Experiment 1:
Instrumentation in Serology
Activity #1
Various instruments and equipment are available
in the laboratory and many of this have similar
uses in the different areas of laboratory testing.
However, materials used for serology must be
maintain only for test undertaken in this section.
Proper handling, usage and care should be done at
all times since this also contribute as aspect in
quality assurance.
Objectives:
1. Identify correctly and describe the uses of
various glassware, instruments, and
equipment specifically in serology.
2. Demonstrate the proper handling and
care of these instruments.
Light instruments
1. ORDINARY TEST TUBES AND TEST TUBE
RACK
 Test Tubes
- Also known as sample tubes
- It is a common peace of
laboratory glassware consisting
of a fingerlike link of glass or
clear plastic tubing opened at the
top, usually with a rounded U
shape button.
- It is available in a multitude of
length and lids.
- The test tubes are used to hold,
mix, or heat small quantities of
solid or liquid chemicals,
especially for qualitative
experiments and assays.
- It may be a plastic or made of
glass.
 Test Tube rack
- Its support the test tubes
during experimentations.
- It’s serve as a holder of our
test tubes.
- Its hang the test tube upside
down for dry.
- It’s made of plastic, metal or
wood
2. GRADUATED CENTRIFUGE TUBE
- Are cylindrical shaecalradiplastic or glass
containers, they are design to fit into the
centrifuge slot for the analysis and
separations of various materials
- They separate solid out of liquid chemical
solutions.
3. PIPETTES
- Either Made up of plastic or glass.
- They are used to transfer unknown
volume of liquid from one container
to another. Usually for volumes of 20
mL or less than.
- They maybe reusable or disposable.
Pipette Classification
I. Design
a. To contain (TC)
- It holds or contains a particular
volume but does not dispense the
exact volume.
b. To deliver (TD)
- It dispense the volume indicated.
II. Drainage Characteristics
a. Blowout
- It has a continuous etched ring or
two small, close continuous rings
located near the top of the pipet.
This means that the last drop of
liquid should be expelled into the
receiving vessel.
b. Self-draining
- It drains by gravity
III. Type
a. Measuring or Graduated
- It is capable of dispensing
several different volumes
1. Serologic
2. Mohr
3. Bacteriologic
Disposabl
 4. Ball, Kolmer, or  Plunger Button
Kahn - It performs the following 2
5. Micropipette functions:
b. Transfer 1. Volume Adjustment
-
It is design to dispense one o Rotate the plunger
volume without further clockwise, if you
subdivisions. want to decrease the
1. Volumetric volume
2. Ostwald-Folin o Rotate the plunger
3. Pasteur pipettes counterclockwise, if
you want to increase
4. Automatic
the volume
macropipettes or
o A distinct click
micropipettes sound at every
1. Serological Pipette volume change
- Has etched rings insures perfect
volumes settings
- It is classified as measuring or and prevent any
graduated type accidental volume
- It is also a blowout pipette change.
- It has graduation marks down on 2. Liquid aspiration or
the tip, it is intended for the dispensing
delivery for pre deter mint volumes. o By press and
depress the
2. Automatic Pipette plunger to
- Also known as the micropipettes aspirate or
- They are used to accurately transfer dispense liquid.
small volume of liquids.
 Tip Ejector button
- Glasspirant are not highly accurate
for less than one ml but the - It allows safe, effortless and
micropipettes are both accurate and quick ejection of tips.
precise. - The tips can easily remove
from the micropipettes by
Parts of the Automatic Pipettes
pressing the tip ejector
Plunger Button button.
 Volume Display or digital
Tip Ejector volume indicator.
Volume Display Button
- Its shows the volume of the
liquid to be aspirated or
Shaft dispensed.
Tip Cone
 Shaft

 Tip cone
- It provide fitment to the tips.
  Disposable pipette tips
- There are 2 sizes of tips:
1. The larger BLUE tips–
They are used for the
P1000
5. To empty the tip completely press the plunger
to the second stop
6. Wipe the tip on the inner wall while taking the
tip out of the vessel

2. The smaller CLEAR tips – Plunger down first stop

They are used for P20 Then up again
and P200 Then release
Dispense, first stop down
Then second stop
And Release the plunger

