prelim lab

• Icteric:

  • Color: Dark to very dark yellow-brownish
  • High bilirubin and ALT/SGPT
  • Alcoholic baby
• Lipemic:

  • Color: Turbid to milky consistency
  • High triglyceride and carbohydrate
  • Chyle - a milky bodily fluid consisting of lymph and emulsified fats, or free fatty acids
• Hemolyze:

  • Color: Red sera
  • Red destruction of red blood cells
  • Unacceptable: Automatic repeat collection
  • Causes: Wrong collection, probing, too much tube inversion
• Pipettes:

  • Used for the quantitative transfer of reagents and the preparations of serial dilutions of specimens such as serum
  • Manual pipetting is prone to human error
  • Hand pipetting using fixed-volume is fast in small applications and only requires the hand of a practiced lab tech instead of extra hardware
  • Time-consuming, results can be unreliable, repetitive actions can lead to injury
  • 5-10 samples per hour
  • Always be mindful of the meniscus
• Proper Pipetting Techniques:

  • Adjust pipette to the correct volume
  • Check if you are using the correct pipette tip
  • Make sure no bubbles are produced
  • Always use the pipette in a vertical position
  • No reusing of pipette tips
  • Common pipetting errors: Loose pipette tip, tilting the pipettor, plunger quick release causing air bubbles, second stop draw
• Immunity:

  • Immunity is the body's ability to prevent the invasion of pathogens or resist harmful microorganisms
• Types of Immunity:

  • Innate:
  • General barriers
  • Physical Barrier: skin, stomach acid, macrophage, neutrophil
  • Biological barrier
  • Adaptive
• Differences between Innate and Adaptive Immunity:

  • Innate Immunity:
  • Presence already present in the body
  • Specificity: Non-specific
  • Rapid
  • Potency: Limited and lower potency
  • Memory: No memory
  • Allergic Reaction: Immediate and delay hypersensitivity
  • Adaptive Immunity:
  • Created in response to exposure to a foreign substance
  • Specific
  • Slow (1-2 weeks)
  • Potency: High potency
  • Memory: Long term memory
  • Allergic Reaction: Allergic Reaction
• Natural Immunity vs. Adaptive Immunity:

  • Natural (Innate):
  • Non-specific
  • No Memory
  • Examples: Exogenous (Skin), Endogenous (Stomach Acid), Phagocytosis (PMNS), Natural Killer Cells (NK)
  • Adaptive (Acquired):
  • Specific
  • Memory
  • Examples: T Cells (cytokines), B Cells (antibodies)
• Immune System:

  • Innate
  • Adaptive
• Types of Immunity: Active vs. Passive

  • Active: Individual produces antibody
  • Passive: Antibody transferred to individual
• Types of Immunity:

  • Active: Individual produces antibody, follows immunization or infection
  • Passive: Antibody transferred to individual, examples include Gamma Globulin injections and placental transfer
• Active Immunity:

  • Stimulates the body's immune system
  • Involves antibodies or cell-mediated immunity, or both, to protect against infectious agents
  • Examples include vaccines or toxoids
• Passive Immunity:

  • Provides temporary protection through the administration of exogenously produced antibody
  • Examples include transplacental transfer and immunoglobulin injections for specific purposes
• Physical components of the immune system:

  • Skin
  • Cough reflex
  • Tears (enzyme in tears)
  • Mucosal layer
  • Stomach acid
• 1. Lymphocytes:

  • Helper T cells
  • Suppressor T cells
  • Cytotoxic T cells
• Phagocytes:

  • Monocytes
  • Macrophages
  • Neutrophils
  • NK cells
  • Dendritic cells
• 1. Lymphocytes:

  • Clonal B cells
  • Memory B cells
• Comparison of Types of Acquired Immunity:

  • Active Natural: Infection, antibody produced by host, long-lasting immune response
  • Active Artificial: Vaccination, antibody produced by host, long-lasting immune response
  • Passive Natural: Transfer in vivo or colostrum, no antibody produced by host, short immune response
  • Passive Artificial: Infusion of serum/plasma, no antibody produced by host, short immune response
• Natural Immunity:

  • Natural Passive: Antibodies from mother to fetus or through milk, short-lasting
  • Natural Active: Exposure to live pathogen, develops disease, becomes immune
• Artificial Immunity:

