UNIVE D RS N IT U Y L MEDICAL Lund University Medical Centre, Sweden

LASER CENTRE 1991

SWEDEN Challenges in Biophotonics

Therapeutic and Diagnostic applications of in

Laser

• Laser and ablation • Thermo therapy and photocoagulation therapy • Tatoo removal • Cosmetic hair removal •

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

1 Why are used for medical treatments?

– Effective local treatment – Minimally invasive – Minimal side-effects – No bleeding – Non-contact

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Log10 absorption coefficient

H2 O Adenine HbO2 DOPA-Melanin

H O 2 Photofrin

100 200 300 500 700 1000 2000 3000 5000 10,000

UNIVER D SI Wavelength (nm) N T U Y L MEDICAL Ar- LASERUV CENTRE Lund University Medical Laser Centre,PDT Sweden IR 1991 ion SWEDEN Pulsed CO ArF XeCl dye Nd:YAG 2

2 Medical Laser Therapy

• Laser surgery – Eye (Ar-ion , Nd:YAG, Excimer lasers)

– Dermatological (CO2, Dye, Ruby, Ar-ion lasers)

– General surgery (Nd:YAG, diode, CO2 lasers) • Thermo therapy • Photodynamic therapy

UNIVE D RS N IT U Y L Dosimetry is essential!! MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

LASIK Laser-Assited In Situ Keratomileusis

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

3 LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

1. Mark the cornea with inc. Apply anaesthetic eye drops!

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

2a. Fixate the eye with a suction ring (Option: 2b. Wavefront analysis of the eye) UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

4 LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

3. Peel a slice of the cornea with a microkeratome

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

4. Expose the underlying layer of the cornea.

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

5 LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

5. Laser alignment of the eye.

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

Typical values

Wpulse= 3mJ, tpulse= 18 ns λ=193 nm f = 50Hz 6. Reshape the cornea! (Excimer laser) ∅beam= 0.9 mm

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

6 LASIK Laser-Assited In Situ Keratomileusis Seven steps towards a perfect sight

7. Put the flap back, wait 5 minutes, then go home!

UNIVE D RS N IT U Y L MEDICAL From www.eyesurgeryeducation.com

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Medical Laser Therapy

• Laser surgery – Eye (Ar-ion , Nd:YAG, Excimer lasers)

– Dermatological (CO2, Dye, Ruby, Ar-ion lasers)

– General surgery (Nd:YAG, diode, CO2 lasers) • Thermo therapy • Photodynamic therapy

UNIVE D RS N IT U Y L Dosimetry is essential!! MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

7 Laser Skin Resurfacing

UNIVE D RS N IT U Y L MEDICAL Wrinkles

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy: Stuart Nelson, UC Irvine

Laser Skin Resurfacing

UNIVE D RS N IT U Y L MEDICAL Rhinophyma

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy: Stuart Nelson, UC Irvine

8 Laser Skin Resurfacing

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy: Stuart Nelson, UC Irvine

Formation of New Collagen

Pre Treatment

UNIVE D RS N IT U Y 6 Months Post Treatment L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy: Stuart Nelson, UC Irvine

9 What is Non-Ablative Skin Rejuvenation? Epidermal protection New collagen deposition

UNIVE D RS N IT U Y L MEDICAL Heating of upper dermis New collagen deposition LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy: Stuart Nelson, UC Irvine

Port Wine Stains

DEEP RED/PURPLE PWS

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy Lars Svaasand, NTNU Trondheim

10 NORMAL SKIN

Stratum corneum EPIDERMIS 0.1 mm Melanosomes Papillary dermis Upper vascular plexus

1 mm DERMIS

Lower vascular plexus

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy Lars Svaasand, NTNU Trondheim

PORT WINE STAIN

EPIDERMIS

Papillary dermis Upper vascular plexus

DERMIS

Lower vascular plexus

UNIVE D RS N IT U Y L MEDICAL Ectactic venules/abnormal density of vessel LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN Courtesy Lars Svaasand, NTNU Trondheim

11 ABSORPTION and SCATTERING SPECTRA

Melanin (arb.units)

Scattering (arb. units)

UV IR UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Medical Laser Therapy

• Laser surgery – Eye (Ar-ion , Nd:YAG, Excimer lasers)

– Dermatological (CO2, Dye, Ruby, Ar-ion lasers)

