HISTOPATH SEM 1

• Diagnostic Cytology


  Microscopic examination of cells from different sites of the body for diagnostic purposes
• Divisions of Diagnostic Cytology

  • Exfoliative cytology
  • Fine Needle Aspiration
• Exfoliative cytology


  Microscopic study of cells that have been desquamated from epithelial surfaces
• Applications of Exfoliative cytology

  • Assessing malignant or cancerous conditions
  • Detection of asymptomatic cancer in women
  • Assessment of female hormonal activity in case of sterility and endocrine disorders
  • Determination of genetic sex (Barr bodies)
  • Determination of the presence of possible infection
• Fine Needle Aspiration (FNA)


  Study of cellular samples obtained from organs that do not shed cells spontaneously
• FNA


  • Simple, safe, and rapid cytologic technique that has been commonly used for the diagnosis of cancer
  • FNA of superficial masses is usually done by clinicians, or in some centers by the pathologist
• Specimens for examination


  • Vaginal smear
  • Endometrial and endocervical smears
  • Prostatic and breast secretions
  • Gastric or bronchial secretions
  • Pleural and peritoneal fluids
  • Sputum
  • Smears of urine sediment
  • Cerebrospinal fluid

  
• Sample Preparation for an optimal specimen


  Prepared smears should be fixed immediately, by quick immersion in a fixative or by a fixative spray
• Sample Preparation if smears cannot be made immediately

  1. Collected material should be placed in 50% alcohol

  2. This can be replaced with Saccomano's fixative (50% alcohol + 2% carbowax)

  
• Adhesive agents for cytologic method

  • Pooled Human Serum or Plasma
  • Celloidin Ether Alcohol
  • Leuconostoc Culture (group of microorganism)
  • APES (Aminopropyltriethoxysilane)
• Sample Preparation for specimens such as urine, serous effusion and watery lavage

  1. Optimum amount: 20-30mL

  2. Cells may be viable for up to 5 days (if refrigerated at 4 deg C)

  3. They require concentration (cytocentrifugation) of cells before transferring to glass slides

  4. Centrifuged ASAP at 1000 RPM speed for 1 minute and supernatant is removed

  5. A smear is made in an egg albumin-coated glass slide

  6. Fix the smear using 95% alcohol
• Smear Techniques in Exfoliative Cytology

  • Streaking
  • Spreading
  • Pull apart
  • Touch or impression smear

  
• Cytologic Smear Fixatives


  • 95% ethyl alcohol
  • Ether alcohol
  • Carnoy's Fixative
  • Commercial aerosol spray (e.g hairspray)
• Papaniculaou (Pap's) smear


  Staining method of choice for exfoliative cytology, developed by Dr. George Papanicolaou to identify stages in the maturation of exfoliated squamous epithelial cells and screen for cervical cancers
• Pap's Stain


  Polychromatic staining reaction with components: Harris Hematoxylin (nuclear stain), OG-6 Stain (Cytoplasm of mature cells), EA 50 (Cytoplasm of Immature cells)
• Advantages of Pap's Stain

  • Transparent blue staining of cytoplasm is obtained
  • Excellent nuclear detail is produced
  • Color range is predictable and of great value in the identification and classification of cells
  • Valuable in comparing cellular appearances in smears
• Modified Papanicolaou Staining Procedure

  Provides optimum nuclear detail information
• Non-Gynecologic Specimens


  • Peritoneal, pleural, pericardial fluids
  • Cerebrospinal fluid
  • Gastrointestinal specimens (gastric lavage, gastric brush, FNA for mucosal lesions)
  • Urine
  • Breast secretion
  • Respiratory Tract Specimens (Sputum, BAL/BW, BB)

  
• Bartlett's Classification


  Scheme for grading the quality of sputum specimens after a slide of the specimen had been examined at 100x magnification, assigning + and - values
• Vaginal Hormonal Cytology


  Relatively inexpensive, may be performed regularly even in pregnant women without undue risk, taken from the upper lateral third of the vaginal wall
• Ferning


  Technique used to determine the presence of estrogen in the uterine cervical mucus, to test for ovulation and early detection of pregnancy
• Risk management


  The process of ensuring and maintaining personal as well as environmental health and safety in the laboratory
• Risk management process


  Identify all electrical, mechanical and biological hazards that can potentially cause harm in the laboratory
• Types of hazards

  • Biological
  • Chemical
  • Fire/Explosive
  • Electrical
  • Sharps
  • Radioactive
  • Physical
  • Ergonomic
  • Cryogenic Material
• Biological hazard

  • Source: Infectious agent
  • Possible Injury: Bacterial, viral, parasitic, fungal
• Chain of infection

  1. Infectious agent

  2. Reservoir

  3. Portal of Exit

  4. Means of transmission

  5. Portal of Entry

  6. Susceptible host
• Hand hygiene


  Break the chain of infection
• Hand hygiene procedures

  • Wash with soap and water for 20 seconds if hand is visibly soiled
  • Use alcohol-based sanitizers if hand is not visibly soiled
• Standard precautions


  Combined universal precaution and body substance isolation procedures; Applies to all blood and body fluids and stresses handwashing
• Universal precaution


  All patients were assumed to be carriers of bloodborne pathogen
• Body substance isolation


  Required gloves to be worn when encountering blood or any body substance
• Chemical hazard


  • Source: Reagents, stain, etc.
  • Possible injury: exposure to toxic/carcinogenic agent
• Safety Data Sheet (SDS/MSDS)


  Major source of safety information for employees who may use hazardous materials in their occupation
• NFPA Diamond

  Degree of hazard: 4-Extreme, 3-Serious, 2-Moderate, 1-Slight, 0-No/Minimal
• Operating a fire extinguisher


  PASS: Pull the pin, Aim the base of the fire, Squeeze the handle, Sweep nozzle side to side
• Classes of fire

  • Not provided

  
• Other hazards include electrical, sharps, radioactive, physical, ergonomic, and cryogenic material