LLTECH LIGHT-CT SCANNER IMAGE ATLAS
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Document showing the imaging capabilities of the LLTech's Light-CT scanner. The Light-CT scanner has amazing imaging capabilities due to its ultra high resolution (1.5 X 1.5 X 1 µm). It offers the capability to do optical slices under the surface of the analyzed sample. The imaging process is fast, easy an safe for both the user and the sample. No staining is required.
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LLTECH LIGHT-CT SCANNER IMAGE ATLAS
- 1. Atlas of images February 2013 www.lltechimaging.com © LLTECH 2011 1
- 2. TABLE OF CONTENTS • 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 © LLTECH 2011 2
- 3. The Light-‐CT scanner allows fast >ssue processing and pathology examina>on. It completes the toolset available to pathologists Tissue Scanner 3D Biopsy Digital Image 5-‐8 minutes Non-‐DestrucIve 2D Digital Image Slide 15-‐12 mins When Frozen SecIon is needed Artefacts (Mostly Intra-‐operaIve) DestrucIve 12-‐24 hours When Chemical FixaIon is needed Expensive (Mostly DiagnosIc) DestrucIve © LLTECH 2011 3
- 4. Imaging in sca]ering media: techniques u LightCT © LLTECH 2011 © LLTECH 2012 – www.lltechimaging.com 4
- 5. Light-‐CT scanner OpIcal acquisiIon unit, • User friendly moving verIcally acquisiIon soaware • DICOM 2D Movable tray and 3D with sample Viewer holder X,Y moving stage White Light JoysIck for easy Integrated wide field camera to Source control of X,Y, Z take sample picture before movements imaging © LLTECH 2011 © LLTECH 2012 – www.lltechimaging.com 5
- 6. Light-‐CT™ key benefits • 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 Fast and non-‐invasive 3D in-‐depth structural and cellular imaging
- 7. Light CT technology 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
- 8. Image acquisiIon 1.Prepare 2. Select area to be 3. AutomaIc 4. Analyze result in sample scanned scanning the viewer § Put sample • The system • The user selects the • Results are analyzed in the into holder takes a wide area to be scanned DICOM viewer § Put sample field picture from the wide field • 2D and 3D rendering holder into image • DICOM image database tray • The user launches the • Export to various formats: automaIc scanning DICOM, TIF, JPEG, GIF… • MulIple languages for image viewer (English, Fast and easy process: 1 cm2 in 5-‐7 minutes Arabic, French…) © LLTECH 2011 8
- 9. PATHOLOGY APPLICATIONS INTRAOPERATIVE DIAGNOSIS © LLTECH 2011 9
- 10. BREAST AND LYMPH NODES © LLTECH 2011 10
- 11. Healthy breast Issue Assayag et al, TCRT Express 2013 in press Grainy aspect of normal Duct with Lobule Adipocytes Vessel fibrous Issue calcificaIon © LLTech 2012 © LLTECH 2011 Courtesy of Hôpital Tenon, Paris, France 11
- 12. INVASIVE ADENOCARCINOMA : NODULAR TUMOR Interface of the tumourous and normal appearing part : Different aspect of the fibrous >ssue Tumour margin 500µm Highly scaHering thin trabeculae aspect: fibrous >ssue in the tumourous part 100µm Grey zones corresponding to Grainy foci of carcinomatous cells medium scaHering aspect: healthy 500µm SENSITIVITY= 97% fibrous >ssue © LLTECH 2011 © LLTECH 2011 SPECIFICITY=74% 12
- 13. Normal Sen>nel Node Adipocytes in hilum Nodules containing lymphoid >ssue © LLTECH 2011 Capsule 13
- 14. SENSITIVITY= 100% Invaded Sen>nel Node SPECIFICITY=97% Invaded zone Lymphoid zone Bright white aspect, highly scaHering dense collagen meshwork Grieve et al Proc SPIE 2013 © LLTECH 2011 14
- 15. BRAIN AND SPINAL CORD © LLTECH 2011 15
- 16. 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. Cortex Neuron cell bodies (dark dots) Axon bundles Axons/myelin fibers Capillary White maHer Harms et al Proc 2011 2013 © LLTECH SPIE 16
