LLTECH LIGHT-CT SCANNER IMAGE ATLAS
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Transcript of LLTECH LIGHT-CT SCANNER IMAGE ATLAS
1 © LLTECH 2011
Atlas of images February 2013
www.lltechimaging.com
2 © LLTECH 2011
• BREAST Page 10
• BRAIN Page 15
• SKIN Page 21
• LUNG Page 31
• KIDNEY Page 35
• EYE Page 39
• NEEDLE CORE Page 45
• EMBRYOLOGY Page 48
• PLANTS Page 56
TABLE OF CONTENTS
3 © LLTECH 2011
The Light-‐CT scanner allows fast >ssue processing and pathology examina>on. It completes the toolset available to pathologists
When Chemical FixaIon is needed (Mostly DiagnosIc)
When Frozen SecIon is needed (Mostly Intra-‐operaIve)
Biopsy 3D
Digital Image
2D Digital Image
Slide
15-‐12 mins Artefacts DestrucIve
12-‐24 hours Expensive DestrucIve
5-‐8 minutes Non-‐DestrucIve
Tissue Scanner
4 © LLTECH 2011 © LLTECH 2011 © LLTECH 2012 – www.lltechimaging.com
Imaging in sca]ering media: techniques
u LightCT
5 © LLTECH 2011 © LLTECH 2011 © LLTECH 2012 – www.lltechimaging.com
Light-‐CT scanner
OpIcal acquisiIon unit, moving verIcally • User friendly
acquisiIon soaware
• DICOM 2D and 3D Viewer
Movable tray with sample holder
X,Y moving stage
JoysIck for easy control of X,Y, Z movements
White Light Source
Integrated wide field camera to take sample picture before
imaging
• OpIcal in-‐depth biopsies of gross Issue within minutes
• 1 µm 2D and 3D histopathological resoluIon
• Easy exploraIon, acquisiIon and rendering in DICOM format
• Safe, non-‐invasive and non-‐destrucIve process
Light-‐CT™ key benefits
Fast and non-‐invasive 3D in-‐depth structural and cellular imaging
Based on Full-‐Field OpIcal Coherence Tomography (FFOCT)
Combines microscope resoluIon with interferometry
High resoluIon in-‐depth C scans
Commercial device specs:
• Excellent resoluIon: 1.5µm transverse, 1µm axial
• 70Hz max. tomographic frame rate – 0.8 mm x 0.8 mm
• PenetraIon depth 200µm – 1mm depending on Issue sca]ering
• 25 mm diameter sample size
• Small footprint: Scanner and light source fit on 70 cm x 35 cm
Light CT technology
8 © LLTECH 2011
Image acquisiIon
§ Put sample into holder
§ Put sample holder into
tray
• The system takes a wide field picture
1.Prepare sample
2. Select area to be scanned
3. AutomaIc scanning
4. Analyze result in the viewer
• The user selects the area to be scanned from the wide field image
• The user launches the automaIc scanning
• Results are analyzed in the DICOM viewer
• 2D and 3D rendering • DICOM image database • Export to various formats: DICOM, TIF, JPEG, GIF…
• MulIple languages for image viewer (English, Arabic, French…) Fast and easy process: 1 cm2 in 5-‐7 minutes
9 © LLTECH 2011
PATHOLOGY APPLICATIONS INTRAOPERATIVE DIAGNOSIS
10 © LLTECH 2011
BREAST AND LYMPH NODES
11 © LLTECH 2011
Grainy aspect of normal fibrous Issue
Duct with calcificaIon
Lobule Adipocytes Vessel
Healthy breast Issue
© LLTech 2012 Courtesy of Hôpital Tenon, Paris, France
Assayag et al, TCRT Express 2013 in press
12 © LLTECH 2011
100µm
Highly scaHering thin trabeculae aspect: fibrous >ssue in the tumourous part
Grainy medium scaHering aspect: healthy fibrous >ssue
INVASIVE ADENOCARCINOMA : NODULAR TUMOR
© LLTECH 2011
Interface of the tumourous and normal appearing part :
Different aspect of the fibrous >ssue
Grey zones corresponding to foci of carcinomatous cells
Tumour margin
500µm
500µm SENSITIVITY= 97% SPECIFICITY=74%
13 © LLTECH 2011 © LLTECH 2011
Nodules containing lymphoid >ssue
Normal Sen>nel Node
Adipocytes in hilum
Capsule
14 © LLTECH 2011
Lymphoid zone
Invaded zone
Invaded Sen>nel Node
Bright white aspect, highly scaHering dense collagen meshwork
SENSITIVITY= 100% SPECIFICITY=97%
Grieve et al Proc SPIE 2013
15 © LLTECH 2011
BRAIN AND SPINAL CORD
16 © LLTECH 2011
Neuron cell bodies (dark dots)
Axons/myelin fibers
Capillary
Cortex
White maHer
Axon bundles
FF-‐OCT imaging of human epilep>c brain and cerebellum iden>fies myelinated axon fibers, a subpopula>on of neuronal cell bodies and CNS vasculature.
