Clinical Internship 1

Internship 1 - EACMED Transcribed by: AAA

TOPIC 1: BIOSAFETY & BIOSECURITY patients (e.g. respiratory secretions,

Speaker: Sir Judah P. Celeste, RMT blood, or body fluids)
 4-6 hours = change face mask
FUNDAMENTAL CONCEPTS OF LAB BIOSAFETY
AND BIOSECURITY - Used during procedures requiring
1. BIOSAFETY sterile technique to protect patients
- containment principles, technologies, and practices, from exposure to infectious agent
implemented to prevent UNINTENTIONAL exposure/ - Worn on the face
accidental exposure to pathogens and toxins, or their  Covers at least the nose and
unintentional release. mouth
 Keep bad bugs from people  Reduce the risk of inhaling
hazardous airborne particles
- Safety measures that helps to avoid infecting oneself (including dust particles, and
and others (or the environment) when handling infectious infectious agents), gases, or
substances RESPIRATOR vapors

- Use of PPE to be safe from harmful incidents once you - Health worker should undergo fit
are exposed from biological hazards testing regularly

- You are protecting yourself from the pathogens - N95 mask = a commonly used DR
2. BIOSECURITY
- the protection, control, and accountability for valuable - Always perform a USER SEAL
biological materials including information in laboratories in CHECK
order to prevent unauthorized access, loss, theft, - Goggles or a disposable face shield
misuse, diversion, or INTENTIONAL release of that covers the front and side of the face
pathogens and toxins FACE
 Keep bad people from bugs SHIELD - Personal eyeglasses and contact
lenses are NOT considered adequate
- You are protecting the biological materials from bad eye protection
people - Change gloves
 Ex. bioterrorism  Torn
 Heavily contaminated
PRINCIPLES OF BIOSAFETY  In bet. patients and procedure
1 Containment
Recognize and Assess Biological Hazards and - Use clean gloves when handling pens
2 GLOVES and other didactic materials
Laboratory Associated Infections
3 Promote Safe Microbiological Practices
- Remove and discard gloves when
1. CONTAINMENT leaving the patient room or care area
- Preventing the release of the infectious agent to the
environment and it is achieved thru: - Always perform hand hygiene after
 Safety Equipment: design to protect from injury removing gloves
or the spread of infection or illness. ORDER OF DONNING AND DOFFING
 Facility Design: take into consideration the DONNING DOFFING
movement of people, specimens, and laboratory 1 Hand Gloves Shoe cover
waste and equipment 2 Inner gloves Outer gloves
 (Good) Laboratory Practices: most important 3 Coverall Face shield
element of laboratory containment 4 Respiratory Coverall
A. PPEs 5 Face shield Respiratory
- designed to protect from injury or the spread of infection 6 Shoe cover Inner gloves
or illnesses 7 Outer gloves Hand hygiene
 Lab gown SAFETY EQUIPMENTS FACILITY DESIGNS
 Face mask - Biosafety cabinets
 Respirator - PPE - Basic laboratory
 Face shield - Enclosed containers - Containment laboratory
 Gloves - Vaccination (provide - Maximum Containment
increased level of Laboratory
- Put on clean isolation gown upon
protection)
entry into the patient room or area
PRIMARY BARRIERS SECONDARY BARRIERS
- Change the gown if it becomes soiled Most Important Element/ Concept:
LAB GOWN GOOD MICROBIAL LABORATORY PRACTICES
- Properly discard used or
contaminated lab gown into a BIOLOSAFETY CABINET
dedicated container or contaminated - ventilated enclosure
linen before leaving the patient area  Class: I, II, III
FACE MASK - Protects healthcare workers from  Level: 1, 2, 3, 4
contact w/ infectious materials from - also known as

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 Clinical Internship 1
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 Fume hood - Levels of containment range from the lowest to highest

 BSC,  BSL 4 – Post life threatening
 Laminar flow hood / PCR cabinet  BSL 3 - Exotic agents which may cause serious/
potentially lethal disease
-effective primary containment  BSL 2 - Disease causing agents of moderate
potential hazard
-clearance of open space in front of BSC – 1 METER  BSL 1 - Not consistently known to cause disease
- SHOULD be located out of the lab personnel traffic
pattern

- do not modify BSC in a way that may compromise its

containment unless approved by appropriate safety
personnel

- documentation such usage logs, certification records are

vital

- use of open flame and high pressure gases is NOT

encouraged in a BSC
BSL DEFINITION
 Causes disruption in HEPA filter
defined and with well-characterized strains and
not known to cause disease in humans
- surface decontamination with bleach or its equivalent
surface disinfectant is most effective  Minimal potential hazard
HEPA FILTER (HIGH EFFICIENCY PARTICULATE AIR)
BSL-1 - Ex.
- CONTINUOUS stream of inward air that prevents
 E.coli
aerosols from escaping through the front opening
 Saccharomyces cerevisiae
- Capable of filtering greater than 99.7% of airborne  K-12
particles smaller or larger than 0.3 diameter  Non-infectious bacteria
Moderate potential hazard. Mild disease to
- TEST humans
 SMOKE PATTERN TEST – confirm inward flow
or to ensure correct flow of air -Ex
BSL-2  Hepatitis A virus,
 FILTER LEAK TEST – scan the filter at a phase
of 1-2 inches/second using an aerosol parameter  Strep. pyogenes,
 Borrelia burgdorferi (Lyme disease),
- MECHANISM  Salmonella spp
 Diffusion - small particles Indigenous or exotic. Potentially lethal disease
 Interception – mid range size particles through respiratory transmission
 Impaction – large heavy particles
BSL-3 - Ex.
 Yersinia pestis (plague),
 Mycobacterium tuberculosis,
 SARS (COVID-19)
Dangerous and exotic. Fatal and without
treatment of vaccines
BSL-4 -Ex.
 Ebola virus,
CLASS I, II, III BSC COMPARISON  smallpox virus
RELATION OF RISKS FROUPS TO BIOSAFETY
LEVELS, PRACTICES, AND EQUIPMENTS

