1. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 I. Rech; S. Cova; F. Zappa; M. Ghioni Politecnico di Milano - DEI, piazza. Leonardo da Vinci 32, 20133 Milano, Italy M. Chiari; M. Cretich CNR - ICRM, via M. Bianco, 20133 Milano

2. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Outline Analytical Microsystems and Fluorescence Detection SPAD detectors Experimental set-up Results and prospect

3. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Goal : reduced measurement operation costs higher performance lower cost higher flexibility better automation and control Genetical analysis Analisys set-up Requirements : Small volumes of the reagents

4. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Requirements to Detector high QE (Quantum detection Efficiency) Low internal noise High sensitivity down to Single-Photon detection PMT, Photomultiplier Tubes CCD, Charge -Coupled Devices APD, Avalanche PhotoDiodes SPAD, Single-Photon Avalanche Diodes High-Sensitivity Detectors

5. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Bias: well ABOVE breakdown Geiger-mode: it’s a TRIGGER device!! Gain: meaningless... or “infinite” !! Bias: slightly BELOW breakdown Linear-mode: it’s an AMPLIFIER Gain: limited < 1000 A valanche P hoto D iode S ingle- P hoton A valanche D iode APD SPAD

6. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Well defined high-field region No edge effects No microplasmas Low generation rate Minimum afterpulsing effect Low series resistance Low cost process Low Power dissipation Planar SPAD structure

7. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 High QE (Quantum detection Efficiency)

8. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Active area diameter : 10  m Active area diameter : 20  m Low internal noise : Dark Counts

9. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Dark Counts Vs Temperature

10. Experimental set-up

11. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Fragment separation Electropherogram

12. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Counting Module Single power supply +5V Software controlled settings RS-232 data transmission F max 6.5 MHz Waveform storage Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003

13. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Separation Injection Sample Buffer Waste Chip Electrophoresis

14. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 First Experimental Setup

15. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003

16. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Current Apparatus : SV3 Compact and low-cost Full automated Laser diode for fluorescence excitation Controlled chip temperature Dual wavelength electropherogram Dual HV power supply (0-5kV) Confocal optical scheme Remote control and problem debug (via internet)

17. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003

18. Results

19. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 S/N=35 Glass microchips purchased from Micralyne Inc. (Edmonton, Canada). Electrosmotic flow in the separation channels suppressed by EPDMA dynamic coating. Separation run buffer: TAPS-TRIS 100mM pH 8.5. Sample :23 mer oligonucleotide labelled with CY 5 Detection limit( with S/N=3) : 3pM Detection limit ( with S/N=3) : 3pM (with injection volume 50pL)

20. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 SPTM- Single Photon Timing Module Single power supply +15V High Time-resolution (60ps) Low dark Counts (down to 5 c/sec) Controlled Temperature Software controlled settings RS-232 data transmission Politecnico di Milano Single Molecole Detection Collaboration with S.Xie Harvard Univ. Dept. Chemistry and Chemical Biology FWHM=60ps MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003

21. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Prospect

22. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Improved optics  sensitivity enhancement to 100fM Wider-areaSPAD  simpler set-up monolithic SPAD&AQC  micro-miniaturized detection head SPAD arrays  elaborate detection head Prospect

23. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Acknowledgments Politecnico di Milano, Italy Alessandro RestelliElectronics & Optics Ivan Labanca Dario Vagni Giuseppe Liaci Fabrizio Mingozzi ICRM-CNR, Milano, Italy: Marcella CHIARI Polymers & Electrophoresis Marina CRETICH CEQSE-CNR, Milano, Italy: Luciano PALLARO Mechanics & Optics

24. Ivan Rech Politecnico di Milano MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003, June 13, 2003 Thin SPAD’s Thick SPAD’s 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 IC integrated circuits) Robust and rugged Low-cost NO COMMERCIAL SOURCE TODAY Very good QE and low noise Sub-nanosecond timing High voltage : 300 to 400V High dissipation : Peltier cooler required Ultra-pure high-resistivity Si substrate Dedicated fabrication process (NOT compatible with IC’s) Delicate and degradable Expensive SINGLE COMMERCIAL SOURCE