Principle of Medical L Aboratory Science Practice 2: Pmls 2 - PDFCOFFEE.COM (2025)

DOÑA REMEDIOS TRINIDAD ROMUALDEZ MEDICAL FOUNDATION COLLEGE OF MEDICAL TECHNOLOGY S.Y. 2021-2022

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PRINCIPLE OF MEDICAL LABORATORY SCIENCE 2 RTRMF- College of Medical Technology 2ND SEMESTER

Understanding Phlebotomy What is Phlebotomy? ● Greek Word: phleb = “vein” & tomia = “to cut/cutting” ●“To cut into a vein” ● Involves invasive procedures usually by cutting or puncture. ● Performed to obtain blood specimen for diagnostic testing either by: ➢ venipuncture (venous blood collection) ➢ arterial puncture (arterial blood collection) ➢ capillary/dermal puncture (capillary blood collection ● Blood collection for blood transfusion (blood banking) ● Used in therapeutic phlebotomy for some conditions.

Historical perspective ● Late Stone Age ➢ crude tools are used to cut blood vessels and drain blood from the body. ● 3500 B.C. ➢ Mesoamerican practice bloodletting for rituals and religious purposes rather than medicinal. ● 1400 B.C. ➢ EGYPT: Papyrus records and tomb paintings depicts bloodletting procedures using leeches. ➢ 1st to use bloodletting for medical purposes. ● Hippocrates of Cos (460-377. B.C.) & His

Contemporaries: GREECE ➢“Balance of 4 humours” ➢ Believed that an imbalance of blood, phlegm, yellow and black bile would cause disease. ➢ Treatment includes bloodletting, purging, catharsis, diuresis. ● Aelius Galenus/galen of pergamum

(129-200 A.D.): ROME ➢ Believed that blood was the dominant humour and the one in most need of control. ➢ Advocated the bleeding of the patients with leeches. ➢ Leeching or leechcraft persisted in standard treatment.

process of leeching 1. Place a drop of milk or blood on patient’s skin. 2. Introduce hirudo medicinalis (a European medicinal leech) to the site. 3. Allow the leech to fall off.

● Medieval Europe ➢ Bloodletting twas the best possible treatment during this time ➢ Monks and clergy men performed bloodletting to treat various conditions, from plague and small pox to epilepsy and gout until it was banned in 1163. ➢ Barbers began offering range of services that included: ◦ bloodletting ◦ cupping ◦ tooth extractions

◦ lancing

◦ amputations— along with, of course, trims and shaves thus calling them barbersurgeons. ➢ Pole - represented the rod squeezed by the patient to promote bleeding. ➢ White stripes - corresponded to the bandages, which were also used as tourniquets. ➢ 1800’s: Phlebotomy procedures were performed at its peak. A loss of approximately 10 mL (2 teaspoon) was standard.

Methods of Bloodletting ● Divided into: ➢ Generalized Method done by venesection and arteriotomy ➢ Localized Method done by scarification with cupping and leeches ◦ scarification- scraping the skin with a cube shaped brass box. ◦ cupping- heated suction apparatus creates a vacuum causing blood to flow into the cup. ● Venesection ➢ a process by which the median cubital vein is pierced with a sharp object (fleams or lancet) to drain blood. ◦ fleams- devices with multiple, variably sized blades that folded into a case like a pocketknife. ◦ thumb lancet/ lancet- small, sharp pointed, two-edged, instruments often with an ivory or tortoise shell case. ● Leeching or Hirudo Therapy ➢ use of medicinal leech (Hirudo Medicinalis), which can ingest about 5 to 10 mL of blood. Cañete, Shrine B.

● 19th century to present ➢ Leech therapy resurfaced and is used for microsurgery. ➢ Phlebotomy profession emerged as an expansion to Medical Technology ➢ Obtaining blood specimen collection for laboratory analysis and blood donor collection. ➢ Therapeutic profession.

Qualifications ● Philippines ➢ Ra 5527 Section 21 (Certification for Med Lab Technician) 1. Has finished a 2 year college course and has at least 1 year experience as medical laboratory technician. Provided that for every year of deficiency in college attainment, two years of experience may be substituted. 2. Has failed to pass the board examination for Medical Technology but obtained a general rating of 70%. ➢ Others: 1. Graduate of Medical Technology or Medical Laboratory Science 2. Graduate of other health profession and has undergone intensive training for phlebotomy (including blood bank phlebotomy) RRoles in the present healthcare society In the Philippines Health Care Setting they are: ➢ Tasked to collect blood specimens from patients/ clients laboratory analysis. ➢ If phlebotomist are assigned in the blood bank section, they are assigned in the screening and collections from donors. ➢ Carrying out therapeutic phlebotomy procedures. ➢ Preparation, accessioning, transport and handling of non- blood specimen. ➢ Performance of basic laboratory procedures, administrative and managerial task.

Traditional duties and responsibilities ➢ Demonstrate professionalism by their conduct, appearance, composure and communication skills. ➢ Know facility’s policies and procedures. ➢ Properly identify patients. ➢ Collect both venous and capillary blood specimen. ➢ Select the appropriate and accurate specimen container for specified tests. ➢ Properly label, handle and transport specimens following its policies. ➢ Sort specimens received and process specimen for delivery to each laboratory sections.

➢ Perform computer operations and/or update log sheets where require. ➢ Perform basic laboratory procedures including preparation of reagents, care and maintenance of glasswares and preparation of laboratory specimen for testing. ➢ Observe all safety regulations and always practice confidentiality. ➢ Attendance in a CPEs.

Additional roles of phlebotomist ➢ Training of other HCP to perform phlebotomy ➢ Monitoring the quality of samples collected on the units ➢ Evaluation of protocols associated with specimen collection ➢ Performing and monitoring of POCT ➢ Performing ECGs ➢ Performing of measuring patient’s vital sign ➢ Collection of arterial blood sample ➢ Collection of samples from central venous access devise

“Phlebotomists are Laboratory Ambassadors” Accuracy & Precision

Dependable, Cooperative & Committed

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Self Motivated

Good Communication Skills

PHLEBOTOMIST

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Compassionate, Courteous & Respectful

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Professional Apppearance

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Organized, Responsible & Flexible

Integrity, Honesty & Competence

Professional Traits Communication in healthcare setting ❖ COMMUNICATION BASICS ● Communication is the process by which information is exchanged or transmitted.

components: ● Verbal Skills ➢ introduction to patients, explaining the procedures, reassure the patient and assure the patient that procedure is accurately done. ➢ BARRIERS: ◦ hearing impairment ◦ patient emotions ◦ age & education levels ◦ non-english speaking ◦PWDs Cañete, Shrine B.

● Listening Skills ➢ Looking at the patient directly ➢ Encouraging the patient to express feelings, anxieties and concerns ➢ Allowing the patient to describe his/ her concerns ➢ Providing prompt feedback to the patient ➢ Encouraging patient communication by asking ● Non-verbal communication: Positive Gestures ◦ Body language ◦ Good body posture ◦ Good body posture ◦ Eye contact

Negative Gestures ◦ Dropping posture with head held below ◦ Looking down or away from the patient

◦ Neat, well-groomed

◦ Dingy, wrinkled lab coat;

appearance ◦ Respecting personal

too much jewelry ◦ Immediately approaching

space ◦ Cultural Diversity

patient’s space before greeting and explaining

Zone of Comfort 2 feet 2-4 feet 4-12 feet greater than 12 feet

Message

PHLEBOTOMIST

Communication Cycle

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Feedback

➢ Different Kinds of Patient ◦The excessively quiet ◦ The hostile ◦ The confused or disoriented ◦ The full of question ◦ The demanding ◦ The over friendly and seductive ◦ The opinionated

◦ The grieving ◦ The frustrated ➢ GOAL - Ensure that the patient was able to fully comprehend the process of the procedure to be performed this also includes assurance of patient comfort and other important instructions. ➢ ELEMENTS OF COMMUNICATION IN

HEALTHCARE: 1. Give recognition 2. Share information 3. Listen, do not just hear them 4. Demonstrate empathy, understanding and patience 5. Perform with comfort 6. Express gratitude

Philippine Healthcare facilities

● SECTION 7: Classification of Hospitals and other Health Facilities 7.1: Government or Private 7.2: General or Special 7.3: Service Capabilities

❖ COMMUNICATION & COSTUMER SERVICE

poor cation i commsuknills

◦ The scared and in pain

● Administrative Order No 147 S 2004: Revised Rules and Regulation Governing The Registration, Licensure and Operation of Hospitals and Other Health Facilities in the Philippines

procedure

➢ Intimate: ➢ Personal: ➢ Social: ➢ Public:

◦ The confident

PATIENT

anxiety

➢ Primary Care 1. Non-departmentalized hospital that provide clinical care and management on the prevalent diseases in the locality. 2. Clinical service includes general medicine, pediatrics, obstetrics and gynecology, surgery and anesthesia. 3. Provides appropriate administrative and ancillary services (radiology, laboratory pharmacy) 4. Provides nursing care for patients who require immediate, moderate and partial category of supervised care for 24 hours or longer. ➢ Secondary care 1. Departmentalized hospital that provide clinical care and management on the prevalent diseases in the locality, as well as particular forms of treatment, surgical procedure and intensive care 2. All clinical services in primary care as well as specialty care. Cañete, Shrine B.