BIOSAFETY LEVEL
- A level of the biocontainment precautions required to
isolate dangerous biological agents in an enclosed facility
CORE REQUIRMENT FOR BSL 2 LABORATORY
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1 Good Microbiological Practice and Procedure - Protection, control and accountability for valuable
2 Personnel Conference and Training biological materials within the laboratories to prevent
3 Specimen receipt and storage unauthorized access, loss, theft, misuse, diversion or
4 Facility design intentional release
5 Emergency Response plan
6 Biosafety Cabinet and Laboratory Equipment BIORISK ASSESSMENT
7 Decontamination and waste Management - Considered as the backbone of biological risk
8 Occupational Health management, biosafety, and biosecurity.
9 Personal Protective Equipment
10 Controlled/Limited access to laboratory - Performed as often as needed
11 Clean to dirty work flow
ENHANCEMENT FOR BSL 2 LABORATORY -CONSIST OF
Directional Airflow  Risk Assessment BEGINS with: HAZARD
1 - Airflow from low area of contamination to higher IDENTIFICATION
area of contamination o DONE by BIOSAFETY OFFICER
2 Training And Proficiency  Define Preventive Measures
3 Working In Buddy System  Sound Allocation of Resources
4 Strict Monitoring of Equipment and Facility  Basic Legal Requirement
5 Strict Monitoring of Body Temperature AMP MODEL
6 Restriction Of Work to Authorized Personnel Only Assessment
 risk identification
PROPER HANDWASHING 1  hazard threat identification
1 Wet hands with running water  likelihood evaluation
2 Apply enough soap to cover all hand surfaces  consequence evaluation
3 Rub hands, palm to palm Mitigation
Right palm over left dorsum with interlaced fingers,  elimination/ substitution
4  engineering controls
and vice versa
2
5 Palm to palm with fingers interlaced  administrative controls
Back of fingers to opposing palms with fingers  practices and procedures
6
interlocked  PPE
Rotational rubbing of left thumb clasped in right palm Performance
7
and vice versa  control
Rotational rubbing, backwards and forwards with 3
 assurance
8 clasped fingers of right hand in left palm, and vice  improvement
versa RISK MANAGEMENT PROCESS
9 Rinse hand with water 1 Identify the hazard
1 Dry hands thoroughly with single use towel 2 Assess the risk
0 3 Control the risk
1 Use of towel to turn off faucet 4 Review the control plan
1 HAZARD VS THREAT
HAZARD THREAT
STEPS IN WASTE MANAGEMENT - anything in the - PERSON W/ means
1 Segregation environment that causes motives and opportunities
2 Collection something bad or to cause and access, theft and
3 Treatment harm misuse of isolates and
4 Storage  Physical biological materials
5 Transport  Material  Intentional
6 Final Disposal  Activity
MITIGATION
BIOLOGICAL WASTE COLOR CODING - Actions and control measures that are put into place
COLOR TYPE OF to reduce or eliminate the risks associated with
TYPE OF WASTE
CODE CONTAINER biological agents and toxins
Non-infectious DRY - most effective
BLACK waste ELIMINATION
- physically remove the hazard
Trash bin/ plastic (boxes, papers) - replacement of the procedure
bag Non-infectious WET SUBSTITUTION or biological agent with a similar
GREEN waste entity
(food) ENGINEERING - physical changes in work
Infectious/ CONTROLS stations, equipment’s,
Plastic bag
YELLOW Pathological Waste production facilities, etc.
(double bagging)
(tubes, cottons)  includes facility design,
Sharp isolate workers from
RED Puncture-proof
(syringes, needles) the hazard
ORANGE Containers Radioactive
- AIR HANDLING = the air must
BIOSECURITY flow from an area of lower

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 Clinical Internship 1
Internship 1 - EACMED Transcribed by: AAA

contamination to an area of
higher contamination
 inward directional
airflow
- change the way work is
performed
 Approved SOP
(Standard Operating
ADMINISTRATIVE Procedure) must be in
CONTROLS place
 There must be
established and
maintained system for
emergency response
- devices worn by workers to
protect them against chemicals
PPE toxins and pathogenic hazards
in the laboratory
 Least effective

AUTOCLAVE
- GEOBACILLUS STEAROTHERMOPHILUS (Biological
indicator)
 rod-shaped, gram-positive
 thermophile: resistance to heat
 Biological indicator; test the success of
sterilization cycles
 Check water level – 3L (3,000 mL)

- Time depends on the nature of material to be sterilized

- The most dependable system available for the

decontamination of laboratory waste and sterilization of
glasswares, media and reagents

- MOIST HEAT – irreversible coagulation and

denaturation of enzyme and proteins
 Temperature: 121°C
 Pressure: 15 psi
TYPES OF AUTOCLAVES
1 Gravity displacement
2 Vacuum assisted
DECONTAMINATIO DISINFECTIO STERILIZATIO
N N N
- Reduce the level of - eliminates - completely
microbial nearly all eliminates
contamination pathogenic -ex. autoclave
microorganism
-less lethal
process than

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 Clinical Internship 1
Internship 1 - EACMED Transcribed by: AAA

sterilization

BIOLOGICAL SPILL RESPONSE

1 Types of spill
2 SOP for spill clean up
3 Immediate action taken by staff
4 Spill team (who and how to contact them)
5 Spill kit and location
6 Post exposure medical program
7 Reporting structure
- POSSIBLE CAUSE OF BSR
 Human
 Mechanical
 Natural
MAJOR MINOR
BIOLOGICAL SPILL BIOLOGICAL SPILL
- Any biological spill - Any biological spill
occurred outside the BSC occurred within the BSC
(regardless the quantity)
BIOLOGICAL SPILL KIT CONTENTS
1 Container – for PPE
PPE
 2 pairs face shield
 4 pairs nitrile gloves
2  2 facemask and n95 respirator
 2 pairs shoe covers
 2 pieces head cover
 2 disposable gowns and coveralls
Absorption materials
 Towels
3
 paper towels
 spill pads
Clean up tools and materials
 Small disposable broom with dust pan and
wiper
 Biohazard waste bags
4  Forceps and tongs
 Concentrated bleach
 Bottle with water (for preparation of fresh
10% bleach)
 70% alcohol
5 Biological spill kit checklist
6 Biological spill kit response SOP
7 “DO NOT ENTER” signage
NOTE:
- Report all spills to biosafety officer and laboratory
supervisor

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Specimen preservatives and anticoagulants

TOPIC 2: QUALITY ASSURANCE AND PHASE OF 4  Ex. Sodium fluoride, Formalin, Boric Acid,
ANALYTICAL TESTING EDTA
Speaker: Sir Mark Kirby Baladiang, RMT 5 Specimen transport, processing, storage
PATIENT RELATED VARIABLES
INTRODUCTION - In preparing a patient for phlebotomy, care should be
- An effective testing process requires integration of taken to minimize physiologic factors related to activities
 Pre analytical that might influence laboratory determinations
 Analytical 1 Diurnal variation
 Post analytical
2 Exercise (ex. Creatinine)
- entire process of managing a sample must be Fasting (Low albumin for non-ambulatory or bedrest
3
considered patients)
 Sample collection = beginning 4 Ethanol Consumption
 Reporting and saving results = end 5 Tobacco smoking
 All process in between 6 Drug ingestion
7 Posture
- Essential to all health care aspects lab results that are
 Accurate
 Reliable
 Timely
PATH OF WORKFLOW

1. PRE-ANALYTICAL PHASE
- complex steps that must take place before a sample can
be analyzed such as test ordering, and sample collection

