• Histopathologic examination involves making tissue • Appendix: Submit a cross-section of the base and diagnoses, taking into account the clinical and/or middle, and a longitudinal section of the tip. radiographic data, to help clinicians manage patients
SAFETY PRECAUTIONS WHEN HANDLING
SPECIMENS • Wear personal protective equipment (e.g., face mask, gloves, apron) and treat all tissues as potentially infectious. Additionally, specimens are submerged in 10% neutral buffered formalin which may cause irritation of the skin and airways. • Clean used materials and cutting area after use. Wash hands properly after handling specimens
GENERAL APPROACH TO SPECIMEN DISSECTION
• Identify the specimen submitted. Cross check patient details written on the container and on the requisition form. • Gross description: Describe the consistency (e.g., soft, doughy, hard), shape, and color of the tissue. Specimens such as gallbladders may be sectioned/ opened prior to submission; this should be reported in • Gallbladder: Submit a cross-section of the neck, the the gross description. body, and the fundus. • Some tissues (e.g., prostatic chips, uterus) must be weighed. . • Measure the specimen in 3 dimensions (L x W x H). Tubular structures such as the appendix, fallopian tubes require only 2 measurements (L x cross- sectional diameter). • If the specimen requires orientation (e.g., breast masses that require reporting of margins), ensure proper orientation prior to sectioning. Mark the margins with ink. • Specimens are generally sectioned in a cross- sectional fashion unless otherwise indicated (e.g., the tip of the appendix, bowels) • Once the specimen is sectioned, describe its cut surface and submit tissue sections.
OVERVIEW OF TISSUE PROCESSING
• Tissue sections are submitted in properly labeled tissue cassettes. The cassettes are then sequentially • Endometrial curettings: Measure the aggregate immersed in different reagents in a process that takes diameter of the tissue fragments. For cases of at least 12 hours abnormal uterine bleeding, submit all tissues. • This process involves dehydration (using ethanol), • Core needle biopsy specimens: Measure the length clearing (using reagents such as toluene, xylene), of the strips and wrap in a piece of filter paper to and infiltration (with paraffin). Afterwards, the tissues ensure the specimen stays in the tissue cassette. are embedded into molten wax to create tissue blocks • Breast mass (e.g., lumpectomy): Ink the exterior from which sections will be obtained surface then serially cut (“bread loaf”) the tissue and • Thin sections are cut from the blocks using a submit representative sections of the mass. microtome and placed on slides for staining. • Hematoxylin and eosin stains are used for routine IMMUNOHISTOCHEMICAL STAINS sections. However, special stains such as Giemsa, • Tumors may exhibit different morphologies and also Periodic Acid Schiff, etc may be used to highlight mimic the appearance of other tumors. If a definitive certain cells/ features of tissues. diagnosis cannot be made using H&E stained sections, immunohistochemical stains may be used to help clinch the diagnosis. • Immunohistochemical stains or immunostaining uses antibodies complexed with dyes (commonly brown) to detect specific proteins in tissue sections. • In other cases, these stains are used to further evaluate the characteristics of tumors. For example, the commonly requested stains for Estrogen receptors (ER), Progesterone receptors (PR), and HER2/neu in cases of breast cancer o These are interpreted based on the intensity of staining of individual cells and the proportion of cells that take up the stain. o The ER/PR/HER2neu status is important for the post-surgical treatment of patients
1 ANATOMIC PATHOLOGY
FINE NEEDLE ASPIRATION BIOPSY
• It is used to describe the technique of aspirating cells and tissue fragments through a thin needle 22- 27 gauge (up to 0.8 mm OD). Aspiration can be achieved using the capillary pressure of the needle itself or by applying a partial vacuum using a syringe. • Diagnostic results can be reported in a manner similar to other types of cytological examination: (1) unsatisfactory (inadequate), (2) negative for malignancy (benign), (3) atypical, probably benign (atypical/undetermined), (4) suspicious of malignancy (probably malignant), and (5) positive for malignancy (malignant).