2. Reverse Pipetting Technique

There are 2 types of pipetting techniques
1. Forward pipetting technique
2. Reverse Pipetting
1. Forward pipetting technique
- Standard Pipetting
- It is recommended for aqueous solutions
such as buffers, diluted acids or alkaline.
- This technique is commonly used when
pipetting a mixing sample or reagents
into another liquid.
(FIRST STOP ------- RELEASE TUMB ------PRESS
AGAIN THE FIRST STOP ---- SECOND STOP ------
RELEASE)
STEPS:
1. First, to aspirate the liquid in the tip, press the
plunger to the first stop (there are 2 stops)
immerse the pipette tip vertically in the liquid
(up)
2. Slowly release the plunger while the tip is
immerse, the liquid will be aspirated into the
pipette tip (up)
3. To dispense the liquid, place the tip on the
inner wall in receiving vessel in the tip angel
4. Slowly press the plunger to the first stop to
dispense the liquid
- For solutions with high viscosity or
tendency to foam used the reverse
techniques.
- This technique is suitable for dispensing
reagents or solutions that has a high
viscosity or tendency to fall easily.
- It is recommended for dispensing very
small volume.
(SECOND STOP ----- RELEASE TUMB ------ FIRST
STOP ------ RELEASE)
STEPS:
1. First, to aspirate the liquid in the tip, press the
plunger into the second stop and immerse the
pipette tip vertically in the liquid.
2. Slowly release the plunger while the tip is
immerse, the liquid will be aspirated into the
pipette tip.
3. To dispense the liquid, place the tip on the
inner wall of the tube at a straight angle.
4. Slowly press the plunger to the first stop to
dispense the liquid
5. Wipe the tip on the inner wall while taking
the tip of the inner vessel
To aspirate deretcho sa second stop
Release the plunger
First stop the release
TAKE NOTE: In Reverse Pipetting, the Residual  The middle number refers to 1x
Liquid Remains in the Tip for the Reverse Pipette microliter
 The button refers to 0.1x of microliters

In inserting the tip, the tips are rack in plastic

backsets with covers when you receive a box it
will be sterile, please be careful when touching
box or tips not contaminate them. So the box
should be close when not in use to prevent
airborne contamination.
And It Does Not Belong To Dispense Volume
How to insert tips?
Micropipettes in the laboratory has 3 sizes 1. Select the correct size tips
each of which measures a different range of 2. Open the box without touching the tips
volumes with your hands
Nutri sizes are the following: 3. Insert the micropipettes shaft into the tip
1. P20 and press down firmly, this will attached
the tip to the shaft
-Range of volume 0.5 to 20
4. Remove the micropippetor with the tip
microliter
attached, closed the box without touching
2. P200
the tip with your hands
- 20 to 200 microliter
3. P1000
- 100 to 1000 microliter Care of the micropipettes:
1. Avoid Mouth Pipetting
- 2. When we give back the pipette,
This sizes are noted on the top of the plunger Always in a Vertical Position to Avoid
button Contamination
3. Always Disinfect the pipettes after
It is important to always select the correct pipette Use
by thinking about which one is suitable pipetting 4. Avoid Plunger Button To Set Volume
the desired volume. Na Lalagpas Sa Designated Volume
Example: Niya
o You where ask to aspirate 80 microliters 5. Avoid Mabagsak Ang Pipette
sample. So will choose the P20
3. PASTEUR PIPETTE
P1000
 The top number refers to the 1000x of
microliter.
 The middle number refers to 100x microliter
 The button refers to 10x of microliters
- Also known as droppers
P200 - This are laboratory liquids handling tools
 The top number refers 100x microliters or used to transfer small quantities of
 The middle number refers to 10x liquids.
microliter - Traditionally made from clean glass,
 The button refers to 1x of microliters although now a days we use the plastic
- There is no volume, so this is only used
P20 for transfer pipette.
- RUBBER BULB ASPIRATOR (Yung Kulay
 The top number refers 10x microliters Red) serves as a vacuum source for filling
reagents trough a pipette or Pasteur
 pipette and also help to control the flow 7.CERAMIC-RINGED SLIDE
of liquid from the dropping button.
- Specific for VDRL usage

4. DISPOSABLE DISPENSER

8. SIX CELL AGGLUTINATION SLIDE

- It provides simple means of storing and - It is used for Agglutination test
dispensing liquid and other materials in - Frequently used of initial confirmation of
laboratory. specific pathogen

9. Plastic Card Slide

- It is used to determine the titer.

MICROTITER PLATE

- It is a flat plate with multiple wells used as a

small test tubes.

- consist of 96 wells (A-H and 1-12)

-convenient high through tools for organizing

5. VENIPUNCTURE SET
tissue culture or test such as HN Screening:
immunoassay (e. g. ELISA, radioimmunoassay and
fluorescent immunoassay.