-major source or RESIDUAL “ERROR” and/or variable

that can affect test results
 Exercise = hyaline cast
 Diet = serotonin
 Posture
 DIURNAL VARIATIONS
o High in morning (ACAI) = I drink ACAI
tea during the MORNING
 ACTH
 Cortisol
 Aldosterone
 Iron
o High in PM (TPAG)
 Growth Hormone
 PTH
 TSH
 Acid Phosphatase
PRE-ANALYTICAL FACTORS
Patient-related variables
1
 Ex. Diet, age sex (GFR, crea, etc.)
2 Specimen identification 10 COMMON ERRORS IN SPECIMEN COLLECTION
3 Specimen collection and labelling 1 Misidentifcation of patient
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 Clinical Internship 1
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2 Mislabelling  requires rapid communication with a health care

3 Short draw/ wrong anticoagulant/blood ratio provider who can provide necessary medical
4 Mixing problems/ clots interventions
5 Wrong tubes/ wrong anticoagulant
- After releasing the accurate results at the right time, lab
6 Hemolysis/ lipemia results then are RECORDED in the designated logbooks
7 Hemoconcentration from prolonged tourniquet time and retained following the appropriate retention period of
8 Exposure to light/ extreme temperatures documents.
Improperly time specimens/ delayed delivery to
9 - Result interpretation is done by the physician/doctor.
laboratory
Processing errors; Incomplete centrifugation,
**NOTE DI KO SURE TO NASA NOTES KO LANG EWAN
10 incorrect log in, improper storage
KO RIN MAS MAGANDA UNG TABLE SA LAB MAGBASE
 Ex. K – cannot be recentrifuged (false ↑)
^_^
EXAMPELES OR PRE-ANALYTICAL FACTORS THAT
POSSIBLE POSSIBLE
MAY AFFECT CHEMISTRY RESULTS TEST LOW HIGH
EFFECT EFFECT
Bilirubi - -
n
<125 Dehydratio >125 Edema
mmol/ n mmol/ Hypervolemi
Na
L Vascular L a
collapse Heart failure
<2.5 >6.5
K mmol/ mmol/
L L
- 773 Acute urate
mmol/ nephropathy
Uric
L w/ tubular
Acid
blockage
Renal failure

2. ANALYTICAL PHASE 3. POST ANALYTICAL PHASE

- Considered as the ACTUAL LABORATORY TESTING or
the diagnostic procedures, processes, and products that
ultimately provides results, such as running a sample on an
automated analyzer
- QUALITY CONTROL in the laboratory is an essential part
of this phase as it involves the systematic monitoring of
analytic processes to detect analytic errors that occur
during analysis and to ultimately prevent the reporting of
incorrect patient test results.
-TASK
 Sample introduction and transport to cuvette or
dilution cup
 Addition of reagent
 Mixing of sample and reagent
 Incubation
 Detection
 Calculations
 Readout and results reporting
ASSESSMENT OF ANALYTIC CORRECTNESS OF
RESULTS
-to prevent the release of erroneous results, most
laboratories utilize a variety of flags or alarms
 Flags for Problem specimens
 Flags for Specimens that require additional
analysis w/ another method
 Flags for problematic results
- DELTA CHECK
CRITICAL VALUES
- Panic or alert value
- May represent a life-threatening situation that may not
otherwise be readily detectable
- Comprises result reporting and result interpretation
- Lab results must undergo a two-step post- analytical
review for:
 Analytical correctness – delta checks, linearity
ranges etc.
 Clinical significance – for the patient (applying
critical values, reference ranges)
*Importance of Assessment of Analytic Correctness
- TO PREVENT THE RELEASE OF ERRONEOUS
RESULTS
QUALITY ASSURANCE
-QUALITY MANAGEMENT SYSTEM (QMS)
 all aspects of the laboratory operation need to be
addressed to assure quality
- comprehensive set of policies, procedures, and practices
that are followed to ensure that a lab results are reliable
-encompasses a range of activities that enable lab to a
achieve and maintain high levels of accuracy proficiency
despite change in test methods and the volume of specimen
tested
 way of preventing mistakes/issues/problems
- A GOOD QA SYSEM
 SOP
 administrative eq such as mandatory record
keeping, data evaluation, and internal audit
-monitors QUALITY CONTROL (QC) program
-EQAS (External Quality Assessment Schemes)
GOOD QA SYSTEM
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- Establish Standard Operating System (SOP)for each step  Mean = average

of the laboratory testing process from specimen handling to  Mode = most common value
instrument performance validation.  Median = middle value
-reproducibility
- Specifies corrective actives, documentation, and the
person responsible for carrying out corrective actions when -produce series of result on the same
problems are identified. sample that agrees with each other;
consistency of results
- Monitors a Quality Control (QC) program.
-statistical measure of precision
- The lab must be accredited by an appropriate agency.  Standard deviation = ↑ SD, ↓
This gives official approval and states that the lab follows all precision
the guidelines set by the accrediting agency. Accrediting  Coefficient of variance =
agency will send inspectors to visit the lab view the records PRECISION ↑CV, ↓ Precision
and documents.

- Sustain high-quality employee performance

QA PROCESS
Identify issues with process

Generate Corrective Actions

-substance having a known determined

Verify Corrective Actions CONTROLS
range or values
-detect small concentration of a
substance
Implement Corrective Action  positive result
SENSITIVITY
- TIP
Monitor And Control  analytical = analyte
 diagnostic = patient/
P D C C A population
PLAN DO CONTROL CHECK ACT -ability to measure only the substance
of interest
QUALITY CONTROL SPECIFICITY
- also called STATISTICAL PROCESS CONTROL - ANALYTICAL SPECIFICITY =
 part of QUALITY ASSURANCE desired for confirmatory test
-an ideal concept which cannot be
TRUE VALUE
-process to periodically examine a measurement achieved
procedure to verify that it is performing according to pre- -the value approximating the true
established specifications ACCEPTED value, the difference bet. the two value
 Daily: frequency depends on the quantity of the TRUE VALUE is negligible
test performed each day (e.g., Hema – 2x a day)
 Patient Control can be used obtained from morning -the discrepancy between the result of
ERROR a measurement and the true (or
- refers to the measures that must be included during each accepted value)
assay run to verify the test is working properly. GOAL OF QC
1 to detect significant errors rapidly
- Detects significant errors rapidly. 2 report out good results in a timely manner
3 be cost effective and simple to use
- Simply to ensure that the results generated by the tests 4 if there is an error, identify the source of error
are correct *HOW MAY OR OFTEN?
- there should be at least one control per reportable result
- Ensure that a particular test method is working properly  reportable results include positive and negative
and that the test results are reliable CONTROLS
CALIBRATOR VS CONTROL INTERNAL EXTERNAL
CALIBRATOR CONTROL - built into the test device - consist of sample of
- reference material w/ - sample similar to human  Ex. control line in a known value
known concentration of specimen pregnancy test
analyte
EXTERNAL QUALITY ASSESSMENT (EQAS)
DEFINITIONS - refers to the system in which the performance of lab is
ACCURACY -agreement of results w/ true value for assessed PERIODICALLY AND RETROSPECITIVELY by
substance in given specimen an independent external agency
 College of American Pathologists = gold standard
-statistical measures include  INTERLAB
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INTERNAL QUALITY CONTROL  Sudden failure or change in the light source

