OPTICAL COHERENCE TOMOGRAPHY FOR PATHOLOGY: FULL-FIELD OPTICAL COHERENCE MICROSCOPY

Transcription

1 OPTICAL COHERENCE TOMOGRAPHY FOR PATHOLOGY: FULL-FIELD OPTICAL COHERENCE MICROSCOPY Séminaire Cancéropôle - 19 Sept Fabrice Harms

2 Imaging in scattering media Impossible? Easy Difficult Courtesy S. Gigan Lab. Kastler Brossel

3 Optical Coherence Tomography = optical echography Similar to ultrasonography but using optical waves Measures amplitude & distance of backscattered light: selection of light coming from a defined plane under the surface First clinical applications: ophthalmology (retinal imaging) skin Typical resolution: 10 microns Typical resolution: 7 microns

4 Full Field Optical Coherence Tomography High-resolution OCT for the intraoperative diagnosis of biological tissue at the cellular scale White light source (halogen) Microscope objectives Sample 2D Camera PZ T Oscillating mirror (reference arm) High-resolution en face (C-scan) imaging = similar to pathology slides Usually based on a Linnik interferometer 1 micron isotropic 3D resolution Large fields 5x5mm in 2 minutes

5 Commercial FFOCT microscope Light-CT TM scanner, LLTech, France resolution: 1,5µm transverse, 1µm axial broadband white-light halogen source 70Hz max tomographic frame rate 1Mvoxels penetration depth 200µm 500µm depending on tissue 3D DICOM viewer : 3D reconstruction, MPR navigation 25mm diameter max. sample size

6 FFOCT resolution approches pathology requirements Cornea Scale bar 200 um Skin SDOCT (Spectralis) FFOCT Keratinocyte nuclei Stratum corneum Stratum spinosum 50 µm Dermis FFOCT SDOCT (Michelson Diagnostics) Breast Swept-source OCT (Thorlabs) FFOCT 1mm 1mm

7 Breast tissue Volume 1 e600254

8 Healthy tissue - microstructures 500µm 50µm 50µm 80µm 80µm 80µm Healthy fibrous tissue Duct with calcification Lobule Adipocytes Duct

9 Invasive adenocarcinoma nodular tumor Different aspect of the fibrous tissue 500µm fibrous tissue in the tumourous part Grey zones corresponding to foci of carcinomatous cells Grainy medium scattering aspect: healthy fibrous tissue LLTECH 2011

10 Ductal Carcinoma In Situ (DCIS) Grainy aspect of fibrous tissue : sign of non malignancy Enlarged ducts with : characteristics of DCIS 130 tumors 95% sensitivity

11 Identification of microstructures BREAST LOBULE DUCT DUCT WITH CALCIFICATION VESSEL SCAR 50µm 100µm 50µm 100µm ADIPOCYTES ABNORMAL ADIPOCYTES HEALTHY FIBROUS TISSUE FIBROUS TISSUE IN STROMA REACTION 100µm 100µm

12 Sentinel nodes 1st study Pathologist# 1 FFOCT Normal Invaded H&E Normal 27 3 Invaded 1 13 SENSITIVITY=93% SPECIFICITY=90% "Second look" : 100% sensitivity and 97% specificity, following adequate training cf. Frozen section analysis : 75% sensitivity, 100% specificity (i.e. zero false positives). A. Ali et al World Journal of Surgical Oncology 2008, 6:69 2nd study (to be submitted) 82 lymph nodes Sensitivity: 88% and Specificity : 87% Second look (after training): Sensitivity: 100% and Specificity: 93%

13 Normal sentinel node Nodules containing lymphoid tissue Adipocytes in hilum Capsule

14 Invaded sentinel node Invaded zone Lymphoid zone Bright white aspect, highly scattering dense collagen meshwork

15 Invaded sentinel node Fibrosis in hilum

16 Core needle biopsies Objective: to evaluate the feasibility of using full-field OCT (FFOCT) for immediate specimen quality assessment to decrease the number of nondiagnostic EUS-guided FNAB/core biopsies (currently 5 to 10% are useless) 24 samples from 14 patients of gastric, pancreatic, pelvic and lymph-node formalin-fixed biopsies FFOCT images (1 µm 3D resolution) acquired and later compared with histology

17 Pancreas adenocarcinoma tumorous cell sheets Adenocarcinomas represent 80% of pancreas tumors 1 mm nuclei 100µm

18 Pancreas neuro-endocrine tumor - tumorous cell sheets 500 µm nuclei 100 µm 100 µm

19 Pancreas blood clots absence of tumoral material

20 Lymph node sarcoidosis architectural modification a c fibrosis calcification Lymphoid tissue b d

21 Prostate biopsies Turkbey B et al. (2009) Imaging techniques for prostate cancer: implications for focal therapy Nat Rev Urol doi: /nrurol

