Atlas of Light-CT Images

LLTech Light-CT Scanner Atlas of images 2012
LLTech 2012 www.lltechimaging.com - No copy without authorization
TABLE OF CONTENTS Table of contents

BREAST BRAIN SKIN LUNG GI URO GENITAL NEEDLE CORE THYROID Page 6 Page 24 Page 34 Page 58 Page 64 Page 69 Page 78 Page 83 FAT CELLS CORNEA AND RETINA PLANT IMAGING CONTACTS Page 86 Page 88
DEVELOPMENT BIOLOGY Page 99 Page 112 Page 118
Light-CT key benefits
Optical in-depth biopsies of gross tissue within minutes
1 m 2D and 3D histopathological resolution

Easy exploration, acquisition and rendering in DICOM format Safe, non-invasive and non-destructive process
Fast and non-invasive 3D in-depth structural and cellular imaging
Light Computed Tomography technology
Based on Full-Field Optical Coherence Tomography (FFOCT)

Combines microscope resolution with interferometry High resolution in-depth C scans
Commercial device specs:

Excellent resolution: 1.5m transverse, 1m axial 70Hz max. tomographic frame rate 0.8 mm x 0.8 mm
Penetration depth 200m 1mm depending on tissue scattering

25 mm diameter sample size Small footprint: Scanner and light source fit on 70 cm x 35 cm
Light-CT Scanner for ex-vivo cellular imaging
User friendly acquisition software DICOM 2D and 3D Viewer
Optical acquisition unit, moving vertically
Movable tray with sample holder X,Y moving stage
White Light Source
Joystick for easy control of X,Y, Z movements
Integrated wide field camera to take sample picture before imaging

5
BREAST IMAGING
BREAST IMAGING
Back to table of contents
Healthy breast tissue
Grainy aspect of normal fibrous tissue
Duct with calcification
Lobule
Adipocytes
Vessel
Courtesy of Hpital Tenon, Paris, France 7
Breast tissue diagnosis decision tree
LLTech 2012
Figure 3 : DiagnosisLLTech 2012 www.lltechimaging.comon human breast tissue decision tree for FF-OCT images - No copy without authorization
Breast Histology DCIS Ductal Carcinoma in situ

Fat cells
1 cm
Lobules
Breast., Ductal Carinoma H&E vs LightCT on Fresh Tissue Hopital Tenon, France, August 2010
LLTech 2012 www.lltechimaging.com - No copy without authorization 9
Breast Histology DCIS Ductal Carcinoma in situ

Lobules
Duct with necrosiss
10
Breast Histology Invasive adenocarcinoma - Nodular tumor
Healthy tissue: grainy medium scattering fibers
Tumorous tissue: highly scattering thin trabeculae aspect of fibers

Breast Milk duct with thick wall 1 mm
3 mm
1 2
Connective tissue Fat cells
1 mm
Blood vessels
Milk Duct LLTech 2012 www.lltechimaging.com - No copy without authorization
Traditional Histology
12
Breast Histology Identification of the milk duct structures
Milk Duct End
Fat Cells
13
Healthy breast tissue
Lobule
Galactophorous duct cut longitudinally
Honeycomb configuration of adipocytes

LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of Hpital Tenon, Paris, France 14
Nipple - Cross section
Collecting duct with secretion in the lumen
Collecting duct in the retroareolar section

Invasive adenocarcinoma with in Situ Component
Highly scattering thin trabeculae aspect of fibrous tissue surrounding grey cellular zones
LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization
Enlarged ducts filled with cells proliferation or necrosis
Fibroadenoma
Well delimited nodule with lobulated appearence
Elongated compressed ductules with slit-like lumen

Fibroadenoma
Enlarged ductules characteristic of the lesion
LLTech 2012
Normal Sentinel Node
Adipocytes in the center of the node
Vessel Lymphoid zone of the node
LLTech 2012
Normal Sentinel Node
Germinal centers: lumphoid tissue
Trabeculae
Subcapsular sinus
Capsule
Invaded Sentinel Node
Fibrous enveloppe of the node Normal lymphoid zone with follicles (Dark grey)
Metastatsis (Light grey)
Hypervascularization due to metastasis

