Schedule of Accreditation issued by United Kingdom Accreditation Service 2 Pine Trees, Chertsey Lane, Staines-upon-Thames, TW18 3HR, UK

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1 2 Pine Trees, Chertsey Lane, Staines-upon-Thames, TW18 3HR, UK Great Ormond Street Hospital for Children NHS Foundation Trust Department of Histopathology Floor 3 Camelia Botnar Laboratories, Great Ormond Street Hospital for Children NHS Trust, Great Ormond Street, London WC1N 3JH Contact: Dr. Michael Ashworth Tel: +44 (0) Website: www. gosh.co.uk Testing performed at the above address only Additional site activity performed away from the laboratory Location details Mortuary and Body Store Floor 0 Camelia Botnar Laboratories Great Ormond Street Hospital for Children Great Ormond Street London WC1N 3JH Activity Storage of Bodies Record Keeping (body identification and audit trail) Fresh tissue DETAIL OF ACCREDITATION Histopathology examination diagnosis Specimen Dissection SSOP SSOP Tissue processing Leica, Peloris and Sakura, VIP in procedures: SSOP SSOP 01.11a SSOP Assessment Manager: SGB Page 1 of 17

2 Formalin fixed tissue Histopathology examination diagnosis Routine H and E staining for tissue architecture and nuclear detail (identification of basophilic and eosinophilic structures) Tissue embedding Sakura, Tissue Tek in conjunction with documented procedures: SSOP Tissue sectioning Leica, RM2245 and Thermo Fisher Shandon, ME+ in conjunction with documented procedures SSOP FFPE Leica CV 5030 Autostainer in procedures: SSOP SSOP SSOP SSOP SSOP SSOP SSOP SSOP Fresh Tissues Frozen Section Leica C 1860 cryostat and Thermo Electron Cryotome in conjunction with documented method: SSOP Assessment Manager: SGB Page 2 of 17

3 Fresh Tissue /Frozen Section Histopathology examination for the purpose of diagnosis Special Staining to identify or exclude morphological Eosinophilic and basophilic structures Fats and lipids Lipid Complement Manual staining methods using in house documented procedures and the following stains: SSOP 05.41: Rapid Haematoxylin and Eosin (H&E) for frozen section and cytospin Oil red O SSOP Sudan Black SSOP C4d SSOP05.41 Fresh tissue smear Differentiation of myopathies Fresh Tissue smear (Tol Blue) SSOP12.04: Special staining Examination of tissues to identify or exclude morphological and cytological abnormalities Weakly sulphated and acidic mucins Acidic and neutral mucins Amyloid Elastic connective tissue fibres Manual staining methods using in house documented procedures and the following stains: Alcian Blue (AB) technique SSOP Alcian Blue Periodic Acid-Schiff (AB PAS) technique SSOP Congo Red technique SSOP Elastic van-gieson (EVG) (Miller s) technique SSOP Assessment Manager: SGB Page 3 of 17

4 Histopathology examination diagnosis Special staining to identify or exclude morphological Haematopoietic cells and Helicobacter pylori-like organisms Manual staining techniques using in house documented procedures and the following stains: Giemsa technique SSOP Myofibrils Gomori Trichrome SSOP Elastic fibres, mast cells, suphated mucins Gomori Aldehyde Fuchsin SSOP Detection of organisms Gram stain SSOP Agryophil cells Grimelius SSOP Detection of fungi Grocott SSOP Basophilic and eosinophilic structures Urine mucopolysaccharide Haematoxylin and Eosin SSOP and 41 Haust and Landing Toluidine Blue SSOP Mucins High Iron Diamine SSOP Carbohydrate Lugols Iodine SSOP Myelin sheaths Luxol Fast Blue SSOP Fibrin Martius Scarlet Blue (MSB) SSOP Melanin and argetaffin granules Masson Fontana SSOP Collagen Masson one step SSOP Connective tissue elements Masson Trichrome technique SSOP Assessment Manager: SGB Page 4 of 17