- Inside the laboratory: shall be run daily and every time  Change in reagent formulation
the tests run in case of infrequently used tests  Change of reagent lot
 INTRALAB  Major instrument maintenance
 Sudden change in incubation temperature
- IQC FOR QUANTITATIVE RESULTS (Enzymes only)
 Levy Jenings  Change in room temperature or humidity
o Should be used to plot daily QC values.  Failure in the sampling system
o It also indicates the changes in trends  Failure in reagent dispense system
and shifts in the lab performance  Inaccurate calibration / recalibration
 Westgard Rules
o used to interpret daily QC values WESTGARD RULES
 1S = expected result (normal)
 13s = you have to do something
 2s = accepted result for manual
SYSTEMIC VS RANDOM
SYSTEMATIC RANDOM
- inaccurate - imprecise
 Measured by MEAN  Measured by SD
1. SYSTEMATIC ERROR
- It is evidenced by a change in the mean of the control
values.
 TREND
 SHIFT
A. TREND
– gradual shift, continuously increasing or decreasing
values. (6 or more results)
 values for the control chart that continue to either
increase of decrease over a period of 6
consecutive days
 Gradual loss of reliability in the test system
 Usually subtle

**NOTE: all EVEN numbers are SYTEMATIC ERROR

-CAUSES
 22s, R4s, 41s, 10x
 Deterioration of the instrument light source
 Gradual accumulation of debris in sample / reagent
NATIONAL REFERENCE LAB
tubing
Lung Center of the
 Gradual accumulation of debris on electrode Clinical Chemistry
Philippines
surfaces
National Kidney Institute Hematology (CBC)
 Aging of reagents
Microbiology: sputum for
 Gradual deterioration of control materials DSSM
 Gradual deterioration of incubation chamber Research Institute for
temperature (Enzymes only) Tropical Medicine
Blood Bank: Transfusion
 Gradual deterioration of light filter integrity Transmissible Infections TTIs
 Gradual deterioration of calibration East Avenue Medical
B. SHIFT Drug Testing
Center
– abrupt changes, sudden rise or fall from the mean values STS/ AIDS Cooperative
 6 or more consecutive daily values that distribute Central Laboratory STIs
themselves on one side of the mean value line, but (SACCL)
maintain a constant level.
 Shifts in QC data represent a sudden and dramatic
positive or negative change in test system
performance

TOPIC 3: PHLEBOTOMY
Speaker: Jason Paul V. Sarian, RMT

PHLEBOTOMY
-CAUSES -derived from Greek word
 Phlebo = vein
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 Tomy = cutting 3 Winged infusion set (Butterfly needles)

4 Tourniquet
- is the incision of a vein for the purpose of collecting 5 Gloves, Gauze pads, Alcohol, puncture container
blood 1. EVACUATED TUBE SYSTEM
- made up of
PHLEBOTOMIST  Adapter/holder
- is the one performing blood collection  Color-coded/ evacuated tubes
 Double-headed needle/2-way needle
- must obtain
 High school diploma -small tubes are available for PEDIATRIC and
 Phlebotomy training via hospital, technical GERIATRIC collections
school (MedTech), or training modules 2. SYRINGE
 Employers may require phlebotomy - most commonly used to obtain blood
certification via national certification 3. WINGED INFUSION SETS (BUTTERFLY NEEDLES)
examination - can be used when blood has to be collected from a very
o PBCC small vein
DUTIES  21, 23, 25
To assist the healthcare team in the accurate,
1 safe, and reliable collection and transportation of SPECIMEN COLLECTION & HANDLING
blood/specimens for processing/testing - most common specimen collection methods
2 Identify patient correctly 1 Skin puncture
3 Assess patient before and after blood collection 2 Venipuncture
4 Other function: Blood donor phlebotomy
OTHER RESPONSIBILITIES: SKIN PUNCTURE
*COMMUNICATION SKILLS - should be used in
- FACE TO FACE communication is the most important  INFANTS and NEWBORN = smaller total
and effective form of communication and is part of volume of blood
phlebotomist’s job everyday o Venipuncture can cause hospital-
 Verbal induce anemia
 Non-verbal  Adults
 Professional Appearance o obese,
1. VERBAL COMMUNICATION o with burns,
- use simple and everyday vocabulary o small or severely damaged veins
- avoid use of slang  IV fluid is flowing into the only accessible vein
- avoid sarcasm
 Save veins for patients receiving chemotherapy
- use calm soothing confident tones
 Elderly
2. NONVERBAL COMMUNICATION
- show a pleasant facial expression
- mixture of capillary, venous, and arterial blood
- face to face positioning
- smiling
- Lab values varies from venous specimens
- erect posture
 Lower (↓)
- eye contact
o RBC count
3. PROFESSIONAL APPEARANCE
o Hct
- good posture
- grooming o Hgb
 neatly combed hair o Plt count
 clean fingernails
-LANCET LENGTH = 1.75mm (preferred)
 pressed uniform
 proper make up
- DEPTH OF INCISION
 proper attire
 Infants/ Children: <2.0mm
 Socks = long enough to cover skin
 Adults: <2.5mm
 Shoes = 1 ½ inch heels are standard o Thicker sites
GUIDE TO A SUCCESSFUL PHLEBOTOMY
“A successful phlebotomy starts when you are able to
VENIPUCNTURE
identify and access most suitable vein”
PROCEDURE
 Prepare = prepare yourself, replay the
Patient interaction
procedure in your mind
 Identify patient
 Suitable vein selection
 Note patient isolation requirements
 Proper technique
1  Note patient dietary restrictions
 Practice
 Reassure patient
 Do not be nervous
 Verify paperwork
 Establish rapport with your patient
 Position patient
2 Assemble supplies and equipment
BLOOD COLLECTION DEVICES
3 Apply tourniquet
1 Evacuated tube system
4 Palpate the vein
2 Syringe
5 Apply antiseptic
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 Antiseptic for Blood C/S: alcohol  iodine

 alcohol
6 Insertion of needle
7 Release tourniquet
8 Completion of injection
Application of sterile pad prior to withdrawal of
9
needle and syringe
Specimen preparation
 If syringe is used, fill tubes
1
 Discard needle
0
 Label specimen
 Transport specimen promptly and properly

REASON FOR SPECIMEN REJECTION

1 Hemolysis/Lipemia
2 Clots present in anticoagulant specimen
3 Non-fasting specimen when test require fasting
4 Improper blood collection tube
5 Short draws, wrong volume
Improper transport condition
6
 Ice for blood gases
Discrepancies between requisition and specimen
7
label
8 Unlabeled or mislabeled specimen
9 Contaminated specimen / leaking