The Light-CT scanner allows fast tissue processing and pathology examination. It completes the toolset available to pathologists Tissue Scanner
Biopsy
5-8 minutes Non-Destructive 2D Digital Image
Slide
3D Digital Image
15-12 mins Artefacts Destructive
When Frozen Section is needed (Mostly Intra-operative)
12-24 hours Expensive Destructive
When Chemical Fixation is needed (Mostly Diagnostic)
22
Light-CT could help to reduce the number of re-excisions in breast cancer surgery whilst preserving the current routines of pathologists
Per surgery
Post surgery
Up to 40% of double surgical procedures
Current techniques Up to 40% of double procedures
cancerous
Cancerous
Surgery
Visual analysis, cryostat

Node
OK
Histology And/or Immuno histochemistry
OK
Further treatment
Poor identification of metastasis
No surgical margins analysis

Margin
Few minutes analysis Surgeon Pathologist
Several day analysis Pathologist
Per surgery
Cancerous
Post Surgery
Decrease of double surgical procedures
Cancerous
LLTech Light-CT Significant decrease of double procedures
Surgery
LLTechs analysis
OK
Histology And/or Immuno histochemistry
OK
Further treatment
Surgical margin analysis Metastasis analysis (> 2mm) et micro metastaisi : PN1mi > 0.2 2 mm <
Few minutes analysis Surgeon
Several day analysis

23
Pathologist Pathologist LLTech 2012 www.lltechimaging.com - No copy without authorization
BRAIN IMAGING
24
Rat brain
500 m
1 mm
Architectural and cellular information seen at various scales Myelinated fibers visible
100 m
Human epileptic parenchyma
Cortex
Axons bundles
50m
Neurons cell bodies (dark dots)
50m
White matter
Axons/myelin fibers
500m
50m
Capillary
Hippocampus : Coronal section (left side)
Pyramidal neurons of the CA4 field

50m
CA4 Field
Hippocampal sulcus
100m
500m
Stratum radiatus of the CA1 field

Stratum granulosum of the gyrus dentatus
CA1 field of the cornus ammonis
27
Fibroblastic meningioma grade I
500m
200m
100m
200m
Wide fascicles of tumour cells

LLTECH 2011
Large capillary
Collagen-rich matrix
28
Transitional meningioma grade I
100m
Collagen-rich matrix
50m
Whorls formation
50m 500m
Psammoma surrounding a calcification
29
Choroid Plexus Papilloma : Xanthomatous changes
Characteristic cauliflower-like masses
Papilla filled with blood
Empty papilla
Spinal cord : normal tissue
100 m
Arrows: Fibers transverse cut
500 m
100 m
LLTECH 2011
31
Spinal cord with Iysolecithin-induced injury
Lesion
500 m
32
Injured spinal cord longitudinal cut
Injured area: - disorganized architecture - no visible regular fiber arrangement - modified contrast
100 m
Healthy area: - regular architecture - longitudinally packed fibers
100 m
33
SKIN IMAGING
34
Normal skin morphology
a
Epidermis
Collagen
500 m
Pilosebaceous unit
Sweat gland
Adipocytes
LLTech 2012
Courtesy LLTech 2012 www.lltechimaging.com - No copy without authorization of Hopitaux Universitaires de Genve, Genve, Switzerland 35
Skin morphology: layers are well discriminated

Epidermis
3
1
Dermis
2 4
Hypodermis
5 mm x 8 mm: mosaic of native field images Ageing patient (solar elastosis) En-face imaging of vertical excision
Structures are well discriminated with structural and cellular details

3 Pilosebaceous unit 1 - Epidermis
Stratum corneum
KN KN SG
HF: hair follicle SG: sebaceous gland

HF SG
KN
Stratum spinosum
Stratum basale 100 m KN: keratinocyte nuclei
SG
SG
200 m
2 - Dermis 4 - Sweat gland