Harms et al Proc SPIE 2013
17 © LLTECH 2011
CA4 field
Hippocampal sulcus
Alveus
Sub-‐ Ependymal zone
CA1 field of the cornus ammonis
Stratum radiatus of the CA1 field
Stratum granulosum of the dentate gyrus
Pyramidal neurons of the CA4 field responsible for memory
Hippocampus: Coronal sec>on (led side)
18 © LLTECH 2011
FF-‐OCT images dis>nguish meningiomas from meningeal haemangiopericytoma
Collagen bundles
Whorls
CalcificaIon
Collagen balls
Meningioma psammoma – diagnosis possible directly on FF-‐OCT image
80% of meningiomas are benign. Histologically, meningioma cells are relaIvely uniform, with a tendency to encircle one another, forming whorls and psammoma bodies (laminated calcific concreIons).They have a tendency to calcify and are highly vascularized. Areas of fibrosis may be present.
19 © LLTECH 2011 © LLTECH 2011
RAT BRAIN : Different zoom levels
16x13mm2
4x4mm2
1x1mm2
Advanced Imaging in Bio
20 © LLTECH 2011 © LLTECH 2011
Arrows: Fibers transverse cut
Spinal cord : normal Issue
500 µm
100 µm
100 µm
21 © LLTECH 2011
DERMATOLOGY AND COSMETICS
22 © LLTECH 2011
Normal skin morphology
Epidermis
Collagen
Pilosebaceous unit
Sweat gland
Adipocytes
b
a
c
e
d
500 µm
© LLTech 2012 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland
Dalimier and Salomon Dermatology 224, 84-92 (2012)
23 © LLTECH 2011
Skin verIcal reconstrucIon from en-‐face slicing for layers assesment and measurement
Stratum corneum
Stratum spinosum
Dermis
Papillary melanin caps BV: Blood vessels
BV
BV BV
KN KN KN
KN: KeraInocyte nuclei
50 µm
© LLTech 2012 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland
50 µm
Stratum corneum
Stratum spinosum
Dermis
KeraInocyte nuclei
VerIcal reconstrucIon of skin model
24 © LLTECH 2011
3D reconstrucIon from en-‐face slicing
Wrinkle Stratum corneum Stratum spinosum
Epidermis
800 µm x 800 µm
Dermis
Dermal papilla Collagen
800 µm x 800 µm
Epidermis and dermis separated by sodium bromide
Blood vessel
50 µm
Reconstructed verIcal slice
50 µm
Reconstructed verIcal slice
© LLTech 2012 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland
25 © LLTECH 2011
Adipocytes
9um depth 27um depth 60um depth
Reconstructed depth slice
200 µm
50 µm
26 © LLTECH 2011
Skin pathologies: basal cell carcinoma discriminaIon at structural level
Dense peritumoral stroma
© LLTech 2012 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland
