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Advanced Photon-Timing Detector Module for Single-Molecule Spectroscopy

This paper presents the development of a novel Photon-Timing Detector Module (SPTM2) for single-molecule spectroscopy, achieving unprecedented temporal resolution below 100 ps. The capabilities of the SPTM2 allow for real-time monitoring of single-molecule dynamics, enabling detailed studies of molecular properties and interactions, such as fluorescence resonance energy transfer (FRET) and structural dynamics. Our experimental results demonstrate significant improvements in quantum efficiency, low noise levels, and compatibility with various applications in biological research.

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Advanced Photon-Timing Detector Module for Single-Molecule Spectroscopy

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  1. POLITECNICO DI MILANO PHOTON-TIMING DETECTOR MODULE FOR SINGLE-MOLECULE SPECTROSCOPY I.Rech*, M. Ghioni*, S. Cova*, H. Yang°, G. Luo°, X.S. Xie° * Dip. di Elettronica e Informazione and IFN-CNR, POLITECNICO DI MILANO ° Dep. of Chemistry and Chemical Biology, HARVARD UNIVERSITY

  2. Sommario : • Introduzione : perché misure su singole molecole • Misure di fluorescenza “time-resolved” • Single Photon Avalanche Diode (SPAD) • Modulo SPTM2 • Risultati sperimentali

  3. Perché misure su singole molecole? • Misure di proprietà di molecole NON MEDIATE SU INSIEME • Possibilità di seguire in tempo reale l’evoluzione • Informazioni sulla dinamica della molecola Molecole di interesse biologico : Proteine FRET : Fluorescence Resonance Energy Trasnfer Variazioni di struttura nanometriche

  4. Single Molecule Structural Dynamics

  5. Nuovo metodo di X.S. Xie, Harvard University - Basato su eccitazione con impulsi laser - Rivelazione fotoni fluorescenti e misure di: • Ritardo emissione risp. eccitazione • Tempo reale di emissione • Analisi per correlazione Excitation pulse train t1 t2 t3 photon

  6. Setup di Misura ?

  7. Requisiti del Rivelatore • Alta QE (Quantum detection Efficiency) > 40% • Basso rumore intrinseco < 50c/s • Alta Risoluzione Temporale (FWHM) < 100ps NON ESISTONO RIVELATORI COMMERCIALI CON QUESTE CARATTERISTICHE

  8. Single Photon Avalanche Diode (SPAD)

  9. PerkinElmer SPCM (SLIKTM diode) • QE: 30% (@500nm) OK • Dark Counts: 150 c/s OK • FWHM: >300ps NO! • High voltage : 300 to 400V • High dissipation • Delicate and degradable • Very expensive Reach-Trough structure Typical active region: 200 m diameter

  10. Sviluppo SPAD (Polimi e IMM-CNR) • Good QE and low noise • Picosecond timing • Low voltage : 15 to 40V • Low power : cooling not necessary • Standard Si substrate • Planar fabrication process • COMPATIBLE with array detector and integrated circuits • Robust and rugged • Low-cost Planar structure typical active region: 20-50 m diameter

  11. Quantum Efficiency e Dark Counting Rate

  12. Modulo SPTM2

  13. Modulo SPTM2 • Compatto (82x60x30mm) • Singola alimentazione • Tensioni programmabili • Hold off programmabile • Interfaccia RS-232 • Controllo di Temperatura • Pulse Counter integrato Sfruttare le Performance del Rivelatore Sistema Compatto e User Friendly

  14. APD SPAD Avalanche PhotoDiode APD Linear-mode: AMPLIFICATORE Single-Photon Avalanche Diode SPAD Geiger-mode: TRIGGER device!!

  15. iAQC: integrated Active Quenching Circuit • Active Quenching Circuit • Tempo morto corto e ben definito • High counting rate > 1 Mc/s • Impulso Standard d’uscita • iAQC, oltre la miniaturizzazione… • Capacità ridotte • Carica di valanga ridotta • Afterpulsing ridotto F.Zappa, S.Cova, M.Ghioni, US patent

  16. Modulo SPTM2: Diagramma a Blocchi

  17. Risoluzione Temporale PerkinElmer SPCM (SLIKTM diode) Planar thin Si-SPAD

  18. Risoluzione Temporale @500nm

  19. Modulo SPTM2

  20. Risultati Sperimentali

  21. Misure su Singola Molecola

  22. “Single-Molecule Protein Dynamics on Multiple Time Scales Probed by Electron Transfer” Haw Yang*, Guobin Luo*, Pallop Karnchanaphanurach*, Tai-Man Louie‡, Ivan Rech°, Sergio Cova°, Luying Xun‡ and X. Sunney Xie* * Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138. ‡ School of Molecular Biosciences, Washington State University, Pullman, WA 99164. ° Dipartimento di Elettronica e Informazione, Politecnico di Milano Science, Vol.302, 262-266,10 Oct. 2003. “Photon-Timing Detector Module for Single-Molecule Spectroscopy with 60ps Resolution” Ivan Rech, Guobin Luo*, Masimo Ghioni, Sergio Cova, Haw Yang*, X. Sunney Xie* Dipartimento di Elettronica e Informazione, Politecnico di Milano *Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA IEEE Journal of Selected Topics in Quantum Electronics, in press, 2004

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