Collagen
Adipocytes
200 m
100 m
37
En-face skin slicing shows structural and cellular details
Corneocytes
Keratinocyte nuclei
Epidermis
100 m
100 m
Stratum corneum
Stratum spinosum
Melanin papillary caps
Blood vessels
Dermis
Collagen
100 m
100 m
Stratum basale
LLTech 2012
Reticular region
Skin vertical reconstruction from en-face slicing for layers assesment and measurement
KN: Keratinocyte nuclei BV: Blood vessels

KN
Papillary melanin caps
Stratum corneum
KN KN
Stratum spinosum
BV BV BV
50 m
Dermis
LLTech 2012
3D reconstruction from en-face slicing
Epidermis and dermis separated by sodium bromide

Epidermis 800 m x 800 m Dermis 800 m x 800 m
Wrinkle
Stratum corneum
Stratum spinosum
Dermal papilla
Collagen
Blood vessel
50 m
50 m
Reconstructed vertical slice
Reconstructed vertical slice
LLTech 2012
Vertical reconstruction of skin model
Keratinocyte nuclei Stratum corneum Stratum spinosum
Dermis
50 m
LLTech 2012
41
Stratum Corneum
Reconstructed z slices
50 um
42
Normal skin
a
500 m
f e
100 m
Normal skin zoom
Stratum corneum
Stratum spinosum
Dermis
Collagen - dermis
Adipocytes
Deep dermis
44
Adipocytes
200 m
9um depth
27um depth
60um depth
Reconstructed depth slice

50 m
Skin pathologies: basal cell carcinoma discrimination at structural level
Dense peritumoral stroma
LLTech 2012
Skin pathologies: basal cell carcinoma discrimination at structural level

Epidermis
Dermis
PSU
BCC
A C
Hypodermis
A 1 mm
PSU
C
BCC: Basal cell carcinoma A: Adipocytes C: Collagen PSU: Pilosebaceous unit

LLTech 2012 Courtesy LLTech 2012 www.lltechimaging.com - No copy without authorization of Hopitaux Universitaires de Genve, Genve, Switzerland 47
Zoom on basal cell carcinoma: discrimination at cellular level
Peritumoral stroma
High cell density
200 m
100 m
LLTech 2012
Skin pathologies: discrimination of actinic keratosis
Enlarged epidermis
1 mm
Extended epidermal ridges

LLTech 2012 Courtesy LLTech 2012 www.lltechimaging.com - No copy without authorization of Hopitaux Universitaires de Genve, Genve, Switzerland 49
Basal cell carcinoma: in depth imaging

SG: Sebaceous glands
BCC SG
Z = 10 microns, 7 x 4.2 mm FOV
Application to Mohs surgery
LLTech 2012
Healthy vs. Cheloid scar tissue
Healthy skin tissue FOV: 7.5x5mm
Cheloid scar tissue: FOV: 6.5x6.5mm
LLTECH 2011 LLTECH 2011
51
Cheloid scar tissue in detail
Dermo-epidermal junction
500 m
1mm
Thick collagen fiber bundles
Nodular structure
600 m
LLTECH 2011
Cheloid scar tissue viewed en face
20m depth
120m depth
53
Skin in-vivo: fingerprint imaging in 3D
Fingerprints 3D reconstruction FOV 800 m x 800 m Sweat ducts
Corneocytes
100 m
En-face image of fingerprints
54
Skin in-vivo: sweat duct imaging in 3D
SD: sweat ducts
SD
SD
Stratum corneum
100 m
Stratum spinosum
Vertical reconstruction from en-face images
Sweat duct 3D reconstruction

Skin in-vivo: skin layers imaging en face and in vertical reconstruction
Stratum corneum
Stratum spinosum
Dermis
Stratum corneum Stratum spinosum
Dermis
100 m
Evaluation of stratum corneum thickness: 15 m
56
Mouse skin in vivo

