CYTOLOGY

INTRODUCTION:- Cytopathologists are concerned about and committed to quality assurance and quality control in their laboratories. These practices include, among others, the use of intralaboratory and extradepartmental consultations, case reviews, correlation of cytologic and histopathologic specimens and review of completed diagnostic reports. QUALITY ASSURANCE MEASURES:- Cytopathology is a practice of medicine and represents a medical consultation, in both gynecologic and nongynecologic anatomic sites. The basic principles of quality assurance apply to all types of cytologic specimens. The following represents several minimum quality assurance measures. 1. Laboratory Directors:- *The laboratory should be directed by a legally qualified physician with a specialist qualification in pathology, including special training and expertise in cytopathology. *The director or designated medical professional is responsible for proper performance and reporting of all tests done in the cytopathology laboratory. *The director or designated cytopathologist should be physically present in the laboratory to direct the staff, be available for consultations, review all reactive and abnormal gynecologic cytology samples, review fine needle aspiration samples, and review all nongynecologic samples. 2. Cytotechnologists:- A suitably qualified person should be recruited for this position. 3. Physical Laboratory Facilities:- *The laboratory should be clean, well lighted, adequately ventilated, and functionally arranged so as to minimize problems in specimen handling, evaluation, and reporting. *The area for specimen preparation and handling should be separate from the area where specimens are evaluated and reported. *Formaldehyde and xylene (if in use) should be carefully monitored due to the possible presence of hazardous vapor concentrations. 4. Safety Precautions:- *Laboratory personnel must be protected against hazards (chemical, electric, fire, infections, or others) by using well-ventilated hoods and biologic safety hoods for handling potentially infectious material. * Fire precautions should be posted and tested. 5. Equipment:- *An adequate number of binocular microscopes of good quality and proper working order must be available. * Laboratory instruments and equipment should be under periodic maintenance to monitor and ensure malfunctions do not adversely affect analytical results. 6. Specimen Collection:- *Cytologic specimens should be accepted and examined only if requested by a licensed medical practitioner and collected in accordance with instructions regarding recommended collection techniques. *The cytopathology laboratory should inform the originator of the sample if the specimens are “unsatisfactory” and detail adequacy qualifiers such as presence or absence of a transformation zone component or obscuring factors in “satisfactory samples”. 7. Preparation, Fixation, and Staining Procedures:- *The specimens must be identified with the patient’s name and/ or a unique identifier and must be accompanied by a requisition form with the requesting physician’s name, address, date of specimen collection, specimen source, and appropriate clinical information about the patient. *When the specimen arrives in the laboratory the laboratory staff affix an accession number or bar code label on each slide for further identification. *The laboratory should have written criteria for rejecting specimens. *Fixation while the specimen is still wet is recommended for conventional cell samples. *The Papanicolaou staining procedure is strongly suggested for most cytologic samples, unless additional staining procedures are warranted. *Staining solutions and chemicals used in the cytopathology laboratory should be labeled with the time of preparation, purchase, or both. Staining solutions should be filtered regularly to avoid contamination and should be covered when not in use. *Effective measures to prevent cross-contamination between gynecologic and nongynecologic specimens during the staining process must be used. 8. Slide Evaluation Workload:- *Regulations as to the number of specimens a cytotechnologist may evaluate in a 24-hour period are currently set at 100 slides per an 8-hour day. * This regulation may not do justice to the various conditions that influence the quality of the slide evaluation performance. * The percentage of atypical cases evaluated versus the percentage of negative cases in varying populations as well as screening of nongynecologic specimens should be considered when workloads are established. *This regulation ensures that the number and type of cytologic samples evaluated do not, through fatigue, adversely affect the cytotechnologist’s performance. 9. Cytologic Terminology:- *The vaginal/ectocervical/endocervical cytology sample should be interpreted preferably by using the Bethesda System. *The nongynecologic material should be interpreted in medical terms. 10. Laboratory Records, Logs, and Files:- *Each specimen should be recorded and a sequential accession number assigned together with the name of the patient and the originator of the sample. *Test records must be retained for at least 5-10 years. *The negative gynecologic cell samples should be retained on file for a minimum of 5 years and negative fine needle aspirates for 10 years or indefinitely if they exhibit abnormal features. *The modern cytopathology laboratory should use a computerized file system.