3. All administrative and ancillary services in

THE CLINICAL LABORATORY

secondary care

4. Nursing care provided in primary care as well as total and intensive skilled care. ➢ TERTIARY CARE 1. Teaching and training hospital that provide clinical care and management provided in secondary care hospitals 2. All clinical services in secondary care as well as sub-specialty care. 3. All administrative and ancillary services in secondary care 4. Nursing care provided in secondary care as well as continuous and highly specialized critical care ➢ Infirmary a health facility that provides emergency treatment and care to sick and injured, as well as clinical care and management to mothers and newborn babies.

LABORATORY TESTS

➢ Birthing Home a health facility that provides maternity service on pre-natal and post natal care, normal spontaneous delivery and care of newborn babies. ➢ Acute-Chronic Psychiatric Clinic Facility a health facility that provides medical service, nursing service, pharmacological treatment and psychosocial intervention for mentally ill patients. ➢ Custodial Psychiatric Care Facility a health facility that provides long-term care, including basic human care services such as food and shelter to chronic mentally ill patients.

HOSPITAL ORGANIZATION

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Chemistry COagulation Hematology Microbiology Immunohematology/ blood bank Serology/ immunology

Cañete, Shrine B.

PRINCIPLE OF MEDICAL LABORATORY SCIENCE 2 RTRMF- College of Medical Technology 2ND SEMESTER

Infection Control, L aboratory Safety, First Aid and Personal Wellne! Infection control Infection is a condition that results when a microorganism (microbe for short) is able to invade the body, multiply, and cause injury or disease.

Definition of Terms: ● Infection ➢ colonization by a pathogen and establishing residence inside the body. ● Pathogen ➢ a microbial element that causes disease ➢ microbes that are pathogenic (causing or productive of disease are called pathogens ● Infectious/Communicable disease ➢ development of pathological manifestations from infectious agents (pathogens) that can be transmitted from one person to another. ➢ Communicable - able to spread from person to person ➢ Centers for Disease Control and Prevention - (CDC) an agency of the US Department of Health and Human Services - charged with the investigation and control of various diseases, especially those that are communicable and have epidemic potential. - develops guidelines and recommends safety precautions to protect healthcare workers and others from infections ● Nosocomial/Healthcare Associated Infection

➢ infection caused by a personnel, patients, visitors, food, drug, or equipment while a patient is in the hospital. ➢ Nosocomial Infection - applies to infections acquired in the hospital. ➢ Health Care Associated Infections - applies to infections associated with healthcare delivery in any healthcare setting, including home care. ➢ can result from contact with: ▸ infected personnel ▸ other patients ▸ contaminated food ▸ drugs ▸ visitors ▸ equipment ➢ UTI is the most common nosocomial infection IN US according to CDC. Accounting for 32% of all HAI

CHAIN OF INFECTION Infection transmission requires the presence of a number of components, which make up what is referred to as the chain of infection. 1. CHAIN OF INFECTION ➢ aka “causative agent” because this causes the development of pathological agents. ➢ pathogenic agent responsible for causing diseases: ▸ bacteria ▸ fungi ▸ protozoa ▸ viruses 2. reservoir ➢ source of an infectious agent ➢ “A place where microbes can survive, grow and, multiply” ➢ Reservoir Host - an individual or animal infected with a pathogenic microbe ▸ patients ▸ personnel ▸ visitors ▸ with active diseases ▸ incubating a disease ▸ chronic carrier ➢ Fomites - (inanimate objects) are major source of infectious agents depending on the following factors: a. amount of contamination b. viability of microbe c. virulence of microbe d. time of contamination Cañete, Shrine B.

3. Portal of exit ➢ exit pathway for the infectious agent to leave the host ➢ infectious agent can exit a reservoir host in secretions from: ▸ eyes ▸ nose ▸ mouth ▸ exudates from wounds ▸ tissue specimens ▸ blood from venipuncture ▸ skin puncture sites ▸ excretions of feces and urine 4. Means of transmission ➢ is the method an infectious agent uses to travel from a reservoir to a susceptible individual. ➢ means of infection transmission: ▸ airborne ▸ contact ▸ droplet ▸ vector ▸ vehicle ● Airborne Transmission

➢ dispersal of IA that are typically less than 5 μm and can remain infective for a longer period of time (INHALATION) ➢ generated by sneezing, coughing, talking and activities that generate aerosols ➢ patients with airborne infections require AIIRs; ➟ Airborne Infection Isolation Rooms ➟ should wear n95 and respirators ➢ examples are: ▸ mycobacterium tuberculosis ▸ rubeola virus ▸ varicella

➢ examples are: ▸ salmonella - infection from handling contaminated chicken ▸ shigella - infection from drinking contaminated water ▸ blood transfusion 5. Portal of Entry ➢ “the site of entry to the susceptible host” ➢ entry points: ▸ body orifices ▸ mucous ▸ membranes ▸ breaks in the skin ➢ invasive procedures: ▸ catheterization ▸ venipuncture ▸ capillary puncture 6. Susceptible Host ➢ someone who has decreased ability to fight infection ➟ Age (newborns and elderly) ➟ Health Status (vaccinated vs unvaccinated) ➟ Immune Status (hospitalized and/or immunocompromised)

Infection Control Programs “A typical infection control program implements procedure aimed at (1) breaking the chain of infection, (2) monitors and collects data of all infection occurring in the institution, and (3) institute special precautions in the event of outbreaks of specific infections” Elements of an Effective Infection Control Programs

● Droplet Transmission ➢ transfer of IA to mucous membranes of the susceptible host via infectious particles (> 5μm or larger) and cannot be suspended in air ➢ infectious droplets travels less than 10 mm ● Contact Transmission ➢ is the most common means of transmitting infections ➢ two types: ➟ Direct Contact - physical transfer (touching or kissing or sexual activity) ➟ Indirect Contact - contact with fomites (inanimate objects) ● Vector-Borne ➢ is the transfer of infectious agent carried by an insect arthropod or animal ➢ examples are: ▸ mosquitos ▸ fleas ● Vehicle Transmission ➢ transmission of IA through contaminated food, water, or drugs

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Screening and Protection of HCWs Infection Control Methods Standard Precautions Identification of Hazard and Minimizing the Risk 5. Evaluation, Incident Monitoring and Treatment of HCWs 6. Surveillance 7. Education and Training of HCWs 8. Research and Development 1.

Screening and Protection of HCWs ➢ screening of illnesses of the HCWs prior employment ➢ examples: ▸ tuberculosis (purified protein derivative and x-ray examination) ▸ hepatitis B infection (HBsAg and HBsAb) ▸ syphilis infection (rapid plasma reagin or anti-TP) ▸ salmonella or other diarrheal infection and skin diseases Cañete, Shrine B.