5 Histopathology examination for the purpose of diagnosis Special staining to identify or exclude morphological Nucleic acids, polysaccharides Manual staining techniques using in house documented procedures and the following stains: Methanolic Toluidine Blue SSOP Nuclear material Nuclear Fast Red SSOP Hepatitis B surface antigen Orcein SSOP Glycogen, mucopolysaccharide components, fungi and basement membranes Pituitary cell types Periodic Acid Schiff SSOP Periodic Acid-Schiff (PAS) Orange G SSOP Polysaccharides Periodic Acid Schiff Diastase Celloidin Glycogen Renal pathology (BM) Periodic Acid Schiff-Celloidin SSOP Periodic Acid Methanamine SSOP Ferric Iron Perls Prussian Blue SSOP Copper Rhodanine SSOP Melanin Schmorl SSOP Mast cells / metachromatic tissue components Toluidine Blue technique SSOP 11.24/35/36 Calcium Tripp and Mackay SSOP Collagen connective tissue fibres van-gieson (HVG) SSOP Assessment Manager: SGB Page 5 of 17

6 Histopathology examination for the purpose of diagnosis Special staining to identify or exclude morphological Manual staining techniques using in house documented procedures and the following stains: Calcium Von Kossa SSOP Mycobacterium e.g. M. leprae Wade Fite SSOP Infectious disease eg spirochetes Warthin Starry SSOP and fresh frozen tissue on Acid and alcohol-fast bacilli Histopathology examination for the purpose of diagnosis Immunohistochemistry. Identification of : Adenovirus Cytokeratin (CK), LMW CK (LMW & HMW) Fusion protein eg lymphomas Amyloid precursor Histiocytes/Hepatocellular Ca Hepatic tumours Embryonic cell in normal tissue and germ tumours Basement membrane Astrovirus Muscle fibres Ziehl-Neelsen (ZN) technique SSOP Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP Adenovirus AE1 AE1/AE3 ALK-1 Alzheimer precursor Alpha 1 Antichymotrypsin Alpha 1 Antitrypsin Alpha Feto-protein Alpha V Integrin Astrovirus B-1 Integrin Assessment Manager: SGB Page 6 of 17

7 and fresh frozen tissue on Histopathology examination for the purpose of diagnosis Immunohistochemistry Identification of : Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP Carcinoma Beta-catenin Lymphoma/Carcinoma Bcl-2 Lymphomas Bcl-6 Lymphoma BerH2 Epithelial cell types Ber-EP4 Tumour marker eg Carcinoma Bombesin Complement (Renal disease) C1q Complement (Renal disease) C3 Gastrointestinal and pulmonary Calcitonin disease Ganglion cells (Hirschsprung Calretinin disease) Apoptosis Caspase 8 Apoptosis Caspase 3 -Cleaved Cardiovascular disease Caveolin Cardiovascular disease CCC9 T antigen on lymphocytes CD3 T cell subset CD4 T cell subset CD8 T cell marker CD20 Endothelial marker CD31 Carcinoma/Lymphoma CD10 Histiocytes/Langerhan cell CD1a GIST,RCC,Germ cell tumour CD117 (c-kit) Stem cells and Neuro tum. CD133 Lymphoma CD138 Histiocytes CD14 Lymphoma CD15 Monocyte/macrophage cells CD163 Pan t-lymphocytes CD2 Pan B-cell CD20 Folicular Dendritic cells CD21 Lymphomas CD25 Assessment Manager: SGB Page 7 of 17