10um depth 20um depth
En-face slices in thigh region

FOV 1mm2 Penetration to 200um
Hairs near surface
35um depth
85um depth
Hair follicles Collagen fibers
LLTech 2012
Courtesy of Institut Langevin, Paris,57 France
LUNG IMAGING
58
Human Lung
Limit of cancerous stroma / alveoli with macrophages
Zone 2
Alveoles
Zone 4
Zone 1
Zone 3
Zone with cancerous tissue
500m
LLTech 2012
Courtesy of Hpital Hotel-Dieu, Paris, France 59
Human Lung Zone 1 Zoom on cancerous stroma
Stroma
Cancerous area
200m
LLTech 2012
Human Lung Zone 2 Limit cancerous stroma / alveoli with visible macrophages and vessel
Alveoli
Macrophages
Vessel
LLTech 2012
Human Lung Zone 3 Limit cancerous stroma / alveoli with visible macrophages
Macrophages
100m
LLTech 2012
Human Lung Zone 4 Alveoli
Alveoli
200m
LLTech 2012
GI IMAGING
64
Human colon normal tissue
X 20
1mm
X 100
100 m
LLTECH Few minutes

LLTech 2012 LLTECH 2008
Traditional histology Several days

Courtesy of institut Curie, Paris, France 65
Colonic carcinoma Colonic Carcinoma
LLTech 2012
Courtesy of MD Anderson, Houston, USA 66
Intestines Crohns disease
Crohns disease structure not visible from the surface
Fat cells
200 m
LLTech 2012
Courtesy of Institut Curie, Paris, France 67
Rat stomach
Mucosa with gastric pits
Submucosa with bright collagen fibers
Muscularis propria with muscle bundles
200 m
LLTech 2012
Courtesy of Weill Cornell Medical College, NY, USA 68
UROGENITAL IMAGING
LLTECH 2011
69
Rat bladder
Lamina propria
100 m
Stratified urothelium
Muscular bundles of the lamina propria
Serosa
100 m 500 m
LLTech 2012
Human bladder: low grade papillary urothelial carcinoma
100 m
Lesion with fibrovascular cores (arrow) and superficial umbrella cells (arrowheads)
LLTech 2012
Human bladder
1 mm
LLTech 2012
Courtesy of Hpital Tenon, without authorization LLTech 2012 www.lltechimaging.com - No copy Paris, France
100 um
72
Rat prostate
Prostatic acini
200 m
1 mm
Papillary folds of the acinar glands
Periprostatic fat
Fibromuscular stroma
Blood vessel
LLTech 2012
Mouse kidney
Renal cortex
Renal medulla
Renal sinus
1 mm
LLTech 2012
Courtesy of Institut Biomdical de Bictre, Paris, France 74
Mouse kidney zoom on highlighted section of previous slide
800m
LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of Hpital Kremlin Bictre, Paris,75 France
Human kidney
800m
200m
200m
LLTech 2012
Courtesy of Hpital Tenon, Paris,76 France
Human kidney
800m
Courtesy of Hpital Tenon, Paris, France
400m
300m
LLTech 2012
Courtesy of Hpital Tenon, Paris,77 France
CORE NEEDLE BIOPSIES
LLTECH 2011
78
Kidney
100m
Renal tubules
500m
50m
Vessel
LLTech 2012
Breast : Invasive lobular carcinoma

adipocytes
stroma
500m
LLTech 2012
Breast: Infiltrative carcinoma
LLTech 2012
Breast: Ductal carcinoma in situ
500m
LLTech 2012
THYROID IMAGING
LLTECH 2011
83
Thyroid
200 m
800 m
LLTech 2012
Thyroid
200 m
800 m
LLTech 2012
FAT CELLS IMAGING
LLTECH 2011
86
Brown fat cells - adipocytes