➢ immunization for different illnesses: ▸ HBV, measles, mumps, rubella (MMR), diphtheria, pertussis and tetanus (DPT) ▸ Vaccination can be done on or before employment. If within employment it is usually covered by the employer 2. Infection Control Methods ➢ Hand Hygiene most important means of preventing the spread of infection ROUTINE HAND WASHING Soap (antimicrobial soap), water Wash hands when visibly soiled Mechanical movement (friction) for 20 seconds

Chronology: 1. Palm to Palm 2. Right Palm over left dorsum with finger interlaced 3. Palm to Palm with finger interlaced 4. Inside of the Fingers with finger interlocked 5. Thumb 6. Fingers and Nails 7. Wrist

ALCOHOL DISINFECTANT Alcohol-based disinfectant (gels, foams and rinses), benzalkonium chloride, iodine solution Use disinfectant when hands are not visibly soiled Mechanical movement (friction) for 20 seconds

➢ Steps in Hand Wahing MUST-TO-REMEMBER!!!!!! If hand-washing is not possible, use a detergent containing wipes followed by alcohol-based disinfectant. 2. Perform handwashing after contact of sporeforming bacteria (Clostridium difficile and Bacillus spp.) 3. Surgical hand washing procedure must be performed for 2 mins with the use of antimicrobial soap (ICU, Burn Unit) 4. Natural nails should preferably less than 1⁄4 inch long. 5. Artificial fingernails or extenders must not be worn in ICU and other high risk areas. 1.

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➢ Hand Sanitation and Hand Washing

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MUST-TO-REMEMBER!!!!!! Gowns are disposable while coats can be reused!!! A properly worn gown has the sleeves pull all the way to the wrist, the belt tied, overlapped, closed and securely fastened. 3. Gown is removed by sliding the arms out of the sleeves slowly!!!

➢ Personal Protective Equipment (ppe) - Protective clothing and other items worn by individual to protect mucous membranes, airways, skin and clothing from contact with an infectious agent.

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a. GLOVES ➟ clean non-sterile or examination gloves ➟ latex type (common) ➟ vinyl/nitrile (allergic to latex) powdered/nonpowdered ➟ 3 main reasons why do we need to wear gloves: 1. Prevent contamination of hands when handling specimen or patient. 2. Prevent transmission of pathogen to the patient. 3. Prevent the possibility of transmission of IA from one patient to another. 1. 2.

MUST-TO-REMEMBER!!!!! Gloves should be pulled over the cuffs of the gown for adequate protection Glove removal: glove-in-glove and beaking method GOAL: NO GLOVE TO SKIN CONTACT

c. RESPIRATOR AND FACE MASKS ➟ MASK - is a face protection equipment worn to protect the mucous membrane (nose & mouth) against droplets generated by coughing and sneezing ➟ RESPIRATORS – NIOSH-approved N95 respirator that are required when entering patients with airborne precaution (e.g. px with TB) 1. 2.

MUST-TO-REMEMBER!!!!! WEARING THE MASK: (1) Ear loop must place properly, (2) adjust the metal/nose bar to fit snugly, (3) extend the mask to the chin WEARING OF THE RESPIRATOR: (1) Fasten the top ties around the upper portion of your head, (2) tie the lower one at the back of your neck, (3) press the nose bars to fit snugly – check for air leaks

d. FACE SHIELD AND GOOGLES ➟ Googles - is a face protection equipment worn to protect the eyes against droplets ➟ Face shield - cover the mucous membrane of the eye, nose and mouth from splashes or sprays of body fluid 1. 2.

MUST-TO-REMEMBER!!!!! Googles must cover the eyes properly for an absolute protection Face Shield must cover the entire face for a total protection!

➢ Donning of Personal Protective Equipment

b. LABORATORY GOWNS AND COATS ➟ Clean, non-sterile, fluid resistant cotton or synthetic materials ➟ Most gowns are adequately sized(below-theknee length)with knit cuffs and are backfastening ➟ Sterile gowns - are used in high-risk areas (ICU, NICU, Burn Unit) ➟ Laboratory coats - are required attire for most phlebotomy procedures.

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➢ Doffing of Personal Protective Equipent

➢ Nursery and Neonatal ICU Infection Control Technique ➟ Perform the Hand Hygiene and Donning of PPE in the ANTEROOM. ➟ Typical nursery and neonatal ICU infectioncontrol technique includes the following: ▸ Wash hands and put on clean gloves, gown, and mask. ▸ Gather only those items necessary to perform the specimen collection. ▸ Leave the blood collection tray in the anteroom outside the nursery. ▸ Remove gloves, decontaminate hands, and put on new gloves between each patient Same infection control techniques must be performed in High Risk areas such as Intensive Care Units, Burn Unit, Operating Rooms and Delivery Rooms. 3. ISOLATION PROCEDURES ➢ Minimizing the spread of infection by separating the patients with certain transmissible infections from contact with other patients and limit their contact with hospital personnel and visitors. CATEGORIES OF ISOLATION PROCEDURES

GOLDEN RULE: “Remove the most contaminated first then proceed to less contaminated equipment”

a. REVERSE /PROTECTIVE ISOLATION ➢ Reserved for patients with highly susceptible to infection (immunocompromised px) b. TRADITIONAL ISOLATION SYSTEM (diagnosis or suspicion must be established) ➢ Category-specific system – includes different 7 isolation categories often resulting to over-isolation of patients and needless extra costs ➢ Disease-specific system - was based on the modes of transmission of common diseases. c. UNIVERSAL PRECAUTION ➢ Introduced 1985 at the dawn of HIV Infection in HCWs ➢ “Blood and certain body fluids of all individuals were considered potentially infectious” d. BODY SUBSTANCE ISOLATION ➢ Integrates the elements of category specific and disease-specific isolation ➢ requiring that gloves be worn when contacting any moist body substance 4. GUIDELINES FOR ISOLATION PRECAUTION (two-tier method) ● Standard Precaution ➢ specifies precautions to use in caring for all patients regardless of diagnosis or presumed infection status ➢ Precaution is applicable to blood, all body fluids Cañete, Shrine B.

(including all secretions and excretions except sweat, whether or not they contain visible blood), non-intact skin, and mucous membranes. ● Transmission-Based Precaution ➢ used for patients known or suspected to be infected or colonized with highly transmissible or epidemiologically significant pathogens that require special precautions ➢ specifies precautions to use for patients either suspected or known to be infected with certain pathogens transmitted by airborne, droplet, or contact routes a. AIRBORNE PRECAUTION Applies when patients known or suspected to be infected with microorganisms transmitted by airborne droplet nuclei (particles smaller than 5 m) b. DROPLET PRECAUTION Applies when patients known or suspected to be infected with microorganisms transmitted by droplets (particles larger than 5 m) c. CONTACT PRECAUTION Applies when a patient is known or suspected to be that can be transmitted by direct contact with the patient or indirect contact with surfaces or patient-care items.

MUST-TO-REMEMBER!!!! The updated isolation precaution guideline added respiratory hygiene/cough etiquette as a new standard precaution. This precaution applies to all who enter a healthcare setting and includes covering the mouth and nose with tissue when coughing, prompt disposal of used tissues, hand hygiene after contact with respiratory secretions, and 3 feet of separation from persons with respiratory infections.

5. Evaluation and Treatment of HCWs ➢ This includes OSHA-mandated confidential medical evaluation, treatment, counseling, and follow-up as a result of exposure to blood-borne pathogens. 6. Surveillance ➢ This involves monitoring patients and employees at risk of acquiring infections as well as collecting and evaluating data on infections contracted by patients and employees. ➢ Infection-control measures are updated and new policies instituted based on this information. 7. Continuing Professional Education and Research ➢ Attendance in safety trainings offered within or outside the health institution ➢ Application of the modern trends for safety and infection control programs

Biosafety and biohazards ● BIOSAFETY ➢ The set of containment principles, technologies and practices that are implemented to prevent the unintentional exposure to hazards. ➢ “safe handling of biological substances to protect the HCW” ➢ is a term used to describe the safe handling of biological substances that pose a risk to h ● BIOHAZARD ➢ A biological material, situation, or act with a potential harm to health. ➢ Anything harmful are potentially harmful to health is called a biohazard (short for biological hazard) and should be identified by a biohazard symbol. ● SHARP HAZARDS ➢ Sources: ▸ needles ▸ lancets ▸ broken capillary tube ▸ glassware ➢ Precaution: (a) NEVER recap a needle! Use needle safety devices (holders, etc..) (b) All sharps must be disposed in a PUNCTURERESISTANT, leak- proof containers containing biohazard symbol. Cañete, Shrine B.