27 © LLTECH 2011
Zoom on basal cell carcinoma: discriminaIon at cellular level
200 µm
100 µm
Peritumoral stroma
High cell density
© LLTech 2012 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland
28 © LLTECH 2011
Skin in-‐vivo: fingerprint imaging in 3D
100 µm
Sweat ducts
Fingerprints 3D reconstrucIon FOV 800 µm x 800 µm
Corneocytes
En-‐face image of fingerprints
LightCT arm support add-‐on allows in vivo imaging on arm, hand, fingers
29 © LLTECH 2011
Skin in-‐vivo: skin layers imaging en face and in verIcal reconstrucIon
Stratum corneum Stratum spinosum Dermis
Stratum corneum
Stratum spinosum
Dermis 100 µm EvaluaIon of stratum corneum thickness:
15 µm
30 © LLTECH 2011
Mouse skin in vivo En-‐face slices in thigh region FOV 1mm2
PenetraIon to 200um
© LLTech 2012 Courtesy of InsItut Langevin, Paris, France
10um depth 20um depth
35um depth 85um depth
Hairs near surface
Hair follicles Collagen fibers
31 © LLTECH 2011
LUNG
32 © LLTECH 2011
Cross secIon of rat lung
Jain et al. J Pathol Inform 2011, 2:28. Scale bar = 0.5 mm
33 © LLTECH 2011
Alveoli Blood vessel
Image from non-‐neoplasIc human lung Issue
FOV: FFOCT ~ 4 mm x 4 mm; H&E = 4X
Collapsed lung
Jain et al. Pathology Visions 2013.
34 © LLTECH 2011
Adenocarcinoma of lung with lepidic & invasive components
Lepidic
Invasive Invasive
Lepidic
Invasive
FOV: FFOCT ~ 4 mm x 4 mm; H&E = 4X . Insets: FFOCT ~ 0.375mm x 0.375mm; H&E = 20X
Invasive
35 © LLTECH 2011
KIDNEY
36 © LLTECH 2011
Blood vessel Glomerulus
Tubules 3xzoom (FOV ~0.19x0.19 mm)
3x zoom (FOV ~0.19x0.19 mm)
3x zoom (FOV ~0.19x0.19 mm)
Image from non-‐neoplasIc kidney Issue
FFOCT: FOV~ 0.37x0.25 mm, depth of imaging 0.1um
Glomerulus
Tubules
Blood vessel
Caron et al. Proc SPIE 2013
37 © LLTECH 2011
Blood vessel Glomerulus
Tubules
Glomerulus 20X
Tubules 20X
Blood vessel 20X
H&E = 10X
H&E image from non-‐neoplasIc kidney
Caron et al. Proc SPIE 2013
38 © LLTECH 2011 Chromophobe RCC
Sheets of cells
NeoplasIc kidney: normal architecture replaced by sheets of cells
Sheets of cells
Sheets of cells
3x zoom (FOV~ 0.19x0.19mm)
FFOCT: FOV ~ 0.38x0.38 mm, depth of imaging : 7.2um
H&E 10X
20X
39 © LLTECH 2011
RETINA AND CORNEA
40 © LLTECH 2011
ReIna
© LLTech 2012 Courtesy of ESPCI, Paris, France
Nerve fiber layer -‐ rat
ReIna -‐ rat
Grieve et al IOVS 2004 Nov;45(11):4126-31.