– General surgery (Nd:YAG, diode, CO2 lasers) • Thermo therapy • Photodynamic therapy

UNIVE D RS N IT U Y L Dosimetry is essential!! MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

12 Laser Medical pen

•Both for cutting tissue and for coagulation of bleedings •Laser characteristics: P = 5W, λ = 980 nm

UNIVE D RS N IT U Y L Developed by the US Air Force MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Laser-Tissue Interaction Mechanisms Power density (W/cm2) 1015 Electro- 1012 mechanics Ophthalmology Ablation 109 Angioplasty 104 J/cm2

106 1 J/cm2 Thermal effects Surgery 103 Tissue welding Hyperthermia 1 Photochemistry PDT Biostimulation

UNIVE D RS N IT -9 -6 -3 3 U Y L 10 10 10 1 10 MEDICAL Pulse duration (s) LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

13 Optical measurements of tissue •Morphological measurements – White light imaging and Microscopy – Optical coherence tomography (OCT) – Optical mammography •Physiological measurements – Oxygenation – Optical biopsy – Transcutaneous glucose measurements – Body fluid constituents (dialysis monitoring, blood vials analysis) •Velocity measurements – Laser Doppler – Breathing UNIVE D RS N IT U Y L MEDICAL – Photoplethysmography (PPG)

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

“OCT for dummies” Optical Coherence Tomography

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

14 Commercial OCT system

Humphrey® Optical Coherence Tomography Scanner

UNIVE D RS N IT U Y at Herlev Hospital. L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 Courtesy Peter Andersen, Risø, Denmark

SWEDEN

Applications: Ophthalmology

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 Courtesy Peter Andersen, Risø, Denmark

SWEDEN

15 Incoming light

Nerve fiber layer Ganglion cell layer Inner plexiform layer

Inner nuclear layer

Outer plexiform layer Outer nuclear layer

External limiting membrane Layers of rods and cones Retinal pigment epithelium Choroid

Sclera UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 Courtesy Peter Andersen, Risø, Denmark

SWEDEN

Opportunities for New Technologies

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

16 Consequences of Age Related Changes

m x-ray mammograms (normal breasts)

t

h

.

1

5

y

t

h

b

8

a

/

h

t

l

a

e

h

/

s

t

p

e

d

/

m

o

c

.

s

t

r

a MILDY

e

m FATTY o DENSE DENSE

h

/

/

:

p

t

t

h

UNIV AGE E D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Mammography: Good and Bad Overall ~90% sensitivity, specificity Post-Menopause Pre-Menopause

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

17 PTB Multi-Channel Scanning Laser Pulse Mammograph

(mark #2, 2003) 652 nm

• 4 (8) detection channels Æ off-axis data (zT) 684 nm • 4 wavelengths ÆcHb, cHbO2 + water + lipid 797 nm Æscatter power 839 nm

now: 4 λ multiplexed

planned:

y+2cm

UNIVE D RS N IT U Y L MEDICAL x-2cm x x+2cm

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Patients #144: Cysts (Mammot3)

Thickness: 4.8 cm (CC) 5.0 cm (OB) Scattering 637 nm 656 nm 683 nm 785 nm

Lesion size: 20 mm

Lesion size: 40 mm

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 SWEDEN Courtesy Alessandro Torrecelli, POLIMI

18 Patient #130: Cyst (Scattering)

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 SWEDEN Courtesy Alessandro Torrecelli, POLIMI

Construction of 3D scanning bed

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 SWEDEN Courtesy Jeremy Hebden, UCL London

19 Evaluation of contrast & spatial resolution

Absorbing / scattering target

165 mm

50 mm Extra section

Hemisphere

UNIVE D RS N IT Scattering fluid U Y L MEDICAL Intralipid + dye LASER CENTRE Lund University Medical Laser Centre, Sweden 1991 SWEDEN Courtesy Jeremy Hebden, UCL London

NETWORK Hand-Held ANALYZER spectrometer Optical Scanner BIAS DC NETWORK

DIODE OPTICAL LASERS SWITCH

CW Source

APD

OPTICAL 2.8 CM FIBER APD

2.2 CM 5 CM

UNIVE D RS N IT U Y L MEDICAL TO LASER CENTRE Lund University Medical Laser Centre, Sweden INSTRUMENT Pham,1991 TH., et al. Review of Scientific Instruments, 71 , 1 – 14, (2000). SWEDEN Courtesy Bruce Tromberg, UC Irvine Bevilacqua, F., et al. Applied , 39, 6498-6507, (2000).