Biosafety ● BIOHAZARD EXPOSURE ROUTES ➢ Airborne: - inhalation of aerosols, splashes (droplets), aerosols and fumes ➢ Ingestion: - neglect in hand hygiene, “chewing of pencils or pen”, “licking fingers when turning pages of the book” ➢ Non intact Skin: - visible and pre-existing breaks of the skin (cover with water-proof bandage) ➢ Percutaneous: - needlestick injury (most common to phlebotomists) ➢ Permucosal: - infectious agents entry through mucous membrane (eyes, mouth, nose). ● BLOOD BORNE PATHOGENS (BBP) ➢ Any infectious microorganism present in patient’s blood and other body fluids even if not having signs and symptoms of the disease ➢ SIGNIFICANT CONCERNS: ▸ HBV ▸ HCV ▸ HIV ➢ Other BBPs: ▸ other hepatitis viruses ▸ Cytomegalovirus (CMV) ▸ syphilis ▸ malaria ▸ prions ▸ West Nile Virus (most recent) ● Exposure control plans ➢ Promulgated by OSHA Blood Borne Pathogen Standard & Needlestick Safety & Prevention Act of 2001 ➢ The standard requires implementation of engineering controls and practice work controls

● EXPOSURE INCIDENT PROCEDURE ➢ Needlestick and other sharps injury: - Carefully remove the needle or glass shards and wash the site with soap and water for a minimum of 30 mins. ➢ Mucous Membrane Exposure: - Flush the site with water or sterile saline for a minimum of 10 minutes. - Remove contact lenses and disinfect theme if being reused or discarded. ➢ Report immediately to the supervisor and to a medical professional for medical evaluation, counseling and treatment. ● POST EXPOSURE PROPHYLAXIS

● SURFACE DECONTAMINATION ➢ OSHA requires ALL surfaces for specimen collection and processing areas to be decontaminated. ➢ 1:10 dilution of bleach (NaOCl) solution (or any recommended by OSHA & EPA) ➢ Cleaning must be done before and after every shift wearing gloves. ● CLEAN UP BODY SPILLS ➢ Special EPA-approved chemical solutions and kits available for cleaning up of blood and body fluid on surfaces. ● BIOHAZARD WASTE DISPOSAL ➢ Blood and other body fluids must be disposed in biohazard labeled container ➢ Nonreusable items that are contaminated with blood must be disposed.

Electrical safety

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WHAT TO DO IF ELECTRIC SHOCK OCCURS: 1. Turn off circuit breaker and unplug the equipment 2. If it cannot be shut off, move the equipment using a non conductive glass or wood. 3. Start CPR to the patient if needed. 4. Keep the victim warm

● Class K ➢ fires occur with high-temperature cooking oils, grease, or fats and require agents that prevent splashing and cool the fire as well as smother it.

FIRE SAFETY Fire components ● Fuel - combustible material ● Heat - to raise the temperature of the material until it ignites or catches fire ● Oxygen - to maintain combustion or burning have traditionally been referred to as the fire triangle ● Chemical Reaction - produces the fire, actually creates the fire tetrahedron

Fire extinguisher ➢ There is a fire extinguisher class that corresponds to each class of fire except class D. ➢ Class D fires present unique problems and are best left to firefighting personnel to extinguish. ➢ Some fire extinguishers are multipurpose to eliminate the confusion of having several different types of extinguishers. ➢ Multipurpose extinguishers are the type most frequently used in health-care institutions. classes:

● Class A extinguishers ➢ use soda and acid or water to cool the fire. ● Class B extinguishers ➢ use foam, dry chemical, or carbon dioxide to smother the fire. Classes of Fire - five classes of fire are now recognized by the National Fire Protection Association (NFPA)

● Class C extinguishers ➢ use dry chemical, carbon dioxide, Halon, or other nonconducting agents to smother the fire.

● Class A ➢ fires occur with ordinary combustible materials, such as wood, papers, or clothing, and require water or water-based solutions to cool or quench the fire to extinguish it.

● Class ABC (multipurpose) extinguishers ➢ use dry chemical reagents to smother the fire. They can be used on class A, B, and C fires.

● Class B ➢ fires occur with flammable liquids and vapors, such as paint, oil, grease, or gasoline, and require blocking the source of oxygen or smothering the fuel to extinguish

● Class K extinguishers ➢ use a potassium-based alkaline liquid specifically formulated to fight high-temperature grease, oil, or fat fires by cooling and smothering them without splashing. Some class K extinguishers can also be used on class A, B, and C fires.

● Class C ➢ fires occur with electrical equipment and require nonconducting agents to extinguish. ● Class D ➢ fires occur with combustible or reactive metals, such as sodium, potassium, magnesium, and lithium, and require dry powder agents or sand to extinguish (they are the most difficult fires to control and frequently lead to explosions). Cañete, Shrine B.

● HAZARD COMMUNICATION (HazCom) ➢ To protect employees who may be exposed to hazardous chemicals ➢ According to the law, all chemicals must be evaluated for health hazards and all chemicals found to be hazardous must be labeled as such and the information communicated to the employees ➢ The HazCom standard is known as “The Right to Know Law” because of the labeling 4 Essential Requirements: 1. Statement of Warning 2. Statement of Hazard 3. Precautions 4. First Aid Measures ● Material safety data sheet (MSDS) ➢ Contains the general information as well as precautionary and emergency information of the product. ● Department of Transportation Labeling System

Radiation safety ● The principles involved in radiation exposure are distance, shielding, and time. ● A phlebotomist may encounter radiation hazards ➢ when collecting specimens from patients who have been injected with radioactive dyes, ➢ when collecting specimens from patients in the radiology department or nuclear medicine, and ➢ when delivering specimens to radioimmunoassay sections of the laboratory. ● This symbol must be displayed on the doors of all areas where radioactive material is present

Chemical safety ● A phlebotomist may come in contact with hazardous chemicals when using cleaning reagents, adding preservatives to 24-hour urine containers, or delivering specimens to the laboratory.

Cañete, Shrine B.

● National Fire Protection Association Hazard

Degree of Hazards 0 = No/Minimal Hazard 1 = Slight Hazard 2 = Moderate Hazard 3 = Serious Hazard 4 = Extreme Hazard

FIRST AID ● EXTERNAL HEMORRHAGE 1. Apply Pressure with Hands EXPOSE to find where the bleeding is coming from and apply FIRM, STEADY PRESSURE to the bleeding site with both hands if possible. 2. Apply Dressing and Press EXPOSE to find where the bleeding is coming from and apply FIRM, STEADY PRESSURE to the bleeding site with bandages or clothing. ● Shock ➢ a state of shock results when there is insufficient return of blood flow to the heart, resulting in an adequate supply of oxygen to all organs and tissues of the body. Common Symptoms: ▸ Pale cold, clammy skin ▸ Rapid, weak pulse ▸ Increased, shallow breathing rate ▸ Expressionless face and staring eyes First Aid for Shock: 1. Maintain an open airway for the victim. 2. Call for assistance. 3. Keep the victim lying down with the head lower than the rest of the body. 4. Attempt to control bleeding or other cause of shock if known. 5. Keep the victim warm until help arrives. ● CARDIOPULMONARY RESUSCITATION AND EMERGENCY CARDIOVASCULAR CARE ➢ 6-8 hours of Basic Life Support Training: CPR Training on all ages and Automated external defibrillator (AED)

➢ Certification is good for 2 years THINGS TO REMEMBER: ▸ A compression rate of at least 100 per minute ▸ An adult compression depth of at least 5 cm (2 inches) ▸ An infant and child compression depth of one third of the anterior-posterior diameter of the chest; approximately - 4 cm (1.5 inches) for infants - 5 cm (2 inches) for children ▸ Allowing complete chest recoil between compressions ▸ Minimizing interruptions in chest compressions (trying to limit them to less than 10 seconds) ▸ Avoiding excessive ventilation (BLS single rescuer compression-to-ventilation ratio stays at 30:2) D = danger R = response A = airway B = breathing D = defibrillation

S = shout for help C = circulation

Personal wellness ● Personal hygiene - Essential - Healthcare workers work in close contact with others so body odor must be controlled - Daily shower, deodorant, oral hygiene, clean undergarments - Strong odors caused by tobacco, perfumes, scented products, etc. can be offensive ➢ Nutrition ➟ has been defined as the “act or process of nourishing.” In other words, a food is nutritious if it supplies the nutrients the body needs “to promote growth and repair and maintain vital processes.” ➢ Rest and Exercise ➟ Healthcare workers often complain of fatigue (physical or mental exhaustion). ➟ Fatigue brought on by physical causes is typically relieved by sleep. ➟ Being physically fit increases the chance of staying healthy and living longer. 3 Components of measuring Fitness: ✓ Strength - the ability to carry, lift, push, or pull a heavy load ✓ Flexibility - the ability to bend, stretch, and twist ✓ Endurance - the ability to maintain effort for an extended period of time Cañete, Shrine B.