22 Prostate biopsies Objective: to evaluate the ability of FFOCT to qualify intraoperatively ultrasound-mri fusion targeted prostate biopsies, thus improving procedure accuracy Blinded analysis of 116 prostate biopsy cores by 3 pathologists with regular feedback: Learning curve measurement and predicted accuracy Definition of diagnosis criteria and 2 nd blinded reading for evaluation of diagnosis performance Sensitivity 86% Specificity 88% F. Beuvon et al. (2014), Tomographie par cohérence optique plein champ des biopsies de la prostate : un pas vers le diagnostic pré-histologique? Progrès en urologie 24, Multicentric clinical study (3 centres) will start in 2015, on a significant number of samples (1800 samples, 150 patients)

23 Prostate biopsies FFOCT imaging

24 Exemple of normal prostatic gland basal layer Epithelium layer 50 µm 50 µm lumen

25 Tumorous glands 200 µm Small aggregated glands Single cell layer No or limited lumen

26 Criterion 1 - Small aggregated glands indicate a tumorous lesion Tumorous glands Benign glands

27 Criterion 2 Straight borders indicate a tumorous lesion (hard tissue =>elasticity)

28 Head & Neck tissue Objective: build a comparative FFOCT/histology atlas on Head & Neck tissue to: - evaluate the diagnosis performance of FFOCT - define training data and reading methods for pathologists 50 patients, 55 tumor samples, 67 non-tumor samples Definition of diagnosis criteria First step toward in-vivo diagnosis using FFOCT endoscopy This project has received funding from the European Union Seventh Framework Program FP7-ICT under grant agreement number

29 Example: head & neck biopsy Glossectomy, Healthy tongue tissue, 10x10mm image, 50µm depth Courtesy of Institut Gustave Roussy, Villejuif, France

30 Example: head & neck biopsy Glossectomy, Healthy tongue tissue, 10x10mm image, 50µm depth Spinous cells with visible nuclei Filiform papilla 1 mm Courtesy of Institut Gustave Roussy, Villejuif, France

31 Example: head & neck biopsy Epiglottis, healthy tissue, 8x8mm image, 50µm depth Epithelium 1 mm Cartilage Courtesy of Institut Gustave Roussy, Villejuif, France

32 Example: head & neck biopsy Epiglottis, healthy tissue, 8x8mm image, 50µm depth Cartilage 1 mm Adipocytes Epithelium Cell nuclei Courtesy of Institut Gustave Roussy, Villejuif, France

33 FFOCT allows fast non-destructive tissue processing and pathology assessment. It completes the tools available for pathologists or interventional radiologists

34 Summary key applications

35 Summary key features FFOCT shows good performance in clinical contexts where a fast, nondestructive tissue assessment is required: it can supplement existing tools to better select informative or adequate tissue, thus reducing repeated surgery or biopsy Many more images of various types are available (Brain, Lung, Cornea grafts, kidney, pancreas, etc.), with good and bad points (e.g. false positive) One particularly challenging task: to positively interact with clinicians: Define new reading and diagnosis criteria (forget H&E reflexes) Find time! (this is an iterative work)

36 Current developments & future devices Development of a medical device for fast, non-invasive intraoperative assessment of biopsies Additional contrast modalities for the improvement of diagnosis accuracy: Elastography Polarization Fluorescence Etc.?

37 In-vivo biopsy & surgery guidance => FFOCT endoscopy ex-vivo breast tissue Human skin (forearm), normal and mole tissue Stratum Spinosum, depth 20 µm This project has received funding from the European Union Seventh Framework Program FP7-ICT under grant agreement number

38 In-vivo biopsy & surgery guidance => FFOCT endoscopy Custom high-performance CMOS sensor Custom high-speed CMOS camera Clinical cellular resolution imaging devices Clinical assessment High speed biopsy imaging non-invasive endomicroscopy -Breast tumor margins -Head & Neck surgery guidance This project has received funding from the European Union Seventh Framework Program FP7-ICT under grant agreement number

39 ADOC: Intraoperative digital diagnosis in cancer surgery General objective: To develop a device for intraoperative diagnosis in cancer surgery, based on FFOCT, including fast, automated retrieval of structural characteristics of tissue at a cellular level. To indicate the surgeon wether to continue or not removing tissue within a few minutes. Medical imaging Software Medical image processing Non-invasive highresolution optical imaging Medical device for non-invasive biopsy assessment with automated morphology analysis Clinical expertise in cancerology

40 THANK YOU Drosophila Core needle biopsy - kidney C Elegans Skin in Skin - BCC C Elegans