WASH BOTTLE, STIRRING ROD, TUBERCULIN

SYRINGE WITH BUNT TIP NEEDLE,
LABORATORY FILM

6. KLINE SLIDE Wash bottle- it is a squeeze bottle used to wash

or rinse various pieces of glassware or plastic
- Usually made up of a clear glass and has ware in the laboratory.
depression to avoid mixing one’s cell to
another Stirring rod- it is used to stir or mix solution
- Use in agglutination test and used in
Tuberculin Syringe with Bunt tip Needle- it is used
widedal test and for the quantitative test,
to transfer reagent for VDRL Testing
VDRL test
- It handled with care, do not touched or Laboratory Film (para film)
wipe the test slide surface especially the
inner green area - used for covering or sealing vessels.
HENRY INSTRUMENT Parts:

Water Bath Eyepiece- it is a set of lenses closes to the eye

- is a laboratory equipment that used yo incubate
samples at a constant temperature over a long
period of time.
- preferred hit source for heating flammable
chemicals instead of an open to prevent ignition.
Mechanical Rotator
Nosepiece-rotating the houses objective lenses.
Objectives- used to magnify the object being
observed.
- each objective is mark with each magnification
factor.

- it is used to blend or agitate sample within flask Color:

or tubes.
Red- scanning objective (4x) total: 40x
Clinical centrifuge
Yellow- low power obj. (10x) total: 100x
-used in various laboratories to separate fluids or
gases liquid base on density. Blue- high power obj. (40x) total: 400x

-calibrated every 3 months and clean weekly basis
Hot Air Sterilizer
- it is one method of effectively killing microbes of
all kinds specially bacteria, virus on heat
persistance material.
- disinfect
White- oil immersion objective (100x) total:
1000x
Stage clips - method clips that holds slide in place
Stage - platform where the slide is place
Iris Diaphragm - adjust the amount of light that
reaches the specimen

Elisa microplate Reader Condenser -focuses light from the illuminator

-for Elise test
Heating Block
-commonly used to heat samples of preservation
and reaction, Dna amplification and
electrophoresis.
Compound microscope
1609- GALILEO GALILEI- invented the compound
microscope
- it is an upright microscope that uses to sets of
lenses, to obtain a higher magnification.
-it is used to view magnified images of small
objects on a glass slide.
until to the specimen being viewed.
Arms - it connects the body tubes to the base of
the microscope
Force adjustment knob -it brings the specimen
into general focus
Find adj. Knob -it finds tunes the focus and
increases the details of the specimen
Base - support the microscope
Body tube -it connects the eyepiece to the
objective lenses
Rheostat light control -regulates the intensity of
lights
Specimen on slide -specimen being examine
 PHLEBOTOMY
WHY DO YOU THINK IT IS NECESSARY TO
DRAW BLOOB?
-PHLEBOTOMIST play a very big part in
treating and diagnosing patients, if it wasn’t
for the laboratory testing, doctors would jus
be guessing, laboratory results are critical to
patient care.
Some of the most important aspects of the
laboratory test are the following:
1. DIAGNOSTING AND SCREENING
- to figure out what is wrong with the patient
2. THERAPEUTIC ASSESSMENT
- To develop the correct treatment or choose
the right drugs from the patients
3. MONITORING HEALTH
- to make sure that the therapy treatment is
working
- derived from the greek word:
PHLEB-veins/blood vessels
TOMY- to cut/make an incision
MOST COMMON METHOD OF TODAY
FOR BLOOD COLLECTING ARE:
1. VENIPUNCTURE
2. SKIN PUNCTURE
SKIN PUNCTURE
-Refered as dermal sticks, capillary dross,
finger sticks and heel stick
We draw blood from:
1. FINGERTIPS
2. EARLOBES
- LESS AT MIXTURE with tissue juice; less
pain, less free nerve ethics
3. LATERAL PORTION OF THE PLANTAR
SURFACE OF THE FOOT
- preferred for less than 1 year old children
also called HEEL
CAPILLARY PUNCTURE
- it refers to sampling blood from a puncture
on a finger or an earlobe
ADVANTAGES over venipuncture:
-it is less invasive
- it requires smaller amount of blood volume
-perform quickly and easily
Order of draw for capillary puncture:
1. BLOOD GASES
2. EDTA
3. OTHER ANTI COAGULANTS
4. SERUM TUBES
VENIPUNCTURE
- process of collecting or drawing blood from a
vein
- most common way to collect blood
specimens for laboratory testing
-most frequent procedure performed by a
phlebotomist and the most important step in
this procedure is PATIENT IDENTIFICATION
WHY DO WE PERFORM VENIPUNCTURE?
1. to obtain blood samples in order to perform
diagnostics
2. To monitor the levels of various blood
components
VENIPUNCTURE SITES
 ANTECUBITAL FOSSA
- preferred sites for blood collection
- it is where arm bends in the elbow
3 vein that are preferred for blood draw in the
location:
1. MEDIAN CUBITAL VEIN
- preferred site and most stable vein
2. CEPHALIC VEIN
- not preferred but can be used; it has the
tendency to roll
3. BASILIC VEIN
- closes to the heart; not recommended to
draw blood; it lies close to the brachial nerve
and artery
FRANCISCO
 PHLEBOTOMY
 TOURNIQUET
VENIPUNCTURE SETS: Standard size: 18x21 by 1 inch
 GLOVES -should be applied approximately 3-4 inches
- to protect us from sudden splash of blood; above the puncture sites (7.5-10cm)
avoid contamination
- it should not exide more than 1 min. In the
 NEEDLES arm of a patient
- used to collect blood sample; single used only
-if greater than 1 min. Has elapse, tourniquet
 THE HUB/ ADAPTER can be re applied after 2 mins.
- used with the evacuated collection tube
-if you are using bp cuff tourniquet- pressure
 ECT should be in 40-60mmHg
- they are designed to draw a pre-determine
volume of blood -prolong application of tourniquet can cause
- have different additives; used for collecting hemoconcentration
blood specimen
 EVACUATED TUBES
 ALCOHOL WIPES -order of draw in venipuncture:
- used for disinfect the side for extraction
HENRY’S 21ST ADDITION
 SYRINGES
- used to draw blood sample
Yellow sterile blood culture tube
 BANDAGES/TAPE/ GAUZE SPONGES citrate
- the one we put after we draw blood non-additive tube
heparin
 TOURNIQUET EDTA
- applied for detection of vein FLOURIDE Gray