ORDER OF DRAW FOR MULTIPLE TUBE

COLLECTIONS
COLLECTION
COLOR INVERSION
TUBE
Yellow Blood culture – SPS 8-10 times
Blue Citrate tube 3-4 times
5 times (plastic)
Serum tube (glass or
Red
plastic)
None (glass)
Serum Separating
Gold 5 times
Tube
Green Heparin tube 8-10 times
Purple EDTA tube 8-10 times
Fluoride (glucose)
Gray 8-10 times
tube

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 Clinical Internship 1
Internship 1 - EACMED Transcribed by: AAA

S – surgical/ C-cytologic
TOPIC 4: BASIC HISTOPATHOLOGIC TECHNIQUES
Speaker: Sir Ceasar Abigania, RMT

HISTOPATHOLOGY
- concerned with the diagnosis of abnormalities within
tissues and cells by microscopic examination
 HISTOLOGY = normal cells
 HISTOPATHOLOGY = abnormal cells SCHEDULING OF FROZEN SECTION, FNAB, &
ULTRASOUND/CT SCAN GUIDED BIOPSIES
-etymology - scheduled at least 2 days prior to procedure
 Histo = tissue
 Pathos = suffering - FNAB, CT/Ultrasound guided biopsy = performed by
PATHOLOGIST
 Logos = study
1. FIXATION
SPECIMEN COLLECTION, PRESERVATION,
- The process of fixation can produce ARTIFACTS in the
RECEIVING, AND LABELLING
tissues if the procedure is not carried out under optimal
A. SPECIMEN COLLECTION & PRESERVATION
conditions
- TYPES
 Purpose: to prevent autolysis
 Routine Histological Examination
 Most critical step
 Frozen Section
 Routine Non-Gynae Cytology - AIM
A. ROUTING HISTOLOGICAL EXAMINATION  preserve morphologic and chemical integrity
- all specimens for routine histological examination must of cell = primary
be collected into containers of FORMALIN solution o Done by stabilization of protein
 10% NEUTRAL BUFFERED FORMALIN
 harden and protect tissue from trauma =
secondary
- there must be sufficient formalin in the container to
completely cover the specimen
- If fixative does not have proper access to the tissues or
because of the nature and quality of the reagent used
- Specimen must NOT be placed into another solution or
FORMALIN PIGMENT
into a dry container
- This pigment appears as brown to black, finely granular
B. FROZEN SECTION
associated with, or in the vicinity of RBCs
- Specimens for frozen section is processed w/o
preservative and must be placed in a suitably sized dry
- most often seen in tissues rich in blood such as spleen
specimen container and transferred immediately to the
and in tissues w/c have had prolonged fixation
Histopathology section
 TAT = 15mins - artifact can be removed by treating the section w/
C. ROUTINE NON-GYNAE CYTOLOGY SATURATED ALCOHOLIC PICRIC ACID solution prior
- must be freshly collected and placed in a sterile to staining or prevented by using BUFFERED
container FORMALIN solution and restricting fixation time
 Papsmear, pleural fluid
B. SPECIMEN RECEIVING, LABELLING, &
ACCESSIONING 2. GROSSING
- All histopathology specimens shall be received and - brain = fix  grossing (Exception)
logged at the histopathology section during its operation
hours - Check for SSCC

- All specimens must be properly and legibly labelled with *S- SIZE
the ff: - 3D measurement (in cm)
 Patient’s name,  Length
 Age and sex  Width
 date & time of collection,  Height
 type of specimen
 Accession number -2D (cm)
 Length
- All specimens must be properly logged and  Width
accessioned in the corresponding receiving logbook
 Surgical Logbook *S-SHAPE
 Cytology Logbook - Tubular
o Pap smear  Appendix
o Aspirates  Gallbladder
- Irregular
- The request form and the specimen container shall be A – typical appearance
labeled with the assigned accession number *C-COLOR associated with RBCs in
- Grayish-Tan kidney section

B- Pigment in a Page
section12ofof 20
fatty liver where it is also
deposited in the vicinity of
RBCs
 Clinical Internship 1
Internship 1 - EACMED Transcribed by: AAA