  • Artificial Passive: Injected antibodies from a different person or animal
  • Artificial Active: Antigen presented, antibodies fight and remain alert for future infection, vaccine
• Immunizing Agents:

  • Substances or organisms that provoke an immune response when introduced into the body
  • Types include vaccines, immunoglobulins, and antisera
• Characteristics of Effective Vaccines:

  • Safety
  • Protection at the population level
  • Long-lasting effects inducing T and B cell memory
  • Cost-effective
  • Easy administration with no side effects
• Antisera and Antitoxins:

  • Specific immunoglobulins prepared from plasma of immunized animals or humans
  • Examples include snake venom, anti-tetanus, and rabies vaccine
• Immunoglobulins:

  • IgG: Greatest concentration in serum, activates complement, crosses placenta
  • IgM: Largest antibody, fixes complement best, prominent in early immune response
  • IgA: Predominant in body secretions, primary defense against local infections at mucosal surface
  • IgD: Unknown function, present on B cell surface
  • IgE: Allergy, involved in release of histamines from mast cells
• Constituents of Immunizing Agents:

  • Suspending fluid: sterile water or saline
  • Preservatives, stabilizers, antibiotics: inhibit bacterial growth, stabilize the antigen
  • Allergic reactions may occur if the recipient is sensitive to any of these additives
  • Adjuvants: enhance immune response to vaccines containing inactivated microorganisms
• • Kits designed to work with various sample types like blood, serum, urine, or swabs for versatility in different settings
  • User-friendly Design:
  • Simple kit operation with clear instructions and minimal steps for users with varying expertise
• Considerations when using IS test kits:

  • Antigen-Antibody Specificity:
  • Testing kits rely on the specific interaction between antigens and antibodies for accurate and targeted detection of the analyte of interest
  • Sensitivity and Specificity:
  • Testing kit should detect the analyte at low concentrations (sensitivity) and accurately distinguish it from other substances (specificity)
  • Rapid and Efficient Detection:
  • Delivering quick and efficient results through visual indicators or automated processes for timely decision-making
  • Sample compatibility:
• Safety Procedures:

  • Laboratory Safety Guidelines:
  • Conduct yourself responsibly in the laboratory
  • Follow all written and verbal instructions carefully
  • Do not touch equipment or materials until instructed
  • Avoid eating, drinking, or chewing gum in the laboratory
  • Be prepared, read procedures thoroughly, and avoid fooling around
  • Universal Precaution:
  • Treat all human blood and certain body fluids as infectious
  • Responsible for Hospital Waste Management:
• • Hospital staff, government agencies (DOH, DENR, League of City, Municipal, Provincial Mayors, Association of Barangay Council), Non-Government Office (NGO) promoting environmental protection
• Pipetting Techniques:

  • Check specimen before using the pipette to avoid lipemia, hemolysis, bacterial contamination
  • Icteric or turbid serum may interfere with results
  • Collect blood specimens before meals to avoid chyle
  • Avoid contamination with alkali or acid
  • Colors to remember and expect:
  • Icteric sample: dark yellow, brownish; high bilirubin or ALT/SGPT
  • Lipemic sample: turbid, milky; high cholesterol and triglycerides
  • Hemolyzed sample: red, repeat collection if needed
• Manual Pipetting:

  • Hand pipetting with fixed-volume for small applications, but can be time-consuming and lead to injury
  • Always mindful of the meniscus for accurate readings
  • Mouth pipetting is hazardous and unacceptable in the clinical laboratory

  Semi-Automatic Pipettes:

  • Scale up production incrementally and increase reproducibility
  • Technician moves hand probe from vessel to vessel
  • Can handle 11-100 samples per hour

  Automation:

  • Valuable for high-throughput applications, processing hundreds of samples at a time without human intervention
  • Protects from hazardous/infectious samples
• Types of Pipettes:

  • Transfer pipettes: Volumetric pipette, Ostwald folin pipette
  • Graduated pipette: Serological pipette, Mohr pipette
  • Micropipette for rapid repetitive measurements and delivery of predetermined volumes
• Proper Pipetting Techniques:

  • Adjust pipette to correct volume
  • Use correct pipette tip
  • Avoid producing bubbles
  • Always use pipette in a vertical position without reusing tips

  Common Pipetting Errors:

  • Loose pipette tips
  • Tilting the pipettor
  • Plunger quick release causing air bubbles
  • Second stop draw