8 and fresh frozen tissue on Histopathology examination for the purpose of diagnosis Immunohistochemistry Identification of : Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP Lymphomas CD30 Stem cell/endothelium sarcoma CD31 Stem cell/endothelium sarcoma CD34 Lymphoma CD4 Neuropath, glial cells CD44 Pan Leukocyte marker (T&B) CD45 Pan T-cell marker CD5 Neural cell adhesion molecule CD56 NK cells, neuroendocrine cells CD57 Pan B-cell marker CD79a Lymphoma CD8 Sarcomas, spindle cell tumours CD99 Carcinoma CEA (Carcino Embryonic Antigen) Neuroendocrine Chromogranin Cytomegalovirus CMV Cytomegalovirus CMV in situ Cytokeratin in specific epithelium Cytokeratin 7 (CK7) CK for Carcinoma Cytokeratin 14 (CK14) CK for Carcinoma Cytokeratin 19 (CK19) CK in gastrointestinal cells Cytokeratin 20 (CK20) Cellular proliferation/differentiation C-MYC and neoplastic disease Collagen (Neuromuscular dis.) Collagen 6 Lymphatic endothelium D2-40 Intermediate filaments Desmin D33 Epstein Barr Virus EBV Epstein Barr Virus EBV-ISH Lymphoma/carcinoma/meningioma Epithelial Membrane Antigen Cardiac and skeletal muscle Emerin Endothelial cells Factor VIII Histiocytic and Connective tissue Factor 13a tumours Lymphoma (Hodgkins) Fascin Assessment Manager: SGB Page 8 of 17

9 and fresh frozen tissue on Histopathology examination diagnosis Immunohistochemistry Identification of : T cell marker squamous carcinoma Leukaemia Neuroendocrine cell Human chorionic gonadotrophin Human Growth Hormone Lymphocytes Melanocytes (melanoma) Herpes Simplex I/II Herpes Simplex I Herpes Simplex II Carcinoma vs Germ cell tumour Tumours (rhabdoid, sarcoma) Endocrine Macrophages, microglia Leucocytes Immunoglobulin A Immunoglobulin G Immunoglobulin M Interleukin 8 Kappa light chains Kappa and Lambda light chains Mitosis Macrophages Lambda light chains Lysosomal associated membrane protein Lysosomal associated membrane protein Langerhans cells Luteinizing Hormone Macrophages Endothelial cell marker Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP Foxp3 GLUT-1 Glycophorin C Glucagon HCG Human Growth Hormone HLA DR HMB45 Herpes Simplex I and II Herpes Simplex I Herpes Simplex II Inhibin INI-1 Insulin Integrin AV1 Integrin B1 IgA IgG IgM IL-8 Kappa Kappa and Lambda in situ Ki67 KP1 Lambda Lamp-1 Lamp-2 Langerin LH Lysozyme LYVE-1 Assessment Manager: SGB Page 9 of 17

10 and fresh frozen tissue on Histopathology examination diagnosis Immunohistochemistry Identification of : Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP Macrophage marker MAC 387 Microtubule marker MAP2 Mast Cells Mast Cell Tryptase Muscle (neuropathology) Merosin Sarcoma/Lymphoma MIC2 Cytokeratin (Broad Range) MNF 116 lympho-haemopoietic neoplasms MUM-1 Leukaemia Myeloperoxidase Small Round cell tumours NB84 Germ cell tumours OCT-3/4 Endothelium O-Nectin Ganglion cells/macrophages O-Pontin Lymphoma/neuropath P4EBP Tumour marker P53 Tumour marker P62 CNS marker Parvalbumin Lymphoma PAX5 Neuroendocrine Pancreatic cocktail Tumour marker P component Macrophage marker PGM1 Tumour marker Plakoglobin Tumour marker PLAP Pancreatic polypeptide Pancreatic polypeptide Prealbumin Prealbumin Proinsulin Proinsulin Prolactin Prolactin Schwann cells, nerve processes, S100 S100 Positive neoplasms Lung Surfactant B Tumour (eg RCC) TFE3 Thyroid Thyroglobulin Toxoplasma Toxoplasma Prealbumin, Liver Transthyretin Assessment Manager: SGB Page 10 of 17