20 Fluorescence? Molecular energy

Absorption Fluorescence of light emission

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Clinical questions ∗ Discriminate adenoumatous and hyperplastic polyps in the colon ∗ Identify premalignant and malignant lesions of the vocal fold ∗ Identify malignant tissue in the brain ∗ Visualize the borders of malignant skin lesions

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

21 Fluorescence diagnostics

• Optical contact fibre or through surgical microscope • Non-invasive • Real-time diagnosis • Provide “optical biopsy” information • Endoscopy UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

• Autofluorescence Collagen Elastin NADH...

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

22 Fluorescence: Tissue autofluorescence Protoporphyrin IX

337 nm excitation 405 nm excitation

Collagen ElastinNADH Carotene Protoporphyrin IX Fluorescence [a.u.] intensity

400 500 600 700 500 550 600 650 700 750 Wavelength (nm) Wavelength (nm)

UNIVE D RS N IT U Y L J. Johansson, Dissertation thesis, LTH (1993). MEDICAL af Klinteberg et al. (1999)

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Clinical measurements

Hyperplastic: Adenomatous:

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

23 Gated Fluorosensor

Synkronisation

Trigger fibre Nitrogen laser

337 nm

Detector

Dye laser (405 nm) Spectrometer

1998- Optics wheel UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden Optical fibre 1991

SWEDEN

Fluorosensors

Nitrogen laser based fluorosensor

UNIVE D RS N IT U Y L MEDICAL Diode laser based fluorosensor LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

24 Diode laser fluorosensor 1999 Diode laser 396 nm

Beam splitter Optical fibre

Spectrometer

2001 Shutter Software

Lap top controlled

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Old, old systems

1986 1990

1999 2003 R.I.P. R.I.P.

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

25 Fluorescence larynx

Ventricular fold anterior Vocal fold

~5 mm

• 337 nm excitation • Multivariate methods for classification: SIMCA, PCA, PLS, logistic regression. • Malignant and premalignant lesions vs. normal tissue UNIVE D RS N IT U Y L ⇒ up to 88% sens. and 93% spec. MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Autofluorescence spectra from tissue

337 nm excitation

Normal

Intensity [a.u.] Pre-malignant

400 450 500 550 600 650 Emission wavelength [nm]

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

26 • Autofluorescence Collagen Elastin NADH...

• Fluorescence induced by tumour marker e.g. ALA-induced PpIX

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Typical fluorescence spectra

Normal Malignant Intensity

400 500 600 700nm UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

27 Exogenous fluorescence and Photodynamic therapy (PDT) 1. Administration of ALA 3. Diagnostics

Haem 635 nm

Iron

ALA Protoporfyrin IX Blue laser

2. Production of PpIX

UNIVE D RS N IT U Y L Tumour MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Fluorescence diagnostics of skin tumour following ALA administration

35 Border

35 Healthy skin 35 Tumour 0 500 600 700

0 0 500 600 700 500 600 700

UNIVE D RS N IT U Y L MEDICAL Basal Cell Carcinoma of the skin

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

28 Fluorescence spectra

1 hour post PDT Post PDT Pre PDT Normal skin 4 hours post ALA 10 mm outside lesion 400 450 500 550 600 650 700 750 2 hours post ALA

Wavelength (nm) 1 hour post ALA Pre ALA

BCC

Centre of lesion Intensity Intensity

UNIVE D RS N IT U Y L MEDICAL 400 450 500 550 600 650 700 750

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Tissue fluorescence with ALA-PpIX as tumour marker

405 nm excitation

Normal Adenoma Intensity [a.u.]

UNIVE 450 500 R 550 600 650 D S 700 N IT U Y L MEDICAL Emission wavelength [nm] LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

29 PpIX fluorescence Autofluorescence

5 mm 5 mm

7000 70000 6000 Astrocytoma 60000 5000

4000 50000

3000 40000

2000 30000 1000 20000 0 440 490 540 590 640 690 -1000 10000

-2000 0 400 450 500 550 600 650 700 15 mm 15 mm 140000 120000 120000 100000 100000 80000 80000 60000 60000 40000 40000 20000 20000 0 0 400 450 500 550 600 650 700 440 490 540 590 640 690