 REQUISITION FORM  Syringed method

- request test by a physician -EDTA
-OTHER ANTICOAGULANT TUBES
 SHARP CONTAINER -NON-ADDICTIVE TUBE
- for disposal of needles SEROLOGY- RED TOP TUBE(mostly used)

 NEEDLE EDTA(LAVENDER)
- it is the anticoagulant of choice for
Two way needle: hematology cell counts and cell morphology.
Common length: 1-1.5 inches
Common gauge: 19, 20, 21(20-book) -preferred for platelet count; it is not used for
Angle of insertion: 15-30 degrees coagulation test
Color coded hub: mode of action: it chelates calcium
18- pink Inversions: 8
21-Green BLACK TOP
22-Gray - used for modified westergren ESR
23- /light blue Ratio: 1ml to 4ml of blood
25- orange
CITRATE
2 times attempt - for coagulation and platelets studies
Inversions: 3-4 times
Ratio:1x9

FRANCISCO
 PHLEBOTOMY

HEPARIN
-anticoagulant for osmotic fragility test
Inversions: 3-4 times

ALCOHOL
Antiseptic used: 70% isopropyl alcohol

-before drawing blood there should no traces

of alcohols that remain in the skin;allow it to
air dry

SOURCES OF ERROR IN VENIPUNCTURE

Errors in Venipuncture Preparation:

1. Improper patient identification

2. Failure to check patient adherence to dietary
restrictions
3. Failure to calm patient prior to blood collection
4. Use of improper equipment and supplies
5. Inappropriate method of collection

Errors in Venipuncture Procedure:

1. Failure to dry site completely after cleansing with

alcohol
2. Inserting the needle bevel side down
3. Use of needle that is too small
4. Venipuncture in an unacceptable area
5 Prolonged tourniquet application
6. Wrong order of tube draw
7. Failure to mix blood collected in additive tubes
immediately
8. Pulling back on syringe plunger too forcefully
9. Failure to release tourniquet prior to needle
withdrawal

Errors in Venipuncture Completion:

1. Failure to apply pressure immediately to

venipuncture site
2. Vigorous shaking of anti-coagulated blood
specimen
3. Forcing of blood through syringe needle into tube
4. Mislabeling of tubes
5. Failure to label appropriate specimens with
infectious disease precaution
6. Failure to put date, time, and initials on
requisition
7. Slow transport of specimens to laboratory

FRANCISCO