11 and fresh frozen tissue on Neuromuscular tissue samples Histopathology examination diagnosis Immunohistochemistry Identification of : Growth factor Mesenchymal cell origin Tumour differentiator Tumour marker e.g. Wilms Neuropathology examination diagnosis Leica Bond Max Immunohistochemistry in procedures and the following antibodies: SSOP VEGF VG1 Vimentin V-CAM WT-1 Specimen Dissection SOP12.01 SOP12.04 SOP1223 Tissue processing Leica, Peloris and Sakura, VIP in procedures: SOP01.11 SOP01.11a SOP01.15 Embedding Sakura, Tissue Tek in conjunction with documented procedures: SOP01.16 Assessment Manager: SGB Page 11 of 17

12 Neuromuscular tissue samples and fresh frozen tissue on Brain Smear CSF Neuropathology examination diagnosis Neuropathology examination diagnosis Neuropathology special staining (automated) Tissue sectioning Leica, RM2245 and Thermo Fisher Shandon, ME+ in conjunction with documented procedures SOP01.17 (FFPE) SOP02.17 (Frozen) Leica CV 5030 Autostainer in procedures and the following special stains: SSOP SSOP12.04 SSOP03.01 Neurofibrillary tangles Bielschowsky SSOP Nucleic acids NISSL SSOP Myelin sheaths (PNS) Solochrome Cyanin SSOP Nerve fibres Glees and Marsland SOP Myelin sheaths (CNS) Luxol Fast Blue (Kluver and Bareera) SSOP Tumour marker eg Neuropath Brachyury Neuronal marker Calbindin Pituitary FSH Neuropath (growth factor) GAB Neuropath (polyglutamine 1C2 expansion) Neuropath (Glial tumours) IDH Neuropathology (neural stem cells) Nestin Neuronal cell types (neuropath) Neu N Neuronal cells Neurofilament Assessment Manager: SGB Page 12 of 17

13 and fresh frozen tissue on Neuropathology examination diagnosis Neuropathology histochemical staining (automated) Leica CV 5030 Autostainer in procedures using the following histochemical stains: Neuronal cells Neurofilament 200KD Neuronal Cells NSE Neurons, melancytes, PGP 9.5 neuroendocrine cells Neuropathology PS6 Neuropathology RT97 endocrine dysfunction Somatostatin Purkinje cells Spectrin Thyroid stimulating hormone TSH Neuronal staining TUJ-1 Classification of dementias Ubiquitin Classification of medulloblastomas YAP Muscle Muscle (neuropath) Dynorphin Muscle Dystrophin Dystrophin 1 Muscle Dystrophin Dystrophin 2 Muscle Dystrophin Dystrophin 3 Limb girdle Muscle dystrophies Dysferlin CNS tumours (eg. ETMR) LIN-28a Rare muscle dystrophies Alpha dystroglycan Rare muscle dystrophies Beta Dystroglycan Myoglobin Myoglobin sub typing muscle fibres Myosin -Developmental sub typing muscle fibres Myosin Fast Skeletal muscle Myogenin sub typing muscle fibres Myosin Neonatal sub typing muscle fibres Myosin -Slow Mac-C5b-9 Mac-C5b-9 Sarcoglycanopathies Sarcoglycan Alpha Sarcoglycanopathies Sarcoglycan Beta Sarcoglycanopathies Sarcoglycan Delta Sarcoglycanopathies Sarcoglycan Gamma Actin Smooth Muscle Actin Smooth Muscle Assessment Manager: SGB Page 13 of 17