➟ Exercise strengthens the immune system, increases energy, and reduces stress by releasing substances called endorphins, which create a peaceful state.

Key terms ● CSLI ➢ Clinical and Laboratory Standards Institute ● OSHA ➢ Occupational Safety and Health Administration ● BBP ➢ Blood-borne Pathogen ● HAI/LAI ➢ Healthcare Associated Infection ➢ Laboratory Acquired Infection ● CDC ➢ Center for Disease Control and Prevention ● NIOSH ➢ National Institute of Occupational Safety and Health ● HazCom ➢ Hazards Communication ● MSDS ➢ Material Safety Data Sheet ● PPE ➢ Personal Protective Equipment ● HIV HCV HBV ➢ Human Immunodeficiency Virus (HIV) ➢ Hepatitis C Virus (HCV) ➢ Hepatitis B Virus (HBV)

Cañete, Shrine B.

PRINCIPLE OF MEDICAL LABORATORY SCIENCE 2 RTRMF- College of Medical Technology 2ND SEMESTER

Blood Collection Equipment, Additives, and Order of Draw ● Phlebotomy ➢ “an incision into a vein” ➢ the collection of blood samples for laboratory analysis to diagnose an monitor medical condition. ➢ in preparing a patient for phlebotomy, there should be care to minimize physiologic factors that affect laboratory determinations: ➟ diurnal variation - concentration of some blood constituents is affected by the time of day ex: CORTISOL (peaks at 8AM) ➟ fasting - most common factor that affects the determination of blood - FBS (fasting blood sugar) - lipid profile ➟ exercise ➟ diet ➟ ethanol ➟ tobacco consumption ● Venipuncture ➢ the most common way to collect blood specimens that can be performed by: ➟ evacuated tube system (ETS) ➟ syringe ➟ winged infusion set (butterfly)

Evacuated tube system ➢ most frequently used method for venipuncture ➢ a collection system that uses vacuum to pull blood into a container process:

➟ the blood is directly collected to the evacuated tube ➟ eliminating the need for transfer of specimen that minimizes the risk of biohazard exposure ➢ this consist of a multisample needle, a tube holder and color coded evacuated tubes.

➟ Double Pointed Needle - to puncture the stopper of the collection tube ➟ Tip - is where you insert into the vein

➟ Holder - to hold the needle and the blood collection tube (has a color coded evacuated tubes). ➟ Rubber Sleeve - or the rubber sheath of the puncturing end of the needle is screwed securely into the small opening of the holder. ➟ Flange - the opposite end of the holder; this is where you insert the evacuated tube ● MULTISAMPLE NEEDLES ➢ the ETS needles are designed as multi sample needles because they have the stopper puncturing needle that is covered by a rubber sleeve or a rubber sheath → is pushed back when a needle is attached and it returns to it’s full needle coverage when the tube is removed → this characteristic prevents the leakage of blood when tubes are being changed → when tubes are removed it is removed with a slight twist to help disengage them from the needles → the manufacturers of this needles have a blunting device to provide additional protection against needlestick injuries by making the needles blunt before removing from the patient ➢ allows collection of multiple tubes during venipuncture ➢ threaded - so it can screw into a tube holder ➢ beveled point on each end ➢ the tube of the needle is covered by a sleeve that retracts allowing blood flow and recovers the needle to prevent leakage when tube is removed. ● Tube Holder ➢ are made of rigid plastic and may be designed to act as a safety shield for used needles ➢ plastic cylinder with a small opening for needle at one end

Cañete, Shrine B.

and a large opening for tubes at the other. ➢ these holders are available to accommodate the different sizes of collection tubes ➢ the tubes has flanges to help place and remove tubes. ● Evacuated Tubes ➢ tubes used for collection are called the evacuated tubes or vacutainers → have pre-measured vacuum that automatically draws the volume of blood indicated on the label

l

→ it is an evacuated tube holder with a rubber sheathed needle inside.

➢ if the blood to be tested requires an anticoagulant: → speed is very important for the phlebotomist to be able to transfer the blood from the syringe before clot formation begins. → once the blood has been transferred, the anticoagulated tube must be thoroughly mixed to avoid small clot formation.

➢ these tubes is fully inserted to the end of the holder when the needle is in the vein, blood will flow into the tube once the needle penetrates this stopper

➢ Tube Stoppers → this is the stopper end of the tube and you insert the tube into the rubber sheeted needle and the blood now flows from the vein directly to the tubes → stated on the tube is the draw volume, vacuum in the tube, and the expiration date so the manufacturers guarantee us the integrity of the anticoagulants/additives → are color coded to identify the type of anticoagulant, additive, absence of additive or special tube property → the color coding indicates the type of sample that will be obtained when a particular tube is used.

Syringe system ➢ Often preferred over ETS for patients with small or fragile veins. ➢ The advantage of this system is that phlebotomists will be able to control the suction pressure on the vein by slowly withdrawing the syringe plunger. ➢ Blood drawn in a syringe is immediately transferred to appropriate evacuated tubes to prevent the formation of clots. ➢ It is not advisable to insert the syringe needle into tube stopper of the vacutainer. unless provided by a blood transfer device → provides a safe means for blood transfer without using the syringe needle or removing the tue stopper.

➢ syringes routinely used for venipuncture range from 2-20 mL, and a size corresponding to the amount of blood needed to be used. ➢ consist of a; → barrel graduated in milliliters (mL) or cubic centimeters (cc) → plunger that fits tightly within the barrel, creating a vacuum when retracted. → needles used with syringes are attached to a plastic hub designed to fit onto the barrel of the syringe ➢ are individually packaged, sterile, and color coded as to gauge size.

Butterfly system ➢ also known as “winged infusion sets” ➢ used when performing venipuncture from very small veins that is usually seen in pediatric or geriatric patients ➢ to accommodate the dual purpose of venipuncture and infusion, the needle is attached to a flexible plastic tubing that can then be attached to an IV setup, syringe, or specially designed evacuated tube holders. ➢ are usually 21, 23, or 25 gauge with lengths of 1/2 to 3/4 inch → short needle with a plastic part resembling butterfly wings → plastic wings attached to the end of the needle to aid the insertion of the needle into the small veins → tubing is attached to the back end of the needle which terminates with an adapter for the attachment of a syringe or an evacuated tube holder → has a length of tubing with Luer fitting (syringe use) or a Luer adapter for ETS use Cañete, Shrine B.