En face slices
Adipocytes
100 m
100 m
200 m
LLTech 2012
Courtesy of Centre de Recherche des Cordeliers, Paris, France LLTech 2012 www.lltechimaging.com - No copy without authorization 87
CORNEA AND RETINA
LLTECH 2011
88
Human cornea reconstructed depth slice
50 um
www.lltechimaging.com
89
Large field en face view of epithelial layer
100 um
200 um
90
Large field en face view in stroma
200 um
91
Oedematous cornea
50 um
92
Rabbit cornea after LASIK: reflectivity profiles reveal increased scattering and thickened epithelial layer
LASIK
LASIK
CONTROL
CONTROL
LLTech 2012
93
Newborn mouse anterior segment
Corneal epithelial cells Basal membrane Stroma
Descemets membrane
Aqueous humor
Lens capsule Lens epithelial cells Cortical fibers
LLTech 2012 LLTech 2012
Courtesy of ESPCI, Paris,94 France
Rat retina: comparison with histology
RPE PRL
ONL
OPL INL
IPL
Figure 1: Right: Ex vivo rat retina imaged using the full field OCT system of ESPCI. Left: Histology for comparison.
GCL NFL
Field: 230x300m
Invest Ophthalmol Vis Sci. 2004 Nov;45(11):4126-31.
Grieve K, Paques M, Dubois A, Sahel J, Boccara C, Le Gargasson JF.

LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization 95
Rat retina
Nerve fiber layer
Transscleral image
Retina
Invest Ophthalmol Vis Sci. 2004 Nov;45(11):4126-31.
Grieve K, Paques M, Dubois A, Sahel J, Boccara C, Le Gargasson JF.

LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of ESPCI, Paris,96 France
Pig retina
Photoreceptors - pig
Retinal pigment epithelium - pig
Arterio-venous crossing Pig
LLTech 2012
97
Optic Nerve - rat
LLTech 2012
98
DEVELOPMENTAL BIOLOGY
LLTECH 2011
99
In vivo imaging of drosophila melanogaster
LLTech 2012
Courtesy of ESPCI, Paris, France 100
In vivo imaging of drosophila melanogaster four stages of pupation over 72 hours
Prepupa (0-2 h)
Transition to pupal stage (24 h)
Pupal stage (48 h)
Advanced pupal phase before eclosion (72 h)
LLTech 2012
Pupa 118: 4 days 30um depth

100 um
102
Drosophila
200 m
LLTech 2012
C. Elegans Fast identification of anatomy
15 m depth
30 m depth
ovocytes intestine Body wall - cuticle gonad arm
uterus
100 m
100 m
LLTech 2012
Courtesy of ENS, Paris, France 104
C. elegans Images at higher magnification: 40X
100 m
3D reconstruction
LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization
50 m
Courtesy of ENS, Paris, France 105
Zebra fish eye

En face slices
100 m
10um depth
20um depth
45um depth
100 m

LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of Inserm, Paris, France 106
Xenopus Laevis
Cartilage Heart
100 um
107
Xenopus Laevis: back/tail

100 um
108
Xenopus Laevis eye: en face slices
Surface
10um depth
20um depth
30um depth
40um depth
50um depth
60um depth
70um depth
80um depth
90um depth
100um depth
110um depth
200 um
109
Mouse embryo in vitro 6.5 days old
3D imaging of internal cavity
LLTech 2012
France LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of Ellvire Guiot and nstitut Jacques Monod, Paris,110
Sabellaria alveolata: mouth palps
100 m
Mouth palps
500 m
LLTech 2012
Courtesy of Muse National dHistoire Naturelle, Paris, France LLTech 2012 www.lltechimaging.com - No copy without authorization 111
PLANT IMAGING
LLTECH 2011
112
Mouse ear leaf epidermis
Epidermal cell
Stoma
100 m
LLTech 2012
113
Leaf veins
Veins
500 m
Fibers
100 m
114
Apple
Wax Skin
Water
Flesh
500 m
100 m
115
Effect of harpin infiltration on tobacco leaf