- 17. Hippocampus: Coronal sec>on (led side) Sub-‐ Ependymal zone Pyramidal neurons of the CA4 field responsible for memory CA4 field Alveus Stratum radiatus Hippocampal sulcus of the CA1 field Stratum granulosum of the dentate gyrus CA1 field of the cornus ammonis © LLTECH 2011 17
- 18. FF-‐OCT images dis>nguish meningiomas from meningeal haemangiopericytoma Meningioma psammoma – diagnosis possible directly on FF-‐OCT image Collagen bundles Whorls CalcificaIon 80% of meningiomas are benign. Collagen balls 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. 18 © LLTECH 2011
- 19. RAT BRAIN : Different zoom levels 16x13mm2 4x4mm2 1x1mm2 Advanced Imaging in Bio © LLTECH 2011 19
- 20. Spinal cord : normal Issue 100 µm Arrows: Fibers transverse cut 500 µm 100 µm © LLTECH 2011 20
- 21. DERMATOLOGY AND COSMETICS © LLTECH 2011 21
- 22. Normal skin morphology a Epidermis c b Collagen d e 500 µm Pilosebaceous unit Sweat gland Adipocytes Dalimier and Salomon Dermatology 224, 84-92 (2012) © LLTech 2012 © LLTECH 2011 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland 22
- 23. Skin verIcal reconstrucIon from en-‐face slicing for layers assesment and measurement KN: KeraInocyte nuclei Papillary melanin caps BV: Blood vessels Stratum corneum KN KN KN Stratum spinosum BV BV BV Dermis 50 µm VerIcal reconstrucIon of skin model KeraInocyte nuclei Stratum corneum Stratum spinosum Dermis 50 µm © LLTech 2012 © LLTECH 2011 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland 23
- 24. 3D reconstrucIon from en-‐face slicing Epidermis and dermis separated by sodium bromide Epidermis Dermis 800 µm x 800 µm 800 µm x 800 µm Wrinkle Stratum corneum Stratum spinosum Dermal papilla Collagen Blood vessel 50 µm 50 µm Reconstructed verIcal slice Reconstructed verIcal slice © LLTech 2012 © LLTECH 2011 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland 24
- 25. Adipocytes 200 µm 9um depth 27um depth 60um depth Reconstructed depth slice 50 µm © LLTECH 2011 25
- 26. Skin pathologies: basal cell carcinoma discriminaIon at structural level Dense peritumoral stroma © LLTech 2012 © LLTECH 2011 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland 26
- 27. Zoom on basal cell carcinoma: discriminaIon at cellular level Peritumoral stroma High cell density 200 µm 100 µm © LLTech 2012 © LLTECH 2011 Courtesy of Hopitaux Universitaires de Genève, Genève, Switzerland 27
- 28. Skin in-‐vivo: fingerprint imaging in 3D Fingerprints 3D reconstrucIon FOV 800 µm x 800 µm Sweat ducts Corneocytes LightCT arm support add-‐ on allows in vivo imaging 100 µm on arm, hand, fingers En-‐face image of fingerprints © LLTECH 2011 28
- 29. 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 © LLTECH 2011 29
- 30. Mouse skin in vivo 20um depth En-‐face slices in thigh region 10um depth FOV 1mm2 PenetraIon to 200um Hairs near surface 35um depth 85um depth Hair follicles Collagen fibers © LLTech 2012 © LLTECH 2011 Courtesy of InsItut Langevin, Paris, France 30
- 31. LUNG © LLTECH 2011 31
- 32. Cross secIon of rat lung Scale bar =J 0.5 mm nform 2011, 2:28. Jain et al. Pathol I © LLTECH 2011 32
- 33. Image from non-‐neoplasIc human lung Issue Collapsed lung Alveoli Blood vessel FOV: FFOCT ~ 4 mm x 4 mm; H&E = 4X Jain et al. Pathology Visions 2013. © LLTECH 2011 33
- 34. Adenocarcinoma of lung with lepidic & invasive components Invasive Invasive Lepidic Lepidic Invasive Invasive © LLTECH 2011 FOCT ~ 4 mm x 4 mm; H&E = 4X . Insets: FFOCT ~ 0.375mm x 0.375mm; H&E = 20X FOV: F 34
- 35. KIDNEY © LLTECH 2011 35
- 36. Glomerulus Image from non-‐neoplasIc kidney Issue Glomerulus Blood vessel 3xzoom (FOV ~0.19x0.19 mm) Tubules Tubules 3x zoom (FOV ~0.19x0.19 mm) Blood vessel FFOCT: FOV~ 0.37x0.25 mm, depth of imaging 0.1um Caron et al. Proc SPIE 2013 © LLTECH 2011 3x zoom (FOV ~0.19x0.19 mm) 36
- 37. H&E image from non-‐neoplasIc kidney Glomerulus Blood vessel Glomerulus 20X Tubules Tubules 20X H&E = 10X Caron et al. Proc SPIE 2013 © LLTECH 2011 Blood vessel 20X 37