35 mm 35 mm 90000 30000 80000 70000 25000 60000

20000 50000 40000 15000 30000 20000 10000 10000 0 5000 UNIVE D RS 400 450 500 550 600 650 700 N IT U Y L MEDICAL0 440 490 540 590 640 690

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

30 Fluorescence diagnostics

Point monitoring: Whole spectrum in one small tissue site

Imaging: Less spectral information but in larger area

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Fluorescence specrum

Fitted curve PpIX peak A Ratio = A Autofluorescence B Intensity

B

400 450 500 550 600 650 700 750 Wavelength (nm) UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

31 Fluorescence imaging

TAE nBCC

G R Intensified B sBCC CCD camera nBCC

TAE Laser

5 Optical A 4 fiber 3 Beam 2 splitterD White-light 1 Fluorescence Intensity (a.u.) Intensity Fluorescence CCD camera 0 450500550600650700750 Wavelength (nm)

8 7 6 Lens 5 4 3 2 UNIVE D RS N IT 1 U Y L MEDICAL 0

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Multicolour Fluorescence Imaging

White light image

Digitally processed image

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

32 Hyperspectral imaging system

Sagnac interferometer

CCD detector

INTERFEROGRAM collimating lens 0.8 light source 0.6

barrier filter 0.4 dichroic filter 0.2 0.0 excitation filter -0.2

-0.4 objective -0.6

-0.8 UNIVE D RS -10 -5 0 5 10 N IT U Y L MEDICAL Patient Fluorescence intensity Interferometer rotation angle LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Hyperspectral imaging

Fluorescence image using White light photo visual colour scale Fluorescence intensity (a.u.)

400 500 600 700 800 Wavelength (nm)

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

33 Hyperspectral fluorescence imaging of a Basal Cell Carcinoma

Pre ALA Pre PDT Post PDT

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Hyperspectral fluorescence imaging of a BCC pre PDT

Photo Visual colour coding

Similarity plot PLS analysis

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

34 Hyper Spectral Diagnostic Imager

Combines: • video image • reflectance scan • fluorescence scan

Aim: • interactive diagnostics • integrated colposcope

UNIVE D RS N IT U Y L MEDICAL Under development by Science & Technology Inc, USA

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Fibre endoscope

•Coherent fibre bundle

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

35 Cross-section of the distal end

Air/fluid channel

Image channel Guided light fibre bundle

Biopsy channel

Guided light fibre bundle

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

The diagnosis

White-light mode Blue-light mode

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

36 Measurements at Ear-Nose- Throat clinic

Larynx

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Fluorescence diagnostics of skin tumour following ALA administration 1. Administration of ALA 3. Diagnostics

Haem 635 nm

Iron

ALA Protoporfyrin IX Blue laser

2. Production of PpIX 4. Treatment

Red laser

UNIVE D RS N IT U Y L Tumour MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

37 Energy level diagram for PDT

Hp O2

S2 1 + T2 B Σ g

IX S1 T1 1 630 A Δ g nm γ F 1270 P nm

3 - X Σ g S0

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Course of healing

Cryosurgery

Pre 8 days post 7 weeks post 3 months post ALA-PDT

Pre 7 days post 7 weeks post 3 months post

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

38 ALA-PDT of non-melanoma malignant lesions in the region close to the eye

Pre treatment Post treatment

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Parameters of concern in photodynamic therapy

PHOTOSENSITIZER ILLUMINATION Pre-formed agent/porphyrin precursor Wavelength Dose Light dose Mode of administration Fluence rate Tissue localization 1 02 Split-dose regimen Selectivity Photoproduct generation Kinetics

UNIVE D RS N IT U Y L MEDICAL OPTIMIZED TREATMENT OUTCOME

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

39 The synthesis of haem

Mitochondria Ferrochelatase dback Haem e fee tiv ga Ne Fe 2+

Glycine Protoporphyrin IX + (PpIX) Succinyl-CoA δ-aminolevulinic acid

synthase O OH H2N O δ-aminolevulinic acid Cytosol (ALA) Protoporphyrinogen III

Porphobilinogen Coproporphyrinogen III

Porphobilinogen deaminase UNIVE D RS N IT U Y L MEDICAL Uroporphyrinogen III

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Benefits & Disadvantages of PDT

Superficial PDT + Selective treatment + Good cosmetic results + Fast healing + Repeated many times + Outpatient basis + “Simple” method (cooling with water as anaesthetics, occasionally) – Thin and superficial tumours