➢ this plastic attachments resemble the butterfly wings used for holding the needle during insertion or to maneuver the needle when working with very small veins ➢ during use, the plastic wings are typically held together with the thumb and index finger, allowing the user to achieve the shallow needle angle needed to access small veins

Materials Needed for venipuncture ● Gloves ➢ are worn to protect healthcare workers from contamination by the patient’s body substances and to protect the patient from possible microorganisms brought by the healthcare workers hands ➢ Wearing gloves is not a substitute for hand washing ➢ variety of gloves is available including; ▸ sterile and non sterile ▸ powdered and unpowdered ▸ latex and non latex (vinyl, nitrile, neoprene, and polyethylene) ● Skin disinfectants/ antiseptics ➢ ALCOHOL → Isopropyl alcohol and ethyl alcohol - are commonly used in routine venipuncture as well as capillary puncture → Alcohol pads - are much preferred because they are individually packed and can be utilized or opened only when he dead ➢ POVIDONE IODINE → strong disinfectant used as skin disinfectant for bacterial culture as well as in blood donation phlebotomy → the most frequently used solutions are povidone-iodine and tincture of iodine (for persons who are allergic to iodine) ➢ BENZALKONIUM CHLORIDE → preferred disinfectant for alcohol level tests because alcohol should not be used when dealing with ethanol examinations ● Tourniquet ➢ fastened to a patient arm to restrict venous flow but not arterial flow ➢ this distends the veins making them visible and palpable ➢ stretches the walls of the veins so they are thinner and easier to pierce ➢ the most frequently used tourniquets are: ▸ flat latex ▸ vinyl strips → they are inexpensive and may be disposed or reused

→ flat nonlatex strips are available for patients or phlebotomists allergic to latex ➢ should only be applied for 1 minute and not longer because there are some laboratory examinations that would be affected with a long application ➢ fastened 3 - 4 inches above the venipuncture site ➢ if the tourniquet is closer to the site, the vein may collapse as blood is above the venipuncture site ➢ blood pressure cuffs can also be used as tourniquets → used primarily for veins that are difficult to locate → inflated to a pressure of 40 mmHg and this allows the blood to flow into but not out of the affected veins ● Needles ➢ needle gauge refers to the diameter of the needle bore ➢ the gauge of the needle is related to the size of the needle; → the larger the gauge, the smaller the needle bore and length → large (16-gauge) needles — used to collect units of blood for transfusion → small (23-gauge) needles— used for very small veins ➢ if the needle is: → too large = needle can damage the vein → too small = needle may cause hemolysis to the blood ➢ 21 gauge needle is standard for venipuncture ➢ 23 gauge needle is used for children and for small and difficult veins ➢ Needle length: 1 - 1.5 inches ➢ Butterfly needle: 1/2 - 3/4 inch ➢ Manufacturers package needles individually in sterile twist-part sealed shields that are color coded by gauge for easy identification. → needles should be visually examined before use to determine for structural defects like: ▸ non beveled points ▸ bent shafts → needles must not be used if seal is broken or defective ▸ needles should not be recapped once the shield is removed regardless if it was used or not. → needle structure varies to adapt to the type of collection equipment being used. → all needles consist of: ▸ bevel (angled point) ▸ shaft ▸ lumen ▸ hub Cañete, Shrine B.

● Tube additives ➢ most ETS tubes contain additives or anticoagulant that prohibits clot formation ➢ tests that require whole blood or plasma are collected in tubes containing anticoagulants to prevent clotting of the sample → if the additive is an anticoagulant, the blood will not clot and the specimen will be whole blood that can be centrifuged to obtain plasma → anticoagulant - prevent clotting by binding calcium or inhibiting thrombin in the coagulation cascade ➢ most all other additives and additive free tubes (e.g. glass red top) produce serum specimens ANTICOAGULANTS ➟ OXALATE ➢ combines with calcium to form an insoluble salt ➢ concentration: 1-2 mg/mL of blood ➟ CITRATE ➢ combines with calcium in a non-ionized form ➢ concentration: 3.2-3.8 g/dL in a ratio of 1 part to 9 parts of blood or 1:4 ➢ inverted: 3-4x ➢ coagulation test require the correct ratio of blood and anticoagulant ➢ containers or the coagulation tubes need to be filled adequately → an insufficient blood volume leaves an excess citrate in the plasma and results to falsely elevated clotting time. ➢ sodium citrate is the required anticoagulant for coagulation studies as it preserves the labile coagulation factors → ratio of blood to the liquid sodium citrate is critical and should be 9 to 1 → over mixing a light blue stopper tubes can activate platelets and cause’ erroneous coagulation test results ➟ ETHYLENEDIAMINE TETRAACETIC ACID (EDTA) ➢ chelates calcium

➢ preparation: Versene and Sequestrene ➢ in K2EDTA OR K3EDTA ➢ concentration: 1-2 mg/mL of blood ➢ inverted: 8x ➢ EDTA is the anticoagulant of choice for hematology cell counts and cell morphology that is our CBC or the complete blood count ➢ available as a spray dried dipotassium or liquid tripotassium salt ➢ Coagulation is prevented by binding of calcium in the sample to the sites of the large EDTA molecule thereby preventing the participation of the calcium in the coagulation cascade. ➢ for hematology procedure that require whole blood, such as CBC, dipotassium EDTA is the anticoagulant of choice because: → maintains the cellular integrity better than other anticoagulants → inhibits platelet clumping → does not interfere with routine staining procedures ➢ used specifically for blood bank in some facilities ➟ HEPARIN ➢ preparation:▸ lithium ▸ sodium ▸ potassium ➢ concentration: 0.2 mg/L of blood ➢ an effective anticoagulant in small quantities without significant effects on many determination ➢ act as an antithrombin & antithromboplastin ➢ they are the ideal universal anticoagulant and is available in lithium heparin and sodium heparin → heparin - accelerates the action of antithrombin - it neutralizes thrombin and prevent the formation of fibrin ➢ primarily used for chemistry test performed on whole blood or plasma particularly the STAT test → short turnaround time test that requires a fast a results ANTIGLYCOLYTIC AGENTS ➟ FLUORIDE ➢ are available with a variety of anticoagulants and additives for the primary purpose of preserving glucose ➢ usually used for glucose measurements because they contain a preservative or an antiglycolytic agent (sodium fluoride) → sodium fluoride prevents glycolysis for three days → 2 mg NaF/mL of blood prevents glycolytic enzyme for up to 48-71 hours → not an anticoagulant ➢ anticoagulant: potassium oxalate → 10 mg of NaF/mL could act as an anticoagulant by forming weakly dissociated Ca components Cañete, Shrine B.

CLOT ACTIVATORS ➢ does not contain additives resulting for the blood to clot ➢ used for most chemistry blood bank and immunology assays ➢ red stopper plastic tubes contain silica as a clot activator ➢ coagulation factors such as: ▸ thrombin and: ▸ substances like glass (silica) particles &; ▸ inert slays like diatomite (Celite) that enhance clotting by providing more surface for platelet activation ➢ blood that is collected in tops or red glass tubes clots by normal coagulation process in about 60 minutes and centrifugation of the sample then yields serum as the liquid portion ➢ red stopper tubes are used for the same purpose as the red plastic tubes and there is no need to invert glass red stopper tubes THIXOTROPIC GEL SEPARATORS ➢ serum separator tubes that contains thixotropic gel at the base of the tube where blood is forced into during centrifugation → inert substance contained in or near the bottom of certain tubes ➢ this gel undergoes a temporary change in viscosity during centrifugation and lodges the pact cells from the serum → the gel forms a barrier between liquid phase (serum) and the cells because of its density of 1.04 g/cm3 → physically separates the liquid component of the blood from the cells (1.03 and 1.09 g/cm3 respectively) after centrifugation SPECIAL TUBES COLOR Yellow Yellow

ADDITIVE SPS Acid Citrate Dextrose

Blood Culture HLA Phenotyping, Paternal Testing Trace elements, toxicology, Nutritional Studies

None

White

EDTA and gel

Molecular Diagnostic

Black

Sodium Citrate (4:1)

ESR (Westergren)

K2EDTA

ABO & Rh typing, Ab screening (Blood Bank)

(With crossmatch label)

➢ black stopper tubes → contains buffered sodium citrate they are used for Westergren sedimentation rates → they differ from the light blue tops in that they provide a ratio of blood to liquid anticoagulant of 4 to 1 → specially designed tubes for Westergren sedimentation rates are available

Order of draw ORDER OF DRAW

TUBE STOPPER COLOR

RATIONALE FOR COLLECTION ORDER

Blood culture (sterile collections)

Yellow SPS Sterile media bottle

Minimizes chance of microbial contamination

Coagulation tubes

Light blue

The first additive tube in the order because all the other additives affect coagulation tests

Red Glass non additive tubes

Prevents contamination by additives in other tubes

Plastic clot activator tubes Serum separator tubes (SSTs)

Red

Plasma separator tubes (PSTs)

Green and gray rubber Light green plastic Green

Heparin affects coagulation test and interferes in collection of serum specimens; causes the least interference in tests other than coagulation tests

EDTA tubes

Lavender Pink Pearl top

Responsible for more carry over problems than any other additive: elevates Na and K levels, chelates and decreases calcium and Iron levels, elevates PT and PTT results, sodium fluoride and potassium oxalate affect sodium and potassium levels, respectively after hematology tubes because oxalate damages cell membrane and causes abnormal RBC morphology. Oxalate interferes in enzyme reaction

CLINICAL USE

Royal Blue

Pink

→ used to collect samples to be cultured for presence of microorganisms → prevents coagulation by binding calcium

Plasma preparation tubes (PPTs) Oxalate/ fluoride tubes

Red and gray rubber Gold plastic

Gray

Filled after coagulation tests because silica particles activate clotting and affect coagulation tests (carry over of silica into subsequent tubes can be overridden by anticoagulant in them

➢ yellow stopper tubes → anticoagulant: sodium polyanethol sulfonate Cañete, Shrine B.

