Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

MICROELECTRONIC PHOTOSENSORS MICROELECTRONIC PHOTOSENSORS

FOR GENETIC DIAGNOSTIC MICROSYSTEMSFOR GENETIC DIAGNOSTIC MICROSYSTEMS

MICROELECTRONIC PHOTOSENSORS MICROELECTRONIC PHOTOSENSORS

FOR GENETIC DIAGNOSTIC MICROSYSTEMSFOR GENETIC DIAGNOSTIC MICROSYSTEMS

GE 2003

I. Rech; S. Cova; F. Zappa; M. GhioniPolitecnico di Milano - DEI, piazza. Leonardo da Vinci 32, 20133 Milano, Italy

M. Chiari; M. Cretich

CNR - ICRM, via M. Bianco, 20133 Milano

Ivan RechPolitecnico 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

Ivan RechPolitecnico 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-upRequirements :

Small volumes of the reagents

Ivan RechPolitecnico 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

Ivan RechPolitecnico 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

Avalanche PhotoDiode Single-Photon Avalanche Diode

APD SPAD

Ivan RechPolitecnico 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

n

p++p

p

hv

+n

anode cathode

Planar SPAD structure

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

High QE (Quantum detection Efficiency)

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

0

5

10

15

20

25

30

35

40

0 1 2 3 4 5 6 7 8 9 10 11 12 13

Sovratensione (V)

Cont

eggi

di B

uio

(cou

nts/

sec.

)

100

150

200

250

300

350

400

0 1 2 3 4 5 6 7 8 9 10 11 12 13

Sovratensione (V)Co

nteg

gi d

i Bui

o (c

ount

s/se

c.)

Active area diameter : 10 Active area diameter : 10 mm Active area diameter : 20 Active area diameter : 20 mm

Low internal noise : Dark Counts

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Dark Counts Vs Temperature

0

50

100

150

200

250

300

350

-30 -20 -10 0 10 20

Temperature [C°]

Da

rk C

ou

nts

[c

/s]

+5V

+10V

0

50

100

150

200

250

300

350

-30 -20 -10 0 10 20

Temperature [C°]

Da

rk C

ou

nts

[c

/s]

+5V

+10V

Experimental set-up

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Fragment separation

Electropherogram

Ivan RechPolitecnico 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• Fmax 6.5 MHz• Waveform storage

Politecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

SeparationInjection

Sample

Buffer

Waste

Chip Electrophoresis

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

First Experimental Setup

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Ivan RechPolitecnico 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)

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Results

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

121416181101121141161181

0

10000

20000

30000

40000

50000

60000

70000

80000

Static measure5ng/ul

0,5ng/ul

0,05ng/ul

100pM oligonucleotide labelled with CY 5

300

400

500

600

700

800

900

1000

1100

1200

1300

0 50 100 150 200Time [Sec.]

Co

un

ts [

c/s]

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 limitDetection limit ( with S/N=3) :( with S/N=3) : 3pM3pM (with injection volume 50pL)

Ivan RechPolitecnico 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.XieHarvard Univ. Dept. Chemistry and Chemical Biology

FWHM=60ps

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Prospect

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

• Improved optics sensitivity enhancement to 100fM

• Wider-area SPAD simpler set-up

• monolithic SPAD&AQC micro-miniaturized detection head

• SPAD arrays elaborate detection head

Prospect

Ivan RechPolitecnico di Milano

MICROELECTRONIC PHOTOSENSORS FOR GENETIC DIAGNOSTIC MICROSYSTEMS GE 2003 , June 13, 2003

Acknowledgments

Politecnico di Milano, Italy

Alessandro Restelli Electronics & 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

Ivan RechPolitecnico 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