- 38. NeoplasIc kidney: normal architecture replaced by sheets of cells Sheets of cells Sheets of cells Sheets of cells FFOCT: FOV ~ H&E 10X 0.38x0.38 mm, depth of imaging : 7.2um 3x zoom (FOV~ 0.19x0.19mm) © LLTECH 2011 Chromophobe RCC 20X 38
- 39. RETINA AND CORNEA © LLTECH 2011 39
- 40. ReIna Photoreceptors - pig Nerve fiber layer -‐ rat OpIc Nerve -‐ pig Retinal pigment epithelium - pig ReIna -‐ rat Grieve et al IOVS 2004 Nov;45(11):4126-31. © LLTech 2012 © LLTECH 2011 Courtesy of ESPCI, Paris, France 40
- 41. Human graa cornea: over 500um thickness 50 um Ghouali et al, ARVO 2013 www.lltechimaging.com © LLTECH 2011 Courtesy of Vincent Borderie 41
- 42. Large field en face view of epithelial layer 100 um LightCT can evaluate integrity of epithelium and stroma in graa corneas (not possible with exisIng techniques) 200 um www.lltechimaging.com © LLTECH 2011 Courtesy of Vincent Borderie 42
- 43. Large field en face view in stroma 200 um ApplcaIon of LightCT in eye banks www.lltechimaging.com © LLTECH 2011 Courtesy of Vincent Borderie 43
- 44. LightCT shows corneal pathology and can penetrate even edematous cornea Cornea with edema Cornea with chemical burn – highly sca]ering upper stroma © LLTECH 2011 44
- 45. CORE NEEDLE BIOPSIES © LLTECH 2011 45
- 46. Core Needle Biopsy: Kidney LightCT provides an evaluaIon of Issue architecture in minutes 100µm Renal tubules 500µm 50µm Vessel © LLTech 2012 © LLTECH 2011 Courtesy of InsItut Curie, Paris, France 46
- 47. Core Needle Biopsy Breast: InfiltraIve carcinoma LightCT used intraoperaIvely on biopsy Issue provides fast, non destrucIve evaluaIon © LLTech 2012 © LLTECH 2011 Courtesy of InsItut Curie, Paris, France 47
- 48. DEVELOPMENTAL BIOLOGY © LLTECH 2011 48
- 49. Developmental biology: Drosophila 200 µm © LLTech 2012 © LLTECH 2011 Courtesy of ESPCI, Paris, France 49
- 50. 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 © LLTECH 2011 Courtesy of ESPCI, Paris, France 50
- 51. 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 © LLTECH 2011 51
- 52. Musée d’Histoire Naturelle Xenopus Laevis CarIlage Pigmented cells OIc vesicle 100 um © LLTECH 2011 52
- 53. Musée d’Histoire Naturelle Xenopus Laevis 50 um Reconstructed depth slices © LLTECH 2011 53
- 54. Xenopus Laevis eye: en face slices Musée d’Histoire Naturelle Surface 10um depth 20um depth 30um depth 40um depth 50um depth 60um depth 70um depth 80um depth 140um depth 230um depth 180um depth 200 um © LLTECH 2011 54
- 55. C. elegans – Images at higher magnificaIon: 40X 100 µm 3D reconstrucIon 50 µm © LLTech 2012 © LLTECH 2011 Courtesy of ENS, Paris, France 55
- 56. PLANTS © LLTECH 2011 56
- 57. Leaf veins – fresh leaf sample Veins 500 µm Fibers 100 µm 57 © LLTech 2012 © LLTECH 2011 Courtesy of ESPCI, Paris, France 57
- 58. Apple Wax Skin Water Flesh 500 µm 100 µm 58 © LLTech 2012 © LLTECH 2011 Courtesy of ESPCI, Paris, France 58
- 59. Sorghum: cross secIon Courtesy of Plate-forme d'Histocytologie et d'Imagerie Wide field en face Cellulaire Végétale (PHIV) Montpellier RIO2011 © LLTECH Imaging 14mmX15mm 59
- 60. 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 © LLTECH 2011 60
- 61. Courtesy of Plate-forme d'Histocytologie et d'Imagerie Cellulaire Végétale (PHIV) Rice: hypocotyle Montpellier RIO Imaging Wide field en face 5mmX2mm Reconstructed depth slice, 200um pentraIon depth © LLTECH 2011 61
- 62. 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 © LLTECH 2011 62
- 63. LLTech LLTech, Inc. LLTech 103 Carnegie Center Drive Pépinière Paris Santé Cochin Suite 300 29 rue du Faubourg Saint Jacques Princeton, NJ 08540, USA 75014 Paris Phone: +1 609 995 3506 Phone: +33 1 82 72 61 25 www.lltechimaging.com contact@lltech.fr www.lltechimaging.com © LLTECH 2011 63