➢ length: 1.75 mm ➢ depth of the incision: → infants and children: < 2.0 mm → adults: < 2.5 mm ~~ to avoid contact with the bone ~~

Capillary puncture Dermal or skin puncture is especially useful for pediatrics where removal of larger quantities of blood can have serious consequences. ➢ the method of choice for most institutions in collecting blood from infants ➢ because pediatrics patients may have veins that cannot be punctured by small gauge needle and drawing excess amount of blood from premature or small infants can cause anemia ➢ certain test require capillary bloods such as: ▸ newborn screening test ▸ capillary blood gasses ➢ due to advancements in laboratory instrumentations and popularity of point of care testing made it possible to perform a majority of laboratory test on micro samples of blood obtained by dermal or capillary puncture Compositions of capillary blood ➢ blood collected by dermal puncture comes from the capillaries, arterioles, and venules → therefore, it is a mixture of arterial and venous blood and may contain small amounts of interstitial and intracellular fluids ➢ blood is obtained by puncturing the capillary bed of the skin with a lancet or other sharp device ➢ collection site: → adults/children: 3rd or 4th fingers → infants (newborn): heel of the foot ➢ because of arterial pressure, the composition of this blood more closely resembles arterial rather than venous blood ➢ warming the site before sample collection increases blood flow as much as sevenfold → thereby producing a sample that is very close to the composition of arterial blood Materials for capillary puncture ● Lancets ➢ sterile, disposable, sharp instruments used for capillary puncture

● Microtainer ➢ special small plastic tubes often referred to as “bullets” ➢ variety of anticoagulants and additives, including gel separator, are available. ➢ are color coded in the same way as evacuated tubes → colors correspond to color coding of evacuated tubes and marking for minimum and maximum fill levels typically measured in microliters (µL) ➢ some tubes are supplied with a capillary scoop collector top that is replaced by a plastic color coded sealer top after the sample is collected ➢ designed to hold approximately 600 µL of blood ➢ specimens must be collected quickly for capillary puncture: → to minimize the effects of platelet clumping and microclot formation → to ensure that an adequate amount of specimen is collected before the bleeding stops Order of Filling: Hematology specimens are collected first because they are most affected by clotting: 0. Slides 1. EDTA specimens 2. Other additive specimens 3. Serum specimens ● Microhematocrit Tubes ➢ capillary tubes, which are frequently referred to as microhematocrit tubes ➢ small tubes used to collect approximately 50 to 75 µL of blood for the primary purpose of performing microhematocrit test ➢ are designed to fit into a hematocrit centrifuge and its corresponding hematocrit reader ➢ available plain or coated with ammonium heparin and are color coded in: → heparinized (red) - for collecting directly from a capillary puncture → nonheparinized (blue) - to be used when filling with blood from an EDTA tube ● Slides ➢ thin glass that are used to place liquid specimens for microscopic viewing Cañete, Shrine B.

PRINCIPLE OF MEDICAL LABORATORY SCIENCE 2 RTRMF- College of Medical Technology 2ND SEMESTER

Review of !e Circulatory and Lymphatic System Circulatory System

➢ is an integral part of the human body. ➢ is responsible for transporting oxygenated and deoxygenated blood

Functions of Circulatory System 1. Transports oxygen and other nutrients to the tissues. 2. Removes waste metabolic product of the cells and the carbon dioxide gas for excretion. 3. Fighting diseases and infection caused by microbial pathogens, cancer cells or even by the autoimmune diseases. 4. Maintenance of the homeostasis by acid-base regulation. 5. Homeostasis and coagulation action. ➢ this is made up of two components: → cardiovascular system: ▸ heart ▸ blood vessels ▸ blood → lymphatic system: ▸ lymph ▸ lymph nodes ▸ vessels

Different parts of the circulatory system Heart ➢ muscular organ that is essential for life ➢ pumps blood through the body ➢ it is a hollow cone shaped organ about the size of a fist and measures an average of 14 cm long and 9 cm wide → a healthy adult, at rest, pumps approximately 5L of blood per minute ➢ has four chambers which is surrounded by a fluid filled sac called the pericardium → atrium: left & right - upper chamber which collects blood → ventricle: left and right - lower chamber which pump blood functions: ➟ right atrium receives deoxygenated blood from the body and pumps it to the right ventricle and into the pulmonary artery ➟ left atrium receives oxygenated blood from the

lungs and pumps it to the left ventricle which then pumps the blood into the aorta LAYERS OF THE HEART 1. Epicardium ➢ thin, watery membrane on the outer layer of the heart ➢ it covers the heart and is attached to the pericardium 2. Myocardium ➢ thick layer of cardiac muscles in the middle layer of the heart ➢ pumps blood into the arteries by contracting 3. Endocardium ➢ thin layer of epithelial cells in the inner layer of the heart ➢ lines the valves and interior chambers FUNCTIONS OF THE HEART ➢ Generating blood pressure - which is required to force blood through the blood vessels ➢ Routing blood - to ensure flow of oxygen rich blood to tissues ➢ Ensuring one way blood flow - through the heart and blood vessels ➢ Regulating blood supply ~~ heart receives blood supply through the left and right coronary arteries ~~ coronary veins return the oxygen between the blood from the heart muscle back to the heart

Blood vessels ➢ Blood that flow through the blood vessels is regulated so that: → cells receive adequate nutrients → waste products are removed FUNCTIONS OF THE BLOOD VESSELS ➢ carries blood ➢ exchange nutrients, waste products and gases with tissues ➢ transport substances ➢ helps regulate blood pressure ➢ directs blood flow to the tissues Cañete, Shrine B.

TYPES OF BLOOD VESSELS 1. Venous System ➢ act as a capacitance vessels or blood reservoirs of the body ➢ thin walled tubes that carry deoxygenated blood from the tissues towards the heart → dark red colored blood ➢ veins are said to “merge”, “converge”, “merge” and grow larger as they go near the heart ➢ venules are very small veins that collect blood from the capillaries ➢ 3 superficial veins in the arm → cephalic vein - 2nd site that you can consider - though the size is significant it is however a movable vein which can be a problem during veni → median cubital vein - preferred site for venipuncture - the most prominent and stable vein among the 3 → basilic vein - least preferred site - aside from being a movable vein, it is near the brachial artery which can be punctured during the insertion 2. Arterial System ➢ or arteries are thick walled blood vessels that carry oxygen rich blood from the heart to the tissues → bright red colored blood ➢ arteries are said to “diverse”, “branch”, and “fork” as they form smaller divisions away from the heart ➢ arterioles are thinner blood vessels that transport blood from the arteries t the capillaries 3. Capillaries ➢ fine hair like blood vessels that connect arterioles and veins ➢ smallest of the vessels and is also the thinnest because it’s composed of overlapping endolethium producing gaps of slits ➢ this is where the metabolic exchange between the blood and tissue takes place → blood flows from arteriols into the capillaries which branch out to form a network, as blood flows to the capillaries, blood gives up oxygen and nutrients to the tissue and take up carbon dioxide and other metabolic products of metabolism

ARTERIES VS VEINS Arteries

Veins

Carry blood from the heart, carry oxygenated blood (except pulmonary artery

carry blood to the heart, carry deoxygenated blood (except pulmonary vein

Normally bright red in color

Normally dark red in color

Elastic walls that expand with surge of blood

Thin walls/less elastic

Can feel a pulse

No pulse

Different parts of the circulatory system ➢ blood → is the fluid that is transported through out the body via the circulatory system → composed of plasma and other formed elements

BLOOD COMPONENTS 1. FATTY LAYER ➢ mostly composed of lipoproteins such as chylomicrons or sometimes very low density (VDL) 2. PLASMA ➢ is composed of: → 90% water → 8% plasma proteins ▸albumin ▸globulin ▸fibrinogen → 2% solutes ➢ clear, straw colored liquid portion of the blood → plasma - anticoagulated; inhibit clot → serum - from coagulated tube ➢ triglycerides can sometimes cause turbidity in the plasma fluid which is mostly affected by fasting

3. FORMED ELEMENTS ➢ buffy coat → upper yellowish layer: PLATELETS → lower yellowish layer: WBCs ➢ packed erythrocytes Cañete, Shrine B.

