Kann die konfokale Mikroskopie Nachresektionen ersparen?

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1 Kann die konfokale Mikroskopie Nachresektionen ersparen? Ist der Pathologe ersetzbar?, B. Papassotiropoulos, Z. Varga, L. Moskovszky, M. Nap, U. Gueth, A. Baege, E. Amann, F. Chiesa, C. Tausch

2 In der Diagnostik und Therapie von Mammakarzinom ist die Beurteilung der Gewebeoberfläche von grosser Bedeutung.

3 Confocal laser microscopy Julien Bobroff, Laboratoire de Physique des Solides, Universite Paris-Sud

4 Histolog Scanner TM (SamanTree, Medical SA, Switzerland)

5 Entnahme der Biopsie Anfärbung mit Fluorochromen Scan Prozess Weiter nach Standard-Verfahren

6 Microfocal scanner image H&E Färbung Normalbefund Invasives Karzinom

7 Evaluation der confocal microscopy an Corebiopsien 24 Patientinnen wurden eingeschlossen 42 Bilder Zwei Pathologen führten 84 diagnostische Beurteilungen nach der B- Klassifikation durch

8 Pathology Reporting for Minimal Invasive Biopsies B1 = Normales Drüsengewebe B2 = Benigne Läsion B3 = Benigne Läsion mit unsicherem biol. Potenzial B4 = Malignitätsverdächtig B5 = Maligne B- Classification (National Coordinating Group for Breast Screening Pathology (NHSBSP), E.C. Working Group on Breast Screening Pathology, S3- Leitlinien

9 Table 1. Results (Numbers of NSC = narrow sense correspondence, BSC = broad sense correspondence, NO = mismatch, ND = could not be determined). In most cases, one H&E image- based classification was compared to two HS image-based classifications Übereinstimmung in 76 von 80 Bildvergleichen

10 Was bedeutet das für die Praxis? Die Methode ist für die schnelle Oberflächenbeurteilung von Biopsien auf Karzinomzellen hin geeignet. Wir sehen Potenzial für die intra-operative Beurteilung von Schnitträndern. Die Immunhistochemie zur Typisierung ist weiterhin essentiell.

11 Ex-vivo performance evaluation of the Histolog Scanner for human breast carcinoma detection on fresh breast core biopsies Background Bärbel Papassotiropoulos 1, Zsuzsanna Varga 2, Linda Moskovszky 2, Marius Nap 3, Constanze Elfgen 1, Astrid Baege 1, Esther Amann 1, Federica Chiesa 1, Uwe Güth 1, Christoph Tausch 1 1 Brust-Zentrum Seefeld, Zurich, Switzerland 2 Institute of Pathology and Molecular Pathology, University Hospital Zurich, Zurich, Switzerland 3 Nap Pathology Consultance bv, Numansdorp, The Netherlands Breast conserving surgery is one of the most standard operation methods for breast cancer. In order to avoid re-resections and to achieve oncological safety it is crucial to obtain tumor-free margins. Current intraoperative imaging methods as well as widely-used pathologic frozen section analysis (FSA) assist surgeons in removing the entire tumor tissue. However, limitations are well-known with regard to tumor types in case of imaging methods or time-consumption, diagnostic accuracy and freezing artefacts in case of FSA. Confocal microscopy, a high-resolution imaging method without the need of tissue freezing, may contribute to rapid and accurate intraoperative evaluation of breast specimen margins. In this study we aimed to evaluate the ability of the Histolog TM Scanner (HS) to detect breast cancer in fresh human breast core biopsy samples. Methods Equipment and Imaging: The Histolog TM Scanner (v1.0, SamanTree Medical SA, Lausanne, Switzerland, CE marking) is based on confocal fluorescence displays microscopic histology images of superficial layers of fresh tissue after nuclear staining with Acridine Orange (30 seconds) and rinsing in saline solution Fluorescence is excited by a laser diode at the wavelength of 488 nm and fluorescence emission is collected in the wavelength above 500 nm Scanning area: 16 x 16 mm high-resolution images of 8000 x 8000 pixels; preview-images with resolution of 1600 x 1600 pixels Study procedure: Twenty-four breast core biopsies were collected from lesions suspicious for breast cancer during clinical breast assessments (23 US-guided and 1 MG-guided). Immediately prior to gold standard pathology workflow (formalin fixation), Histolog TM Scanner was used to image the fresh biopsy specimens. One to two high-resolution images were taken per specimen. Each image served as an observation unit. Pathologic assessment: Two independent pathologists assessed the Histolog TM Scanner- and H&Estained images subsequently according to the B-Classification (Categories B1-B5; 0 was defined as no diagnosis possible ) and determined the correspondence of the results. Correspondence assessment: Narrow-sense correspondence (NSC): Allocation to the same category. Broad-sense correspondence (BSC): Allocation to categories that only differed with regard to pathologists confidence (B2 and B3; B4 and B5). Conclusions H&E- Images Results Pathologic results: Twenty four biopsy specimens were imaged with the Histolog TM Scanner, resulting in 42 high-resolution images and 84 pathologic assessments (two pathologists involved) 80 pathologic assessments led to diagnosis according to B-classification In 4 assessments diagnosis not possible Correspondence assessment of HS- and H&E-derived diagnoses resulted in a total of 76 correspondences (95%) NSC in 41 of 80 diagnoses (51%) and BSC in 35 of 80 diagnoses (44%) Four mismatches occurred Normal breast tissue structures: ductal and lobular structures, fat tissue and connective tissue. Final H&E-stained microscopic assessment resulted in 21 invasive carcinoma, 1 in-situ-carcinoma, 2 benign lesions Histolog TM Scanner Acquire Images (NSC) 2 2 (NSC) 5 (BSC) (ND) 1 (NO) 4 (NSC) 5 3 (ND) 2 (NO) 1 (NO) 30 (BSC) 33 (NSC) Technical results: Mean size of specimens: 11.5 mm (2 to 20 mm) Mean time to 1. HS-image (including initial preparation/30 s-staining): 6 min (range 3-9 min) Mean time to 2. HS-image (including new positioning): 3 min (range 1-6 min) Mean time to obtain diagnosis of HS-images: 12 seconds (range 5-60 s) Max. time until formalin fixation (gold standard): 10 min -> no tissue damage occurred Device easy to use, 2 hour training for medical operators is sufficient We observed a satisfying rate of correspondence between Histolog TM Scanner- and H&E-derived diagnoses on breast core biopsies. Thus, the device sufficiently detects breast cancer in fresh human tissue. In addition, Histolog TM Scanner imaging and assessment procedure was fast and breast tissue was not harmed by scanning procedure. Further studies evaluating margins of surgical excision specimens are warranted. Disclosure statement: The study was supported by SamanTree Medical SA.

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