FNAB TECHNIQUES 1. Depending on the clinical situation, the cytopathologist, surgeon, or radiologist may perform the FNA. The FNA procedure should be clearly explained to the patient to assure patient’s cooperation during FNA procedure. 2. The patient should be placed in a comfortable position that allows easy access to the lesion that needs to be aspirated. 3. For FNA of palpable, superficial lesions, the skin over the area of the procedure is cleaned with an alcohol or with another antiseptic solution. 4. For palpable lesions a 27- to 22-gauge (0.4–0.7 mm) needles are suitable, either using a capillary technique without aspiration or a plastic disposable 10- or 20-ml syringe attached to a plastic or metal syringe holder (FNA with aspiration) 5. For guided FNAs of non-palpable, deeper lesions, longer needles are utilized. 6. The nodule is immobilized between the fingers, and the needle tip is rapidly directed through the skin into the nodule. 7. Once the needle enters the mass, the needle is continuously aspirated, while the needle is rapidly moved back and forth to obtain the sample. 8. Suction is then relieved and the needle is withdrawn and detached from the syringe. 9. Air is then aspirated into the syringe, the needle is replaced onto the syringe, and the material is expelled from the needle onto the glass slides. 10. The material is gently but rapidly smeared on the slides and immediately dipped in fixative. For liquid based preparation the aspirate is collected directly in specially developed liquid fixative.
2 ANATOMIC PATHOLOGY CONVENTIONAL PREPARATION OF FNA • The principle is to spread a suitable amount of aspirate (usually about one drop) thinly and evenly over a microscopic slide that is then stained and mounted. • Two common methods of fixation: 1. AIR DRYING can be stained with Diff-Quik or May-Grünwald-Giemsa (MGG) give better information on cytoplasmic details (see Fig. 1.17), as well as the extracellular matrix (see Fig. 1.18) and the background material.
2. WET FIXING USING 95% ETHANOL
can be stained with hematoxylin and eosin (H&E) or Papanicolaou (Pap) better demonstrates such details as nuclear pattern, chromatin structure, and nucleoli
3 ANATOMIC PATHOLOGY CELL BLOCKS (CB) Smears can be difficult to prepare reliably as a result of the nature of the aspirate (cystic components, blood contamination) or due to inexperience in the person making the smear. Of the various techniques devised to make best use of aspirates, the preparation of cell blocks (CB) can be very helpful. CB preparation advantages are twofold: (1) better visualization of the tumor tissue pattern than is possible in FNA smear (see Figs. 1.23, 1.24, and 1.25) and (2) an opportunity of performing immunocytochemical and other special stains on several slides of comparable quality.
IMMUNOCYTOCHEMISTRY Designed to improve routine cytology to aid pathologist/cytopathologist in the differential diagnosis of neoplasm and to render confident and accurate diagnoses on limited tissue samples. May be applied to any of the several types of preparations: to direct smears, cell-transferred direct smears, cytospin preparations, liquid-based preparations, or cell blocks. Cell block preparations using paraffin-embedded cells are becoming increasingly popular in many busy FNA clinics, allowing both examination of the tissue architecture and making immunostaining more reliable.
REFERENCES • William H. Westra, M.D., et al. Surgical Pathology Dissection: An Illustrated Guide, Second Edition • Henryk A. Domanski, Atlas of Fine Needle Aspiration Cytology, 2nd edition
Get (Ebook) Orell and Sterrett's Fine Needle Aspiration Cytology, 5th Edition by Svante R Orell, Gregory F. Sterrett MB BS FRCPA FIAC ISBN 9780702031519, 0702031518 PDF ebook with Full Chapters Now
Instant Download (Ebook) Fine Needle Aspiration Cytology: Interpretation and Diagnostic Difficulties by Pranab Dey ISBN 9789385999383, 9385999389 PDF All Chapters
Get (Ebook) Orell and Sterrett's Fine Needle Aspiration Cytology, 5th Edition by Svante R Orell, Gregory F. Sterrett MB BS FRCPA FIAC ISBN 9780702031519, 0702031518 PDF ebook with Full Chapters Now
Instant Download (Ebook) Fine Needle Aspiration Cytology: Interpretation and Diagnostic Difficulties by Pranab Dey ISBN 9789385999383, 9385999389 PDF All Chapters