- Brownish-red, etc - remove the appendix when it is infected

 Shape: tubular
*C-CONSISTENCY
- soft, rubbery* -TANGENTIAL SECTIONING = grossing of appendix
 3 blocks
 Include fats = take note if present
TOTAL ABDOMINAL HYSTERECTOMY AND
BILATERAL SALPHINGO-OOPHORECTOMY -VERMIFORM = U-shaped
(TAHBSO)
- removal of ENTIRE uterus, ovaries, fallopian tube, TISSUE PROCESSORS:
and cervix LEICA TP 1020
 Hysterectomy = removal of uterus only 12 steps (1 step per hour) = 12 hours
 Salpingo = fallopian tube 10% Neutral Buffered Formalin
o 3 blocks 1
- prevent crystalline pigment
 Salpingo – oophorectomy = both fallopian tube & 2
ovaries 3 95% Alcohol (3x)
 Oophorectomy = ovaries 4
o 1 block 5
o Cervix = 2 blocks (L & R) 6 100% Alcohol (3x)
7
-large radical 8
 w/ fluid = large radical w/ cell cytology 9
Xylene (3x)
1
-take note if CYST is present 0
1
MODIFIED RADICAL MASTECTOMY (MRM) 1
- A procedure in which the entire breast is removed Paraffin Wax (2x)
1
including the skin, areola, nipple and most axillary 2
lymph nodes
 Pectoralis major is spared 3. DEHYDRATION
- removing of extracellular and intracellular water from
- Surgery for breast cancer tissue following fixation and or prior to wax infiltration
 Alcohol = most common
- 3D GROSSING MEASUREMENT o Ethyl = best
 Axillary tail (lymph nodes) w/ nipple or areola o Isopropanol = best substitute for ethyl
GALLBLADDER LAPAROSCOPIC o Butanol = plants
CHOLECYSTECTOMY o Tertiary butanol = universal solvent
- also known as MINIMALLY INVASIVE
o Methanol = toxic
CHOOLECYSTECTOMY
 Acetone = rapid, cheap but extremely volatile
- performed through 4 small incisions with use of a o Fixative and dehydrant
camera to visualize the inside of the abdomen and long  Cellosolve = combustible
tools to remove the gallbladder  Dioxane = excellent dehydrating and clearing
 3-4 blocks = choose section w/ abnormality (ex. agent, can explode
darkening) o Universal solvent
o Faster = 25x
-GROSSING INCLUDES  Tetrahydrofuran = dehydrates and clears
 Measurement of thickness of wall tissue, toxic
 Define mucosa, serosa o Universal solvent
o Mucosa = inner
o Serosa = outer 4. CLEARING
o Velvety, smooth - removal of alcohol from section immersing them in
 Note if gallbladder is already open ante-medium or DEALCOHOLIZATION
o Stone = hardness, color, size (smallest
to largest) -makes tissue TRANSPARENT
 Shape: Tubular  Xylene (Xylol) = most common
TOTAL THYROIDECTOMY o Enhance tissue color
- removal of the entire thyroid gland  Chloroform = tough tissues
 Benzene = rapid acting, prolonged exposure
- often performed to treat thyroid cancer, but it may also lead to bone marrow damage  APLASTIC
be performed to treat uncontrollable hyperthyroidism or ANEMIA
goiter that causes severe symptoms  Toulene = substitute for xylene and benzene
 Essential oils
-section by o Cedarwood oil = CNS
 QUADRANTS (4) o Clove oil = expensive
 Center (isthmus) = helical o Aniline oil = delicate specimen (ex.
APPENDIX APPENDECTOMY embryo)
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 Carbon tetrachloride = same w/ chloroform but -method for the preparation of thin sections for material
cheaper such as bones, minerals, and teeth, and an alternative to
 Methyl benzoate = double embedding electopolishing and ion milling
 Mikros = small
5. INFILTRATION/ IMPREGNATION  Temnein = to cut
- process of complete removal of clearing reagents by
substitution of paraffin or any such similar media such as WATER BATH = 45-50 C
beeswax  Depend on infiltrating medium = 6-10 degrees
below melting point of infiltrating medium
-cleared tissue is infiltrated w/ wax (usually paraffin) which  2 segments in 1 slide
is liquid at 60 C and then allowed to cool to 20 C in order  Albumin, plasma, serum = for ribbon to adhere
to solidify into a consistency that allows sections to be cut more effectively on slides
o Not applicable for IHC
-CARBOWAX = aqueous impregnation medium  Positive slide = natural adhesive
 Fixation to directly impregnation
OVEN = 2-5 degrees above
6. EMBEDDING, CASTING, AND MOLDING  Too high = fry, pyknotic
- process by w/c tissues are surrounded by a medium  Too low = paraffin will not melt
such as agar, gelatin, or wax w/c when solidified will
provide sufficient external support during section Panel
- Preferred: PARAFFIN WAX -thickness (4-6 micron)
ORINETATION –
15 degrees = optimum angle
-SUBSTITUTE FOR PARAFFIN WAX  Clearance angle = 3-8
 Paraplast (MP 56-57 C) = synthetic polymer  Bevel angle – 27-32
o Stronger than paraffin
o Used in the laboratory 10. STAINING
 Embeddol (MP 56-58C) – less brittle than - Process whose purpose is to optically differentiate the
paraplast cell, cellular and tissue constituents by variation in color.
 Bioloid – for ice ROUTINE HISTOPATHOLOGY STAIN
 Tissue mat – REAGENT TIME/DIPS
 Ester wax (MP 46-48C)– most commonly used 1
Xylene (2x) 5 minutes
 Carbowax = no need for dehydration and 2
clearing 95% ethyl alcohol
o Dissolve in water bath 3 - rehydration 5-10 dips
 Celloidin – for frozen section/ CNS specimen  Stains are aqueous dyes
Running H2O
4 5 dips
7. BLOCKING - for specimen to become aqueous
- remove cassette from mold HEMATOXYLIN
-nuclear stain
5 20 minutes
-block is now read for section (microtomy)  Nuclei = red (Temporarily)
 Organelles/ cytoplasm = blue
8. TRIMMING Running H2O
6 5-10 dips
- removing excess wax from block, so that it forms a 4- -remove stain
sided prism or until it assumes the shape of a truncated Acid Alcohol
pyramid opposite side being parallel 7 - decolorizer 1 dip
 removes stain in cytoplasm
-at least 2mm of wax should surround the tissue block 8 Running H2O 5-10 dips
 COARSE trimming = sides, tips, bottoms are Ammonia
trimmed 9 2 dips
- bluing agent (blue)
 FINE trimming = to expose tissue 1
Running H2O 5 dips
0
9. CUTTING EOSIN Y
-TYPES OF MICROTOME - cytoplasmic stain/ counter stain
 1
Rotary = paraffin-embedded (most common)  Y = yellowish 5-10 dips
1
o 4-6 micra  B = bluish
o Used in the laboratory  YB = yellowish-bluish
 Cryostat = 4-6 micra 1 95% ethyl alcohol
5-10 dips
 Rocking = large blocks (10-12) 2 - differentiate cytoplasm/ organelles
 Sliding = celloidin (4-9 micra) 1
o Most dangerous type 3 Xylene (2x)
2 minutes
 Freezing = unembedded tissue section (fresh) 1 - intensify refractive index of tissue
 Ultra thin = 500 – 1,200 4
 Semi thin = 0.5 – 1 microns 1
Blow dry
5
MICROTOMY 1 Labelling

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6 o Breast, etc = -15

1
Mounting Coverslip
7 - The rapid freezing of the tissue sample converts the
FROZEN SECTION water into ice.
REAGENT TIME/DIPS  The firm ice within the tissue acts as embedding
1 Hematoxylin 1-2 minutes media to cut the tissue.
2 Running H2O 5-10 dips  Medium = ISOPENTANE
3 95% ethyl alcohol 5-10 dips
4 Running H2O 5-10 dips - The frozen section is mainly used for rapid diagnosis of
5 Eosin Y 1-2 dips the lesion for intraoperative management, to know the
6 extent of the lesion, to do enzyme immunocytochemistry
95% ethyl alcohol 5 dips (2x) and immunofluorescence study and also to stain lipid and
7
8 Blow dry certain carbohydrate in the tissue
9 Labelling FROZEN SECTION PROCESSOR: LEICA 1860
1 PURPOSE:
Mounting coverslip 1 Rapid pathologic diagnosis during surgery
0
PAPSTAIN (PAPSMEAR STAIN) 2 Diagnostic and research enzyme histochemistry
1 Hematoxylin 5 minutes Diagnostic and research demonstration of soluble
3
2 Running H2O 5-10 dips substances such as lipids and carbohydrates
3 Ammonia 1-2 dips Immunofluorescent and immunohistochemical
4
4 Running H2O 5-10 dips staining
5 Orange G-OG6 5 dips Some specialized silver stains, particularly in
5
6 neuropathology
95% ethyl alcohol 5-10 dips (2x) TISSUE FREEZING
7
8 Eosin Azure 50 30 seconds - The tissue for freezing should be fresh, and freezing
should be done as quickly as possible.
9
1 95% ethyl alcohol 5-10 dips (2x)
- Slow freezing can cause distortion of tissue due to ice
0
crystal artifacts.
1
Blow dry COMMONLY USED METHODS OF FREEZING
1
1 Liquid nitrogen
1
Labelling 2 Isopentane cooled by liquid nitrogen
2
3 Carbon dioxide gas
1
Mounting Coverslip 4 Aerosol sprays
3
IODINE
- Used to identify pre-cancerous and cancerous changes
in cervical and vaginal tissues during "Pap smear" follow
up examinations in preparation for biopsy.