OpIc Nerve -‐ pig
Photoreceptors - pig
Retinal pigment epithelium - pig
41 © LLTECH 2011
Human graa cornea: over 500um thickness
50 um
www.lltechimaging.com Courtesy of Vincent Borderie Ghouali et al, ARVO 2013
42 © LLTECH 2011
Large field en face view of epithelial layer
200 um
100 um
www.lltechimaging.com Courtesy of Vincent Borderie
LightCT can evaluate integrity of epithelium and stroma in graa corneas (not possible with exisIng techniques)
43 © LLTECH 2011
200 um
Large field en face view in stroma
www.lltechimaging.com Courtesy of Vincent Borderie
ApplcaIon of LightCT in eye banks
44 © LLTECH 2011
LightCT shows corneal pathology and can penetrate even edematous cornea
Cornea with chemical burn – highly sca]ering upper stroma
Cornea with edema
45 © LLTECH 2011
CORE NEEDLE BIOPSIES
46 © LLTECH 2011
100µm
50µm
Core Needle Biopsy: Kidney
Renal tubules
Vessel
500µm
© LLTech 2012 Courtesy of InsItut Curie, Paris, France
LightCT provides an evaluaIon of Issue architecture in minutes
47 © LLTECH 2011
Core Needle Biopsy Breast: InfiltraIve carcinoma
© LLTech 2012 Courtesy of InsItut Curie, Paris, France
LightCT used intraoperaIvely on biopsy Issue provides fast, non destrucIve evaluaIon
48 © LLTECH 2011
DEVELOPMENTAL BIOLOGY
49 © LLTECH 2011
Developmental biology: Drosophila
200 µm
© LLTech 2012 Courtesy of ESPCI, Paris, France © LLTech 2012
50 © LLTECH 2011
In vivo imaging of drosophila melanogaster Four stages of pupaIon over 72 hours: automated Ime series
Prepupa (0-‐2 h) TransiIon to pupal stage (24 h)
Pupal stage (48 h) Advanced pupal phase before eclosion (72 h)
© LLTech 2012 Courtesy of ESPCI, Paris, France
51 © LLTECH 2011
Pupa 118: 4 days, 30um depth
100 um
LightCT can capture automated Ime series to follow in vivo development over a period of days
Burcheri et al, Proc SPIE 2013
52 © LLTECH 2011
OIc vesicle
CarIlage
Xenopus Laevis
100 um
Musée d’Histoire Naturelle
Pigmented cells
53 © LLTECH 2011
Xenopus Laevis Musée d’Histoire Naturelle
50 um
Reconstructed depth slices
54 © LLTECH 2011
Xenopus Laevis eye: en face slices
Surface 10um depth 20um depth 30um depth
40um depth 50um depth 60um depth 70um depth
80um depth 200 um
Musée d’Histoire Naturelle
230um depth 140um depth 180um depth
55 © LLTECH 2011
C. elegans – Images at higher magnificaIon: 40X
3D reconstrucIon 50 µm
100 µm
© LLTech 2012 Courtesy of ENS, Paris, France
56 © LLTECH 2011
PLANTS
57 © LLTECH 2011 57
Leaf veins – fresh leaf sample
© LLTech 2012
Fibers
Veins
100 µm
500 µm
Courtesy of ESPCI, Paris, France
58 © LLTECH 2011 58
Apple
© LLTech 2012
100 µm
Wax
Skin
Flesh
500 µm
Water
Courtesy of ESPCI, Paris, France
59 © LLTECH 2011
Sorghum: cross secIon
Wide field en face 14mmX15mm
Courtesy of Plate-forme d'Histocytologie et d'Imagerie
Cellulaire Végétale (PHIV) Montpellier RIO Imaging
60 © LLTECH 2011
Sorghum cross secIon: naIve field view
800um x 800um en face field
Reconstructed depth slices 500um sample thickness
Courtesy of Plate-forme d'Histocytologie et d'Imagerie
Cellulaire Végétale (PHIV) Montpellier RIO Imaging
61 © LLTECH 2011
Rice: hypocotyle
Wide field en face 5mmX2mm
Reconstructed depth slice, 200um pentraIon depth
Courtesy of Plate-forme d'Histocytologie et d'Imagerie
Cellulaire Végétale (PHIV) Montpellier RIO Imaging
62 © LLTECH 2011
Conclusion
• Full-‐field OCT does not require Issue preparaIon, nor staining of any kind
• Creates images within minutes using a non-‐destrucIve process
• Offers a 1 µm cellular resoluIon in 3D
• Reveals structural and cellular informaIon of normal and pathological Issue
• Mosaicing allows fast visualisaIon at various scales
• En-‐face high-‐resoluIon imaging allows verIcal and 3D reconstrucIon
• Possible applicaIons for biobanking, cancer detecIon, digital pathology…
www.lltechimaging.com
63 © LLTECH 2011
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