies” [Laboratory Investigation 105(2025) 104202]","authors":"Gitte Zels ,&nbsp;Anirudh Pabba ,&nbsp;Maxim De Schepper ,&nbsp;Tatjana Geukens ,&nbsp;Karen Van Baelen ,&nbsp;Marion Maetens ,&nbsp;Amena Mahdami ,&nbsp;Sophia Leduc ,&nbsp;Edoardo Isnaldi ,&nbsp;Ha-Linh Nguyen ,&nbsp;Imane Bachir ,&nbsp;Josephine Van Cauwenberge ,&nbsp;Kristien Borremans ,&nbsp;Birgit Weynand ,&nbsp;Elia Biganzoli ,&nbsp;Patrick Neven ,&nbsp;Hans Wildiers ,&nbsp;Wouter Van Den Bogaert ,&nbsp;François Richard ,&nbsp;Christine Desmedt ,&nbsp;Giuseppe Floris","doi":"10.1016/j.labinv.2025.104224","DOIUrl":"10.1016/j.labinv.2025.104224","url":null,"abstract":"","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 10","pages":"Article 104224"},"PeriodicalIF":4.2,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144766414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}