…compared to conventional methods

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

40 Benefits & Disadvantages of I-PDT

Interstitial PDT

+ Large and/or deeply lying tumours

– Mechanical damage – Insertion of fibres – Dosimetry

...compared to superficial PDT UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Concept Action: InsertionStartContinueMeasureTreatment treatment fluence oftreatment finished optical rate fibres in predetermined Recalculatepositions treatment time

Tumour Healthy tissue

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

41 Calculations of delivered light dose

Not fully treated tumour tissue

Treated tumour tissue

High dose surrounding tissue

UNIVE D RS 20092 minutes N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Benefits & Disadvantages of IPDT

Interstitial PDT

+ Large and/or deeply lying tumours

– Minimally invasive – Positioning of the fibres – Dosimetry

...compared to superficial PDT UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

42 Our patented technology

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Dosimetry measurements during treatment

Fluence rate - fibre positions - optical feedback Fluorescence - build-up of PpIX - PpIX bleaching Temperature -IR camera Tissue perfusion - laser Doppler imaging

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

43 PpIX photobleaching

Pre PDT 40361218251 minutesminute Fluorescence intensity (a.u.)

Excitation wavelength

UNIVE D RS N IT U Y 400 500 600 700 L MEDICAL Wavelength (nm)

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Temperature measurements

• Monitor the heating during PDT

• Observe vascular effects

• Thermal imaging camera (AGEMA

• Resolution 0.1 ºC

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

44 Temperature measurements

Basal cell carcinoma on the chest. 127 mW/cm2, 60 J/cm2

PDT fiber

Pre PDT 3 cm 140 s 33,6ºC 36,4ºC

Temp. measurement area

320 s 480 s, post PDT 37,0ºC 36,7ºC

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Temperature measurement

Temperature increase (ºC) Laser off Basal cells cancer 4 127 mW/cm2

3 Chest, ALA Back, ALA-ME 2

1

0 0 100 200 300 400 500 600 UNIVE D RS N IT U Y L MEDICAL Time (seconds)

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

45 Laser Doppler Perfusion Imaging

Laser

λem: 633 nm

Scanning

Detector Signal

UNIVE D RS Processing N IT U Y L MEDICAL Computer

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Laser Doppler Perfusion Imaging

Pre PDT Im. post PDT 1 w post PDT

1.52 0.00 2.30 0.00 0.51 0.10 Tissue perfusion (a. u.) - PDT 1 m post PDT1.2 13 m post PDT

Superficial BCC 1.0 Normal tissue

0.8

0.6

0.4

UNIVE D RS N IT U Y L 0.2 MEDICAL 0.0 LASER CENTRE Lund University Medical Laser Centre, SwedenPre Im post 1w 1m 2m 3m 13m 1991 0.91 0.08 1.73 0.37

SWEDEN

46 Set-up for spatially resolved diffuse reflectance

Lamp Mirror log(I/I0) Fibre CCD -25 0 25 r(mm)

Tissue

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Diffuse Reflectance from Skin at 630 nm

log(I/I0)

-25 0 25 r(mm)

Diffuse reflectance of normal skin irradiated with a pencil beam UNIVE D RS N IT U Y L MEDICAL of light at 630 nm, measured as line across the irradiated spot.

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

47 Models and methods

- Spatially resolved CW reflectance

Constant µ’s Constant µa

Optimal distances : one at the “Hinge” and one beyond the “Pivot”

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Spatially resolved CW measurements

Diffusion equation - curve fitting Monte Carlo method - curve fitting Calibration against known standards

r

tissue

μa , μs’

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

48 Results Fit of MCS to diffuse reflectance

120 profile

100

80

60

40 intensity (CCD count) (CCD intensity

Intensity (CCD counts) (CCD Intensity 20

0 0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0 ρρ (mm)

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

Fit versus expected optical Results properties 0.06 1.20

0.05 1.05

) 0.04 ) -1 ) ) -1 -1 -1 0.03 0.90 (mm ' (mm (mm s a a μ μ ’ (mm μ s

0.02 μ 0.75 0.01

0.00 0.60 550 600 650 700 750 800 850 550 600 650 700 750 800 850 Wavelengthwavelength (nm) (nm) Wavelengthwavelength (nm) (nm)

UNIVE D RS N IT U Y L MEDICAL

LASER CENTRE Lund University Medical Laser Centre, Sweden 1991

SWEDEN

49