14 Neuropathology examination diagnosis Neuromuscular Enzyme Histochemistry (manual) In house documented manual methods using the following enzymes: SSOP 10.01/02/03/05/06 SSOP SSOP Acetylcholinesterase Acetylcholinesterase SSOP Acetylcholinesterase Rapid Acetylcholinesterase SSOP Acid Phospatase Acid Phosphatase Gomori SSOP Acid Phosphatase Acid Phosphatase AS-BI SSOP Adenylate Deaminase Adenlyate Deaminase SSOP Alcohol Dehydrogenase Alcohol Dehydrogenase SSOP Alkaline Phosphatase Alkaline Phosphatase BCIP-NBT SSOP Alkaline Phosphatase Alkaline Phosphatase Napthol AS-TR SSOP ATPase ATPase Fibre typing SSOP Alpha Glucosidase Brush Border Alpha Glucosidase SSOP Cystine crystals Cystine crystals SSOP Cytochrome Oxidase Cytochrome oxidase SSOP Glucose 6 Phosphate Glucose -6-Phospatase SSOP Glycerol Phosphate dehydrogenase Glycerophosphate dehydrogenase (Vit K) SSOP Lactase Lactase Azo coupling method SSOP Lactase Lactase Indigogenic method SSOP NAHD Dehydrogenase NADH Dehydrogenase SSOP Esterase Non-specific Esterase SSOP Assessment Manager: SGB Page 14 of 17

15 Neuropathology examination diagnosis Neuromuscular Enzyme Histochemistry (manual) In house documented manual methods using the following enzymes: SSOP 10.01/02/03/05/06 SSOP SSOP Nucleic acids Nuclear Fast Red SSOP Mucins, fungi and carbohydrates Periodic Acid Schiff (PAS) SSOP Phosphofructokinase Phosphofructokinase SSOP Phosphylase Phosphoylase SSOP Picrosirius Picrosirius SSOP Succinate Dehydrogenase Succinate Dehydrogenase MTT SSOP Lipids, lipoproteins Sudan Black Propylene Glycol Nuclear staining SSOP Toluidine Blue ( Haust and Landing) SSOP Nuclear staining Toluidine Blue (MLD) SSOP Nuclear staining Toluidine Blue (Meth) SSOP Muscle architecture Trichrome Gomori SSOP Assessment Manager: SGB Page 15 of 17

16 Formalin and glutaraldehyde fixed tissues Hair Platelets Buffy Coat Urine Neuropathology examination diagnosis Electron Microscopy Demonstration of ultrastructural components of renal, muscle, cardiac, nerve and other tissues Production of micrographs for diagnostic and interpretive use. Demonstration of ultrastructural components Demonstration of ultrastructural components (Hermansky Pudlak) Demonstration of ultrastructural components - Vacuolated lymphocytes Demonstration of ultrastructural components - Metachromic Granules JEOL 1400 Electron Microscope and Leica Ultracut Microtome with in house documented methods: SSOP 09.03: Specimen reception, fixation and dissection SSOP 09.04: Resin Processing SSOP 09.07: Semi thin sectioning using the Ultra microtome SSOP 09.08: Ultrathin sectioning SSOP 09.10: EM staining schedule SSOP (B): Reception and booking in and Omni Reference guide SSOP Diagnosis of Hermansky Pudlak (HP) SSOP SSOP Assessment Manager: SGB Page 16 of 17

17 : Cytopathology examination for the purpose of diagnosis examination: SSOP 03.03: Use of Cytospin SSWOP 02.01: Handling of fresh/infectious specimens and use of Hepair Class 1 cabinet SSOP 06.01: Class 1 safety cabinet operating procedure SSOP 03.07: Use of centrifuge CSF SSOP 03.10: Cerebro Spinal Fluid (CSF) Urine SSOP 03.05: Urine for metachromatic granules Body Fluids SSOP 03.01: Body Fluids Cyst Aspirates SSOP 03.11: Craniopharyngioma cyst aspirate Sputum SSOP 03.04: Sputum SSOP Oil Red O SSOP 05.41: Rapid Haematoxylin and Eosin (H&E) for frozen section and cytospin Trans bronchial Biopsy and lavage SSOP 03.03: Transbronchial Biopsy and lavage All prepared as above Morphological assessment and interpretation/diagnosis SRP002 END Assessment Manager: SGB Page 17 of 17