10 min after harpin infiltration 30 min after harpin infiltration
Upper epidermis
Epidermis cell
50 m
50 m
Palisade parenchyma
Chloroplast
50 m
50 m
116
LLTech 2012 LLTech 2012 www.lltechimaging.com - No copy without authorization Courtesy of UPMC, Paris, France 116
Light-CT does not require tissue preparation, nor staining of any kind Creates images within minutes using a non-destructive process Offers a 1 m cellular resolution in 3D Reveals structural and cellular information Mosaicing allows fast visualisation at various scale En-face high-resolution imaging allows vertical and 3D reconstruction In-vivo images show good promise for in-vivo system Clinical and research applications in cancer detection, cosmetics, biobanking, small animal, developmental biology, botany
117
Contact Details
USA
Europe
LLTech Inc. 103 Carnegie Center Drive Suite 300 Princeton, NJ 08540 USA www.lltechimaging.com contact@lltech.fr Phone :+ 1 609 955 3506
LLTech SAS 29 rue du Faubourg St-Jacques Ppinire Paris Sant Cochin 75014 Paris - France www.lltechimaging.com contact@lltech.fr Phone :+33 1 82 72 61 25
118

Atlas of Light-CT Images

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Atlas of Light-CT Images

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LLTech Light-CT Scanner Atlas of images 2012

LLTech 2012 www.lltechimaging.com - No copy without authorization

TABLE OF CONTENTS Table of contents

DEVELOPMENT BIOLOGY Page 99 Page 112 Page 118

LLTech 2012 www.lltechimaging.com - No copy without authorization

Light-CT key benefits

Optical in-depth biopsies of gross tissue within minutes

1 m 2D and 3D histopathological resolution

Fast and non-invasive 3D in-depth structural and cellular imaging

LLTech 2012 www.lltechimaging.com - No copy without authorization

Light Computed Tomography technology

Based on Full-Field Optical Coherence Tomography (FFOCT)

Commercial device specs:

Penetration depth 200m 1mm depending on tissue scattering

LLTech 2012 www.lltechimaging.com - No copy without authorization

Light-CT Scanner for ex-vivo cellular imaging

User friendly acquisition software DICOM 2D and 3D Viewer

Optical acquisition unit, moving vertically

Movable tray with sample holder X,Y moving stage

White Light Source

Joystick for easy control of X,Y, Z movements

LLTech 2012 www.lltechimaging.com - No copy without authorization

Integrated wide field camera to take sample picture before imaging

Back to table of contents

LLTech 2012 www.lltechimaging.com - No copy without authorization

Healthy breast tissue

Grainy aspect of normal fibrous tissue

Duct with calcification

LLTech 2012 www.lltechimaging.com - No copy without authorization

Breast tissue diagnosis decision tree

Breast Histology DCIS Ductal Carcinoma in situ

Breast Histology DCIS Ductal Carcinoma in situ

Duct with necrosiss

LLTech 2012 www.lltechimaging.com - No copy without authorization

Breast Histology Invasive adenocarcinoma - Nodular tumor

Healthy tissue: grainy medium scattering fibers

Tumorous tissue: highly scattering thin trabeculae aspect of fibers

Breast Milk duct with thick wall 1 mm

Breast Histology Identification of the milk duct structures

Milk Duct End

LLTech 2012 www.lltechimaging.com - No copy without authorization

Healthy breast tissue

Galactophorous duct cut longitudinally

Honeycomb configuration of adipocytes

Nipple - Cross section

Collecting duct with secretion in the lumen

Collecting duct in the retroareolar section

Invasive adenocarcinoma with in Situ Component

Enlarged ducts filled with cells proliferation or necrosis

Courtesy of Hpital Tenon, Paris, France 16

Well delimited nodule with lobulated appearence

Elongated compressed ductules with slit-like lumen

Enlarged ductules characteristic of the lesion

LLTech 2012 www.lltechimaging.com - No copy without authorization

Courtesy of Hpital Tenon, Paris, France 18

Normal Sentinel Node

Adipocytes in the center of the node

Vessel Lymphoid zone of the node

LLTech 2012 www.lltechimaging.com - No copy without authorization

Courtesy of Hpital Tenon, Paris, France 19

Normal Sentinel Node

Germinal centers: lumphoid tissue