The Feulgen reaction: from pink-magenta to rainbow fluorescent at the Maffo Vialli’s School of Histochemistry

Abstract

For over a century, Palazzo Botta (Palace Botta) has housed the University of Pavia's Biomedical Institutes. Illustrious scientists have conducted research and taught at this Palace, making significant contributions to the advancement of natural, biological, and medical science. Among them, Camillo Golgi received the Nobel Prize for discovering the so-called "black reaction." Following Golgi, the Palace continued to be a hub for the development of methodologies and reactions aimed at detecting and quantifying biological components. Maffo Vialli (in the Golgi stream) was the first to establish a Histochemistry Research Group, which began in the naturalistic field and later expanded to the biomedical area. Among the many histochemical studies initiated in the Palace, the Feulgen reaction undoubtedly played a significant role. This reaction, developed R. Feulgen and H. Rossenbeck in 1924, had significant international implications: numerous researchers then contributed to define its fine chemical details, which remained the subject of study for years, resulting in a massive international scientific literature. The Pavia School of Histochemistry also contributed to the evolution and application of this method, which has become a true benchmark in quantitative histochemistry. Giovanni Prenna and the CNR Centre for Histochemistry made significant contributions, as they were already focused on fluorescence cytochemistry. The Pavia researchers made significant contributions to the development of methodology and, in particular, instrumentation; the evolution of the latter resulted in the emergence of flow cytometry and an ever-increasing family of fluorescent probes, which somewhat overshadowed the Feulgen reaction for DNA quantification. The advent of monoclonal antibodies then contributed to the final explosion of flow cytometry in clinical application, almost making young neophytes forget that its roots date back to Feulgen.

A brief history of the Feulgen reaction

“…A significant improvement was the development of novel, more stable, and easily reproducible Schiff-type reagents, aiming to overcome limitations such as the intrinsic difficulties inherent in completing the laborious synthesis steps. These innovations allowed the Feulgen reaction to efficiently expand its histochemical applications into areas such as flow cytometry, where it served for quantitative DNA determination for both research and clinical purposes, and in particular electron microscopy (EM), granting DNA visualization with ultrastructural resolution (Casali et al 2022 ; Mazzini 2024 ).…”