● LEUKOCYTES ➢ also known as white blood cells ➢ colorless nucleated cells that circulate in the peripheral blood which primary function is to defend the body from foreign invaders ➢ present in much smaller number than erythrocytes in peripheral blood ➢ according to granularity, they are classified as: → granulocytes - contain distinct granules in their cytoplasm → nongranulocytes - lack the prominent granules ~~granules that are present are the primary identifying characteristic of leukocytes~~ ➢ stained blood smear → the major routine method for studying the morphology of leukocytes with the use of Romanowksy Stain such as: ➟ Wright’s stain - which the granules are named according to their staining reaction ➢ RR: 4.5 - 11.0 x 10 / L 5 Types of Leukocytes: ➢ Neutrophil → polymorphonuclear leukocyte or also called the “PMN” → also the most common leukocytes → nucleus: segmented into 5 lobes → cytoplasm: stains light pink → the neutrophil granules remain neutral → showing minimal affinity for either acidic or basic dye and appears light pinkish or purple → purpose: to protect the body against infection → phagocytosis (main mechanism) - the process of locating, ingesting, and killing bacteria and other foreign invaders → RR: 40 - 60% in a normal peripheral blood ➢ Lymphocytes → play a major role in the maintenance of health and in response to and recovery of diseases → nucleus: large, round, and dark staining mostly encompassing most of the cell → cytoplasm: barely visible and ranges in staining intensity from deeply basophilic to pale blue → some lymphocyte migrate to the thymus where they become t cell which are the immunocompetent in cellular immunity → other precursor of lymphocyte develop into b cells which fulfill the function of

humoral immunity or antibody production → RR: 20 - 40 % in the peripheral blood ➢ Monocyte → also called “mononuclear phagocyte” - since phagocytosis is one of its main functions → nucleus: highly variable, round (horseshoe shaped) its distinct characteristic is kidney bean shaped → cytoplasm: abundantly staining which colors dull, pale or faded gray blue that contains fine indistinct granules → RR: 1 - 6% in peripheral blood ➢ Eosinophil → nucleus: a nucleus similar to neutrophils → cytoplasm: filled with granules staining bright orange pink because they have an affinity for eosin, (which is the acidic dye in the stain, usually evenly distributed but rarely overly the nucleus → functions as phagocytes but appear to move slower and have less intercellular killing ability than neutrophil → eosinophil plays a major role in parasitic infection and hypersensitivity reaction → eosinophilia - an increase in eosinophil could project allergic reaction → RR: 0 - 6% in the peripheral blood ➢ Basophil → least common of the leukocyte → makes up 0 - 1% in the peripheral blood → phagocytic capacity of basophil is substantially less than that of neutrophil and eosinophil → important function is the role of immediate hypersensitivity reactions → nucleus: light purple staining, that may be round invented band shaped or longulated usually difficult to see because of the overlying granules → cytoplasm: characterized densely by dark violet or purple dark granules that have an affinity for methylene blue (the basic dye in the stain) → very distinct because of its purple to almost black granules ➢ pH of the staining system or the Wright’s Stain → which is used for staining peripheral blood is very important for proper cell identification. Cañete, Shrine B.

● PLATELETS ➢ also known as “thrombocytes” → are cytoplasmic fragments of megakaryocyte with an average size of 2-4 µL ➢ plays a vital role in homeostasis ➢ primary action is to make a clot through platelet plug formation mechanism by releasing granules ➢ with Wright’s Stain → they have a light violet purple granular appearance that looks like specs of dots ➢ RR: 150 - 400 x 10 9 /L ● ERYTHROCYTES ➢ also known as red blood cells ➢ small biconcave disk-shaped anucleated cells with an average size of 7.2 µL ➢ with Wright’s Stain → it has a central pale area also called “central pallor” that is seen which fades gradually into reddish pink cytoplasm → this central pallor corresponds to the indentation in the erythrocyte disk ➢ filled with hemoglobin that carries oxygen and nutrients to the tissue and removes waste and carbon dioxide from the tissue ➢ RR: 3.8 - 6.5 x12/L

➢ if this excess fluid does not go back it can cause edema which would result to tissue damage and eventually death 2. Fat Absorption ➢ the lymphatic system absorbs fat and other substances from the digestive tract through the lacteals 3. Defense ➢ microorganisms are filtered from lymph by the lymph nodes and from blood by the spleen

Lymph ➢ is clear, watery fluid that is collected from the intercellular spaces ➢ transported through the lymphatic vessel, ducts and masses of lymph tissue called nodes ➢ often contain fats (chyle) when it comes from intestinal organs ➢ is filtered by passing through several lymph nodes before entering the venous system ➢ passes through the lymph nodes, which produces lymphocytes, before it reaches the ducts

Lymphatic nodes ➢ are organized collections of lymphatic tissue permeated by lymph channels ➢ produce lymphocytes and plasma cells and filter the lymph ➢ trap bacteria drained from an infected area and contain reticuloendothelial cells and phagocytic cells (macrophages) that ingest these bacteria ➢ are hard and often palpable when there is a metastasis and are enlarged and tender during infection.

Lymphatic vessels

Lymphatic system ➢ a network of tissues and organs responsible for the removal of toxins and waste in the body ➢ primary function is to transport the WBC to and from the lymph nodes throughput the body ➢ stores lymphocytes and monocytes to protect the body by phagocytosis and immune response

Functions of lymphatic System 1. Fluid Balance ➢ an excess interstitial fluid enters the lymph and passes through the lymphatic vessel to return to the blood

➢ serve as one-way drainage toward the heart and return lymph to the bloodstream through the thoracic duct or the right lymphatic duct ➢ carry lymphocytes from lymphatic tissues to the bloodstream. ➢ have valves which are constricted at the sites of valves showing a beaded appearance ➢ absorb large protein molecules and transport them to the blood stream because these molecules cannot pass through the walls of the blood capillaries ➢ they serve as one way #$$% towards the heart and return them to the bloodstream through the thoracic duct

Lymphatic capillaries ➢ begin blindly in most tissues, collect tissue fluid, and join to form large collecting vessels that pass to regional lymph nodes ➢ absorb lymph from tissue spaces and transport it back to the venous system ➢ they are called lacteals in the villi of the small Cañete, Shrine B.

intestine, where they absorb emulsified fat.

Laboratory tests and correlations

● Hematocrit ➢ for Anemia ➢ goes hand in hand with platelet count when a patient is having dengue fever ● Hemoglobin ➢ for Anemia ● Platelet Count ➢ for bleeding tendencies ● Prothrombin Time (PT) ➢ for coagulation disorders

● Troponin I and T ➢ most common for myocardial infarction ➢ is a STAT exam for doctors to know if patient is having a heart a attack

● APTT/ PTT ➢ activated partial thromboplastin time ➢ goes hand in hand with prothrombin time because they are the two most commonly performed test for coagulation disorders (Heparin Therapy)

● White Blood Cell (WBC) Count ➢ for infections or leukemia

● Bleeding Time (BT) ➢ also correlated with bleeding time and clotting time (Platelet Function) → performed when patient undergoes surgery ● Blood Culture ➢ for Microbial Infection in the blood ● CRP ➢ or C-reactive protein ➢ for Inflammatory Disorder which is also a marker for acute bacterial infection ➢ for covid 19 diagnostic testing this is one of the test for diagnosis if having bacterial infection ● CBC ➢ complete blood count ➢ most common and primarily for bleeding disorders, anemia, or leukemia ● Cholesterol ➢ for coronary artery disease ● ESR ➢ can be requested for inflammatory disorders when CRP is not available Cañete, Shrine B.

Principle of Medical L Aboratory Science Practice 2: Pmls 2 - PDFCOFFEE.COM (2025)
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