INTRODUCTION :- The use of fine-needle aspiration (FNA), a method of aspiration biopsy cytology, continues to grow throughout the World. Improvements in imaging, computed tomography scan (CT), and ultrasound (USG) have fueled the growth of FNAamong both radiologists and clinicians. The dominant clinical sites for FNA still remain breast, thyroid, and lymph nodes among superficial tissues. After reading this lesson, you will be able to: *describe the techniques of fine needle aspiration cytology                                                                                                              *arrange the clinic for performing FNAC                                                                                                                                                *assist the pathologist in performing FNAC                                                                                                                                          *smears and collect any fluids obtained from FNAC and process appropriately.  CLINICAL SKILLS REQUIRED :- Aspiration biopsy may be indicated whenever there is a palpable tumor mass ora lesion visualized within any organ. For the physician or more specifically for the pathologist performing FNA, some familiarity with general anatomy isessential. For the physician or more specifically for the pathologist performing FNA, some familiarity with general anatomy is essential. For the pathologist performing this biopsy some sharpening of clinical skills, both obtaining a  focused clinical history and performing a physical examination are required. Clinicians performing aspiration biopsy obviously lack this essential ingredient of experience and knowledge of morphology. Despite the recognized participation and value of cytotechnologists to an aspiration biopsy service, the pathologist must be actively involved in the aspiration biopsy, making both the initial and final evaluation of the smears. The Thin-needle Aspiration Method :- Thin needle generally 22, 23, 25, and 27 gauge, are used for the performance of aspiration biopsy, most often 1.5 in. in length. Special situations may dictate shorter needles and even higher gauge. For example, the very small cutaneousmetastasis of breast carcinoma on the chest wall may be sampled more easily with a 27-gauge, 1-in. or even ½-in. needle and with a small, 3.0- to 5.0-mL syringe, approaching the nodule in a plane perpendicular to the skin surface, inthe manner of performing a tuberculin skin test. Radiologists most often use the Chiba needle of 21 and 22 gauge for transthoracic and transabdominal aspirations. If one employs only the thin-needle technique, there are virtually nocomplications, the exceptions being FNA of the thorax (pneumothorax) or some cases of excessive bleeding with transabdominal aspiration biopsy. Basic Equipment :- The basic equipment used for rapid and efficient performance of thin-needle aspiration biopsy are as follows. 1. Cameco Syringe Pistol, Aspir-Gun, or other type aspiration handle; 2. 10 or 20-mL disposable plastic syringe with LuerLok or straight tip, depending on aspiration gun handle size; 3. 22 to 27-gauge, 0.6- to 1.0-mm external diameter disposable needles, 3.8 and 8.8 cm, 15 and 20 cm long, with or without stylus; the needle hub should be clear;  4. Alcohol skin preparation sponges; betadine skin sponges for deeper aspirations, transabdominal, transthoracic, bone (where the cortex is not intact or the periosteum is elevated), or deep soft tissue; 5. Sterile gauze pads 6. Microscopic glass slides with frosted ends; 7. Small vial of balanced salt solution and/or RPMI tissue culture transport media; 8. Suitable alcohol spray fixatives for immediate fixation of wet smears 9. 10 or 20 mL capped tube with 10% neutral buffered formalin for cell-block. 10. Optional vial of local anesthesia, 1-2% lidocaine; topical spray anesthesia for aspirates in children or intraoral aspirates; vials of lidocaine that dentists use for local anesthesia and the dispensing equipment may be useful; 11. Small vial of buffered glutaraldehyde for fixing aspirate for electron microscopy if required or anticipated. A small plastic tray easily holds all the equipment. Majority of the smears are to be air-dried and later stained with a Romanowsky method, the Diff-Quik stain being preferred. Some smears are usually wet-fixed in 95% ethyl alcohol. Aspiration Technique :- To be successful with an aspiration biopsy, it is important to follow the preliminary steps listed here: 1. Review the history of the patient. Determine the clinical problem and its relevance to the lesion to be biopsied. 2. Determine whether the biopsy is justified.  3. Palpate the mass, attempting to determine its location in relation to surrounding structures. Estimate its depth. Decide on the optimal direction of the needle to accomplish the aspiration biopsy. A mass located deeply in tissue in usually best approached perpendicularly to the skin surface. Small and superficially lying tumors are best approached by penetrating theskin at or very close to a horizontal plane, then feeling for the mass with the needle tip. 4. The patient should be placed in a comfortable position for the aspiration biopsy, but the mass must be easily palpable and immobilized during the biopsy. Step 4 is very important for head and neck lesions. The prominence of an enlarged lymph node, or lump, may sometimes depend on whether the patient is supine or erect. The sternocleidomastoid muscle bulk and its close proximity to the cervical lymph nodes require positioning the patient such that the biopsy needle passes through only a minimum of soft tissue and muscle before reaching