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 1

•  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.


RAT BRAIN : Different zoom levels

16x13mm2

4x4mm2

1x1mm2

Advanced Imaging in Bio


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


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


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


27 © LLTECH 2011

Zoom on basal cell carcinoma: discriminaIon at cellular level

200 µm

100 µm

Peritumoral stroma

High cell density


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

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

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

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

57 © LLTECH 2011 57

Leaf veins – fresh leaf sample

© LLTech 2012

Fibers

Veins

100 µm

500 µm

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



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…


63 © LLTECH 2011

LLTech

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www.lltechimaging.com [email protected]


lltech light-ct scanner image atlas

Health & Medicine