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      Single Photon Avalanche Diode Arrays for Time-Resolved Raman Spectroscopy

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          Abstract

          The detection of peaks shifts in Raman spectroscopy enables a fingerprint reconstruction to discriminate among molecules with neither labelling nor sample preparation. Time-resolved Raman spectroscopy is an effective technique to reject the strong fluorescence background that profits from the time scale difference in the two responses: Raman photons are scattered almost instantaneously while fluorescence shows a nanoseconds time constant decay. The combination of short laser pulses with time-gated detectors enables the collection of only those photons synchronous with the pulse, thus rejecting fluorescent ones. This review addresses time-gating issues from the sensor standpoint and identifies single photon avalanche diode (SPAD) arrays as the most suitable single-photon detectors to be rapidly and precisely time-gated without bulky, complex, or expensive setups. At first, we discuss the requirements for ideal Raman SPAD arrays, particularly focusing on the design guidelines for optimized on-chip processing electronics. Then we present some existing SPAD-based architectures, featuring specific operation modes which can be usefully exploited for Raman spectroscopy. Finally, we highlight key aspects for future ultrafast Raman platforms and highly integrated sensors capable of undistorted identification of Raman peaks across many pixels.

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          Fluorescence lifetime imaging--techniques and applications.

          W. Becker (2012)
          Fluorescence lifetime imaging (FLIM) uses the fact that the fluorescence lifetime of a fluorophore depends on its molecular environment but not on its concentration. Molecular effects in a sample can therefore be investigated independently of the variable, and usually unknown concentration of the fluorophore. There is a variety of technical solutions of lifetime imaging in microscopy. The technical part of this paper focuses on time-domain FLIM by multidimensional time-correlated single photon counting, time-domain FLIM by gated image intensifiers, frequency-domain FLIM by gain-modulated image intensifiers, and frequency-domain FLIM by gain-modulated photomultipliers. The application part describes the most frequent FLIM applications: Measurement of molecular environment parameters, protein-interaction measurements by Förster resonance energy transfer (FRET), and measurements of the metabolic state of cells and tissue via their autofluorescence. Measurements of local environment parameters are based on lifetime changes induced by fluorescence quenching or conformation changes of the fluorophores. The advantage over intensity-based measurements is that no special ratiometric fluorophores are needed. Therefore, a much wider selection of fluorescence markers can be used, and a wider range of cell parameters is accessible. FLIM-FRET measures the change in the decay function of the FRET donor on interaction with an acceptor. FLIM-based FRET measurement does not have to cope with problems like donor bleedthrough or directly excited acceptor fluorescence. This relaxes the requirements to the absorption and emission spectra of the donors and acceptors used. Moreover, FLIM-FRET measurements are able to distinguish interacting and noninteracting fractions of the donor, and thus obtain independent information about distances and interacting and noninteracting protein fractions. This is information not accessible by steady-state FRET techniques. Autofluorescence FLIM exploits changes in the decay parameters of endogenous fluorophores with the metabolic state of the cells or the tissue. By resolving changes in the binding, conformation, and composition of biologically relevant compounds FLIM delivers information not accessible by steady-state fluorescence techniques. © 2012 The Author Journal of Microscopy © 2012 Royal Microscopical Society.
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            A New Type of Secondary Radiation

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              Vibrational and Rotational Relaxation

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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Sensors (Basel)
                Sensors (Basel)
                sensors
                Sensors (Basel, Switzerland)
                MDPI
                1424-8220
                23 June 2021
                July 2021
                : 21
                : 13
                : 4287
                Affiliations
                Dipartimento di Elettronica, Informazione e Bioingegneria, Politecnico di Milano, Via G. Ponzio 34/5, 20133 Milano, Italy; federica.villa@ 123456polimi.it
                Author notes
                Author information
                https://orcid.org/0000-0001-8384-2569
                https://orcid.org/0000-0002-9840-0269
                Article
                sensors-21-04287
                10.3390/s21134287
                8272195
                34201576
                4140a5d2-c445-46ba-bfb7-6f71b3e659fc
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 13 May 2021
                : 18 June 2021
                Categories
                Review

                Biomedical engineering
                time-resolved raman spectroscopy,fluorescence suppression,single photon avalanche diode (spad),spad array,time gating,single photon counting (spc),time-correlated single photon counting (tcspc)

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