The Seminal Contributions of Gregorio Weber to Modern Fluorescence Spectroscopy

Abstract

Gregorio Weber is acknowledged to be the person responsible for many of the more important theoretical and experimental developments in modern fluorescence spectroscopy. In particular, Weber pioneered the application of fluorescence spectroscopy to the biological sciences. His list of achievements includes:-The synthesis and use of dansyl chloride as a probe of protein hydrodynamics -The extension of Perrin's theory of fluorescence polarization to fluorophores associated with random orientations with ellipsoids of revolution and to mixtures of fluorophores -The first spectral resolution of the fluorescence of the aromatic amino acids and of intrinsic fluorescence of proteins -The first demonstration that both FAD and NADH make internal complexes -The first report on aromatic secondary amines, which are strongly fluorescent in apolar solvents, but hardly in water, the most spectacular case being the anilino-naphthalene sulfonates CANS) -The first description of the use of the fluorescence of small molecules as probes for the viscosity of micelles, with implications for membrane systems -A general formulation of depolarization by energy transfer -The discovery of the "red-edge" effect in homo-energy transfer -The development of modern cross-correlation phase fluorometry -The development of the excitation-emission matrix method for resolving contributions from multiple fluorophores -The synthesis of several novel fluorophores, including pyrenebutyric acid, IAEDANS, bis-ANS, PRODAN, and LAURDAN, designed to probe dynamic aspects of biomolecules In addition to these seminal contributions, Gregorio Weber also trained and inspired generations of spectroscopists and biophysicists who went on to make important contributions in their fields, including both basic research as well as the commercialization of fluorescence methodologies and their extension into the clinical and biomedical disciplines. OverviewDuring the last few decades, fluorescence spectroscopy has evolved from a narrow, highly specialized technique into an important discipline widely utilized in the biological, chemical, and physical sciences. Fluorescence methodologies have also assumed an increasingly important role in the clinical and medical sciences. There are now world-renowned centers for fluorescence spectroscopy, highly successful commercial enterprises specializing in fluorescence instrumentation, fluorescence based clinical instruments in virtually all hospital laboratories, and thousands of practitioners worldwide. As in all scientific disciplines, the development of modern fluorescence spectroscopy has benefited

The evolution of spectral flow cytometry

“…The roots of spectral flow cytometry, as for many things fluorescence, can be traced to the work of Gregorio Weber [2], who pioneered the use of fluorescence to study biological systems. Weber, working at the Universities of Sheffield and Illinois, established the conceptual and practical underpinnings of bioanalytical fluorescence spectroscopy, integrating chemistry, physics, and engineering to produce dyes, instruments, and experimental approaches to study systems ranging from proteins to cells [3]. At about the time Fulwyler was building the first single-cell sorter [4], and some years before the first fluorescence flow cytometers [5][6][7], Weber, recognizing the wealth of information contained in the excitation and emission spectra of fluorescent compounds, described the determination of the number and abundances of multiple fluorescent species in a mixed sample using the excitationfluorescence (EF) matrix [8].…”

Overlapping synthetic peptides as a tool to map protein-protein interactions ̶ FSH as a model system of nonadditive interactions

“…Remembering a series of lectures given by Gregorio Weber during one of his visits to Buenos Aires in 1986, and his previous work [112][113][114][115] , I decided to test whether the individual binding regions of FSH-β behaved additively. Before, William P. Jencks had described the principle of ΔG additivity and cooperativity applied to proteins 116 , based on Weber's ΔG of coupling 112 , or the earlier and equivalent Wyman's ΔG of interactions 117 .…”