- The acetic acid causes the abnormal cells to blanch

white, while the normal tissues stain a mahogany brown
from the iodine.

- AKA “OLDEST STAIN

11. MOUNTING
- (2ND to the) Last procedure in the series that ends with
a permanent histological preparation on the table, well
after the tissue processing and staining.

- 2 TYPES OF MOUNTING MEDIA:

 Aqueous – Temporary
 Resinous – Permanent
o 1.5 refractive index

12. LABELLING
- Most (last) important step in histopathology technique

FROZEN SECTION
The frozen section is the rapid tissue section by cooling
the tissue with the help of CRYOSTAT to provide
immediate report of the tissue sample.
 CRYOSTAT is the instrument to freeze the
tissue and also to cut the frozen tissue for
microscopic section.
 Temperature: -20 C

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 Clinical Internship 1
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3 Hematocrit (Hct)
4 WBC count
5 WBC Differential Count
6 Platelet Count
ANTICOAGULANTS
Color Action Test
Routine
EDTA Lavender Chelates Ca hematology
(CBC)
Binds w/
Coagulation
calcium
Trisodium Test (1:9)
Blue
TOPIC 5: ESSENTAILS OF COMPLETE BLOOD citrate
Preserves RBC
COUNT D-dimer
morphology
Speaker: Ma’am Ray-Ann, RMT Osmotic
fragility of
ESSENTIALS OF CBC erythrocytes
1 Pre-examination and Post-examination Phase Binds and
(OFT)
2 Technical Procedures Heparin Green enhance action
Quality Assurance in CBC and Quality Control of anti-thrombin
3 Immuno-
Procedures phenotypin
g
PHASES OF EXAMINATION *NOTE
1. PRE-EXAMINATION - Clotted blood specimen are UNACCEPTED (False ↓)
- includes specimen handling issues that occur even prior
to the time the specimen is received in the laboratory. ORDER OF DRAW
“Stop Light Red, Stay Put Green, Let’s Go”
- Important errors can occur during this phase with COLOR COLLECTION TUBE INVERSION
specimen and identification Yellow
(PEDIA)
-VARIABLES Blood culture – SPS 8-10 times
Green
 Selection of assay relative to patient need (Adults)
 Implementation of assay selection Blue Sodium Citrate tube 3-4 times
 Patient identification and preparation Serum tube or gel
 Specimen collection, equipment, and technique separator None (glass)
 Specimen transport, preparation, and storage Red
 Monitoring of specimen condition (Plastic = w/ clot 5 times (plastic)
2. EXAMINATION activator)
-VARIABLES Green Heparin tube 8-10 times
 Centrifugation of red top tubes for electrolyte Purple EDTA tube 8-10 times
tests Fluoride (glycolytic
3. POST EXAMINATION inhibitor) tube
Gray 8-10 times
- The final phase of the laboratory process.
ESR
- This phase culminates in the production of a final value,
result, or in the case of histology, a diagnostic pathology EFFECT OF STORAGE ON BLOOD COUNT
report. - stable for 8-24 hours when stored at 4 C

-VARIABLES - best to count WBC and platelets w/n 2 hours of

 Accuracy in transcription and filing of results collection
 Content and format of laboratory report, narrative
report - Reticulocytes are stable for 4 C or 24 hours but should
 Reference interval and therapeutic range be counted w/n 6 hours at RT
 Timeliness in communicating critical values
 Patient and physician satisfaction - Hgb and Hct may decrease w/n 2-3 days
 Turnaround time STABILITY OF CBC PARAMETERS
 Cost analysis ROOM TEMP REF TEMP
PARAMETERS
 Physician application of laboratory results (18-25 C) (4 C)
Hemoglobin
N/A 72 H
RBC Count
CLINICAL HEMATOLOGY
CV (Hct) 1-4 H 6-12 H
*HEMATOLOGY
- The branch of medicine concerned primarily with Retic Count
N/A
studying the formed elements of blood (blood cells) and Platelet
the blood forming tissues WBC count 24-72
COMPONENTS OF CBC Automated Differential 6H
1 RBC count Count
2 Hemoglobin (Hgb) Peripheral Blood Smear <3H 8H

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- PRINCIPLE
MATERIALS USED  Whole blood is diluted with a weak acid solution
1. HEMACYTOMETER: NEUBAUER COUNTING to lyse the red cells and facilitate counting of
CHAMBER leukocytes
- is a technique used to enumerate the total cell count in
the blood or other biological body fluids -SPECIMEN

- used when verifying RBC, WBC, and platelet count

- PRINCIPLE
 blood is diluted with appropriate known volume
of diluting fluid and then counting is done by
using hemocytometer

- PURPOSE OF DILUTING FLUID

 To lyse/ destroy RBC, WBC or platelet when not
needed such as when reading WBC count, it will
lyse the RBC to make only WBC visible
o WBC = 1:20
o RBC = 1:200
o Platelet = 1:100

2. THOMA PIPETTE
- Consists of graduated capillary
rube, mixing bulb with glass bead
and aspirating tube
 RBC and PLATELET =
101th mark
 WBC and SEMEN = 11th mark
3. DILUTING FLUIDS
A. GLACIAL ACETIC ACID – WBCs
- Destroys (lyses) RBCs and platelets

– One of the compositions of Turk’s Fluid (gentian violet):

 Stains the nuclei of WBC
B. HAYEM’S SOLUTION – RBCs
- components
 Sodium chloride:
o maintains osmolarity and provides
isotonicity, so that red cells maintain
their shape and size
 Sodium sulphate:
o prevents aggregation of RBCs; it also
acts as an anticoagulant and fixative.
 Mercuric chloride
o acts as a preservative
C. REES-ECKER / 1% AMMONIUM OXALATE –
PLATELET
- components
 Brilliant Cresyl Blue: stain
 Sodium citrate

4. TALLY COUNTER
- for counting cellular elements

WBC COUNT
MANUAL METHOD

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 Clinical Internship 1
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 EDTA WHOLE BLOOD PROCEDURE

