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