Light Induced Fluorescence Spectroscopy Imaging Diagnosis of

Precancerous Lesions

螢光光譜影像技術應用於早期螢光光譜影像技術應用於早期癌組織診斷癌組織診斷

江惠華 陽明大學 醫學工程研究所

Physiological Imaging

• X-rays, CT• Ultrasound• PET• MRI• Near Infrared Light Attenuation Imaging• Photoacoustic Imaging• OCT(Optical Coherent Tomography)• LIF (Light Induced Fluorescence Imaging)

Natural Fluorophores in Tissues

Fluo. Peaks: 390 450 530 nm

Schomacker KT,. Lasers Surg Med, 1992; 12: 63-78

Light Induced Fluorescence

GroundState

ExcitedState

hνA

hνA hν

F

Internal Conversion

Intersystem Crossing

MetastableState

FluorescencePhosphrescence

hνP

S0

S1

S2

T1

01

2

9.0

0.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

650.0350.0375.0400.0425.0450.0475.0500.0525.0550.0575.0600.0625.0

371nm

H2O Fluorescence

LIAF History

• 1989: Lung Cancer (Tang, et al.)

• 1990: Colon Adenoma (Cothren, et al.)

• 1992: Colon Tissues Autofluorescence Mechanism

(Schomacker)

• 1994: Cervics CIN (Richards-Kortum)

• 1997: Esophagus Cancer (Vo-Dinh et al.)

• 1996: PCA Spectroscopy Analysis (Ramanujam)

• 1998: PLS Spectroscopy Analysis (Chiang et al.)

• 1999: Logistic Regression Analysis (Chiang et al.)

Cancer Death Increased RateCancer Death Increased Rate

OralColon

Cervical

-8

-5

-2

1

4

7

10

19961996--97 97 (NIH, ROC)(NIH, ROC)

GI Tract

EEM (Excitation-Emission Matrix )

光譜實驗架構

Excitation / LIAF Emission

Ex/Em ~ 200 X

Typical Autofluorescence Spectra

0

0.5

1

1.5

2

2.5

3

340373406439472505538571604nm

Normalized intensity

UntreatedW2 W4W6 W8 W10W12 W14 W16

Excitation at 320 nm

EEM: Fluorescence excitation-emission matrices (Colon tissue)

PLS Analysis0.15

-0.20

-0.15

-0.10

-0.05

0.00

0.05

0.10

650350380400420440460480500520540560580600620

P1

P2

0.14

0.00

0.02

0.04

0.06

0.08

0.10

0.12

650350380400420440460480500520540560580600620

Normal

Polyp

Cancer P1

P2

0.14

0.00

0.02

0.04

0.06

0.08

0.10

0.12

650350 380400420440460480500520540560580600620

normal

polyp

cancer

Colorectal Autofluorescence Spectra

0

20

40

60

80

Collagen NADH FAD Hemoglobin

Com

pone

nt (%

)

Normal Colonic TissuePoly Colonic TissueCancer Colonic Tissue

Cervical Cancer Screening

臨床試用準確率95%,偽陰性偽陽性都低

UV光/陰道鏡立即分析

UV自體螢光檢查

費用優缺點採檢方法

自費1000~1500準確率95%境外執行疑慮

檢體腦抹片香港分析1~2週

雷射抹片

自費 800準確率90%偽陽性高適合年輕女性

傳統抹片/醋酸螢光目視檢查

螢光抹片(Papsure)

健保準確率60%~85%高偽陰性

抹片刷取子宮頸表皮,約1週可知結果Pap smear

0.8

0.690.7

0.77

0.6

0.65

0.7

0.75

0.8

0.85

0.9

ColposcopyPap smearHPV testingFluorescence

(M. Feld, MIT, Ob. Gyne., vol. 93, no. 3, 1999)

Sensitivity = Specificity Q Point of cervical samples

EndoCerix autofluorescence

(Endo)Cervix Autofluorescence Spectrum

0

0.001

0.002

0.003

0.004

0.005

0.006

370

378

386

394

402

410

418

426

434

442

450

458

466

474

482

490

498

506

514

522

530

538

546

554

562

570

Wavelength (nm)In

tens

ity (

unit)

Normal

Chronic cervicitisCervical SCC

(Endo)Cervix Autofluorescence Spectrum(Normalization)

0

0.002

0.004

0.006

0.008

0.01

0.012

370

378

386

394

402

410

418

426

434

442

450

458

466

474

482

490

498

506

514

522

530

538

546

554

562

570

Wavelength (nm)

Rel

ativ

e In

tens

ity

NormalChronic cervicitisCervical SCC

Collagen380nm

NADH 460nm

Collagen :

Normal > Cancer

NADH :Normal < Cancer

Hb 420nm

Cervical cancer spending

• Up to 50% false positives of colposcopies. • Much caused by inaccurate screening.

MediSpectra's Detection Technology

• Founded in 1996, Lexington, MA• Combine LIF and white light backscatter

spectroscopy• Cervical normal, CIN I, and CIN II/III tissue• FDA 510K

SpectraSCIENCEOptical Biopsy System

• WavSTAT II, FDA approved, 2001.• Minneapolis, MN• Esophageal cancer, colon, application• Performed at Mayo Clinic, MGH

LIF Imaging Participators

• LIFE-Lung (Xillix Technologies Corp; Richmond, BC, Canada) • Xilix received FDA 510K, 1996; • FDA nod for cancer detection device test - 2003-07-14

• D-Light/AF system (Karl Storz Imaging; Tuttlingen, Germany)Leonhard M. New incoherent autofluorescence/ fluorescence system for early

detection of lung cancer. Diagn Ther Endosc 1999; 5:71-75

• SAFE-1000 (Pentax; Tokyo, Japan)Adachi R, Development of the autofluorescence endoscope imaging system. Diagn Ther Endosc 1999; 5:65-70

1. LIAF Spectra and Images

2. Endoscope integration

3. Real time/operator independent

diagnosis

4. Biopsy location selection

Future Developments of LIAF Future Developments of LIAF Tissues DiagnosisTissues Diagnosis

0.14

0.00

0.02

0.04

0.06

0.08

0.10

0.12

650350380400420440460480500520540560580600620

Normal

Polyp

Cancer