PROCEDURE Fill about 3⁄4 of a capillary tube with blood from a
1
Aspirate the blood specimen up to 0.5 mark of the finger puncture or an EDTA tube
1
Thoma pipette Plug the other end of the tube
Quickly dip the pipette into the diluents then aspirate 2  (i.e., the end that has not come in contact
2
to 11th mark with the blood with soft wax or plastic)
3 Shake for 2 minutes then discard a few drops Place the capillary tubes in centrifuge. The sealed of
3
Charge the counting chamber and allow the cells to the tube should point outwards away from the center
4
settle for 1 minute before counting (wait cells to settle) Centrifuge at 10,000 G for 5 minutes
4
5 Count the cells  3,000 G for 10 minutes
FOCUSING 5 Read
*WBC
 10x magnification (LPO) HEMOGLOBIN
 Count 4 large corner squares - SAHLI’S ACID HEMATIN METHOD
 Obsolete
*RBC/PLT - PRINCIPLE
 40x magnification (HPO)  Hemoglobin present in the
 Count Large center square RBCs is converted to acid
o 4 corners and center hematin which is dark brown
medium square colored compound

-SAHLI’S HEMOGLOBINOMETER 
COMPUTATION
*RBC and WBC count
Cells counted X DF X volume correction factor PROCEDURE
1 Place N/10 HCl into Hb tube up to 2 gram
Volume of 1 square X No. of squares 2 Blood sample in Sahli’s Hb pipette up to 20 uL
3 Add blood sample to acid solution
*VOLUME CORRECTION FACTOR 4 Mix with a stirrer
5 Allow to stand for 10 minutes
*TOTAL CELL COUNT Add distilled water drop by drop till the color of the
6
solution matches to brown glass standard
N × Dilution Factor 7
Take the reading of the lower meniscus from the
graduated tube in grams
Depth ( 0.1 ) × Area counted
RBC INDICES
Counting of RBCs by MANUAL METHOD is NOT -includes
RECOMMENDED due to poor reproducibility, and  Mean cell volume (MCV)
therefore, poor precision  Mean cell hemoglobin (MCH)
 Mean cell hemoglobin concentration (MCHC)
PACKED CELL VOLUME / ERYTHROCYTE VOLUME 1. MEAN CELL VOLUME
FRACTION (HEMATOCRIT – HCT) Average volume of the red blood cell (RBC)
- It is the volume percentage of RBCs in blood.
 When verifying hematocrit

- PARTS OF HCT
 Fatty layer
 Plasma
 Buffy Coat
2. MEAN CELL HEMOGLOBIN
 Packed RBC - average weight of hemoglobin (Hb) in the RBC
 Wax plug

3. MEAN CELL HEMOGLOBIN CONCENTRATION

(MCHC)
- Average concentration of Hb in the RBC volume

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-round shape, colorless but may

PERIPHERAL BLOOD SMEAR Lymphocyte appear as very light purple to pink
- Confirmation of the presence of abnormal blood cells color
 correlates the released blood cell counts from -large round or oval cell and variably
Monocyte
automated machines and blood cell counts from irregular
manual methods) Eosinophil 2-3, bright pink marks and granule
TWO-SLIDE (WEDGE) METHOD Deep purplish blue to dark purple-
Basophil
Get 2 clean glass slides. Place 1 small drop of red small
1
blood on the far end of the first glass slide. Neutrophil Pink or purple blue, mostly seen
Position the 2nd glass slide at 30 – 45° before the
2 drop of blood on the first glass slide to allow the
blood to spread out evenly on the vertex.
Move the 2nd glass slide rapidly and smoothly over DXH SERIES
3
the length of the1st glass slide at a constant angle. DXH 560
4 Air-dry the 1st glass slide - Quantitative, multi-parameter, automated hematology
5 Fix and Stain. Count a differential analyzer with an autoloader for in vitro diagnostic use in
CRITERIA OF A GOOD SMEAR clinical diagnosis
1 2/3 to 3/4 the length of the slide.
There should be a gradual transition from the thick
2 -PRINCIPLE
area on the thin area. Read at the monolayer area
must have a feathery edge that is very slightly  Coulter principle = WBC, RBC, Plt
3 rounded and has a RAINBOW appearance when
the slide is held up to the light.  Optical/ Impedance = Differential Count
ERROR/ UNACCEPTABLE PBS  Photometric = Hemoglobin
 Calculated = Hematocrit (calculated in RBC)
DXH 500 SERIES REAGENT
- Enhancement low formaldehyde
producing isotonic buffered
solution.
DILUENT
- Dilutes the specimen and is used
for rinsing module components
between sample analyses
A Chipped or rough edge on spreader slide – Cyanide-free lytic agent that
B Hesitation in forward motion of spreader slide lyses RBC for WBC count.
C Spreader slide pushed too quickly
D Drop of blood too small LYSE
– Classification of WBC
Drop of blood not allowed to spread across the subpopulations and hemoglobin
E
width of the slide (bullet shape) measurement
F Dirt or grease on the slide - Azide – free, formaldehyde – free,
Uneven pressure on the spreader slide (most biodegradable cleaner that
G
common error) CLEANER (blue) contains proteolytic enzyme that
H Time delay aids in the removal of protein
buildup.
RAPI-STAIN
- Used for staining bone marrow morphology, leukocytes
in blood film, and malarial parasites
STEPS
1 Fix air dried smears in Fixative solution 5 seconds
fixation.
2 Transfer the slide to Eosin and dip 5 times
3 Transfer the slide to Azure/Methylene blue 5 dips
4 Wash briefly in water and allow to dry
5 Examine under oil immersion lens. DXH CONTROLS and CALIBRATORS
A. CONTROL
- Tri-level integrated control that enables monitoring of the
system’s performance and calibration status for all CBC
and differential parameters

- 1 set is used for 15 days

 Blue: LOW
 Red: Normal
 Green: High
B. CALIBRATORS
- Traceable to reference methods and recommended for
determining the adjustment of the directly measured CBC
parameters.
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7 Store the control 2-8°C

- Calibration status should be monitored with Beckman 8 Validate the results on the QC icon.
Coulter controls 9 Log the results in the designated QC log sheet.
DXH 560 QUALITY CONTROL
1 Prepare the controls
Select Run Icon to display the sample
2
analysis- patient result screen.
3 Mix the controls
Fully insert the control into the cassette and
place the cassette in the input area by the
following arrows on the cassette for correct
placement. Then select Start Icon.
4  Note: Running control tubes with the
bar code facing away from the front
of the cassette window can cause the
control to be run as patient sample.
The bar code must be facing out
Remove the controls from the cassette when
5 the cassette reaches output area, and the
control is finished processing.
6 Store the control 2-8°C
7 Validate the results on the QC icon
8 Log the results in the designated QC log sheet

DXH 520 QUALITY CONTROL

- Sample Analysis:
 Closed-vial mode
 open- vial mode

- DAILY CHECKS: starts a series

of qc and checks to determine
whether the dxh520 system is
running properly.

PROCEDURES
1 Prepare the control
Select Display/Run Icon to display the sample
2 analysis-patient results screen. The door
automatically opens
Click next icon and use the bar-code scanner to the
3
control bar code and select Check Icon
4 Mix the control
Fully insert the control into the tube holder and select
5 Run Icon. The door automatically closes to initiate
the analysis.
Remove the control when the door opens, and the
6
control is finished processing.
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