FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING

In the last 43 years, thanks to fantastic collaborators, we used antibodies in immunostaining and published a few firsts: TP53 in breast cancer (1988), FFPE-proofs anti Ki-67 antibody MIB 1 and the citrate antigen retrieval buffer (1992), reticular cells in human bone marrow (1993), BCL6 in germina...

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Language:English
Published: PAGEPress Publications 2025-08-01
Series:European Journal of Histochemistry
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Online Access:https://www.ejh.it/ejh/article/view/4279
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description In the last 43 years, thanks to fantastic collaborators, we used antibodies in immunostaining and published a few firsts: TP53 in breast cancer (1988), FFPE-proofs anti Ki-67 antibody MIB 1 and the citrate antigen retrieval buffer (1992), reticular cells in human bone marrow (1993), BCL6 in germinal centers (1995), PRDM1 in plasma cells (2005), the sub cellular localization and distribution of AID and the function of IRF4 (2006). From 2013 to 2021 we dissected the effects of tissue processing on antigenicity, a body of discoveries which led to a hyperplexed (>15 markers) multiplexing method, Multiple Iterative Labeling by Antibody Neodeposition (MILAN)1,2. The application of MILAN and the complexity of data obtained (~100 markers, millions of single cells) landed us in the rarefied world of dimensionality reduction algorithms, where math rules and statistical significance replaces “representative images”. Because of these latest developments, we discovered that the human eye ability to discriminate shades of grey is very limited (less than 64/256)3, thus low levels of staining are routinely missed and eye-guided assessment is unreliable. Bioinformatics tools we developed (BRAQUE)4 provides highest sensitivity, granularity and robustness based on objective statistical parameters. Biologists and Pathologists need to believe (in math)5 rather than see with their own flawed eyes.
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spelling doaj-art-5f31774a4ff6426cb881e9ae3782e2bd2025-08-23T11:19:53ZengPAGEPress PublicationsEuropean Journal of Histochemistry1121-760X2038-83062025-08-0169s210.4081/ejh.2025.4279FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING In the last 43 years, thanks to fantastic collaborators, we used antibodies in immunostaining and published a few firsts: TP53 in breast cancer (1988), FFPE-proofs anti Ki-67 antibody MIB 1 and the citrate antigen retrieval buffer (1992), reticular cells in human bone marrow (1993), BCL6 in germinal centers (1995), PRDM1 in plasma cells (2005), the sub cellular localization and distribution of AID and the function of IRF4 (2006). From 2013 to 2021 we dissected the effects of tissue processing on antigenicity, a body of discoveries which led to a hyperplexed (>15 markers) multiplexing method, Multiple Iterative Labeling by Antibody Neodeposition (MILAN)1,2. The application of MILAN and the complexity of data obtained (~100 markers, millions of single cells) landed us in the rarefied world of dimensionality reduction algorithms, where math rules and statistical significance replaces “representative images”. Because of these latest developments, we discovered that the human eye ability to discriminate shades of grey is very limited (less than 64/256)3, thus low levels of staining are routinely missed and eye-guided assessment is unreliable. Bioinformatics tools we developed (BRAQUE)4 provides highest sensitivity, granularity and robustness based on objective statistical parameters. Biologists and Pathologists need to believe (in math)5 rather than see with their own flawed eyes. https://www.ejh.it/ejh/article/view/4279DEVELOPMENT, VALIDATION AND APPLICATION OF MULTIPLEX IMMUNOHISTOCHEMICAL ASSAYS
spellingShingle FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
European Journal of Histochemistry
DEVELOPMENT, VALIDATION AND APPLICATION OF MULTIPLEX IMMUNOHISTOCHEMICAL ASSAYS
title FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
title_full FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
title_fullStr FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
title_full_unstemmed FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
title_short FROM SEEING TO BELIEVING: A 43-YEAR LONG JOURNEY IN IMMUNOSTAINING
title_sort from seeing to believing a 43 year long journey in immunostaining
topic DEVELOPMENT, VALIDATION AND APPLICATION OF MULTIPLEX IMMUNOHISTOCHEMICAL ASSAYS
url https://www.ejh.it/ejh/article/view/4279