Immunology and Serology Laboratory Manual

Group No: 2A

ACTIVITY NO. 1 Date of Performance: September 12-14, 2021

Submission Deadline:
Date: September 15,
INSTRUMENTATION IN 2021
SEROLOGY Time: 12:00 PM

Instructor: MS. PADUA, DAWN KIMBERLY

g: CONTENT: NEATNESS:
ni
at THIS PORTION WILL BE FILLED BY THE
r ________ ________ RECEIVING END:
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Gr Date of Submission: ______________________
OVERALL:
Time of Submission: ______________________

Received by: ____________________________

GROUP MEMBERS CONTRIBUTIONS

LEADER: ESPIRITU, Rustie V. Compilation, 1 (c, d)
MEMEBERS:
CATBAGAN, Andrei Eleazar R. 1 (a, b)
MACALANDA, Reden R. 1e (a, b)
BRABANTE, Ivy Lloren D. 1e (c, d)
BURCENA, Jewel Anne D. 1 (f, g, h)
GAERLAN, Mari Bianca R. 1 (i)
PADUA, Kyla Leeanne V. QFR 1
PIMENTEL, Yves Laure QFR 2
QUITILEN, Dawn Princess C. QFR 3
SIAZAR, Jamby Mae S. QFR 4

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 Immunology and Serology Laboratory Manual
Activity No. 1
INSTRUMENTATION IN SEROLOGY
WRITE-UP SHEET

I. Attach and label the image, describe and give the uses of the following instruments and
equipment.
A. Graduated Centrifuge tube Description and Use/s:

Also called centrifuge tips, are made out of either

glass of plastic which resembles test tubes with
tapered tips. Used in centrifugation, a test that is
utilized to separate the components of a specific
sample by rapid rotation along its fixed axis.

B. Pasteur pipette Description and Use/s:

Bulb draw A disposable, single use pipette made out of thin

glass or plastic with a tapered tip. Mainly used in
handling solutions individually such as medium
transfer and handling. It is also capable of moving
small amounts of liquid without needing precise
measurement.

C. Serological Pipettes Description and Use/s:

It is frequently used in the laboratory for transferring

milliliter volumes of liquid, from less than 1 ml to up
to 50 ml. It has Etched band in the upper part, a top
graduation mark, and a calibrated tip. It is also
essential in mixing cell suspensions.

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 Immunology and Serology Laboratory Manual
D. Automatic Pipette Description and Use/s:

It can be manually held yet provides a highly

accurate instrument capable of precise handling of
very small liquid samples. It is also used in the
biological, biochemical, and microbiological
teaching and research laboratories. These are
adjustable pipette.

E. Reaction Slides:
a. Kline Slide Description and Use/s:

Usually made up of a clear glass and has

depressions to avoid mixing of one cell to another.
It is used in agglutination test and used in widal test
and for quantitative VDRL test to detect syphilis
caused by Treponema pallidum subsp. Pallidum.
Detection of Typhoid fever caused by Salmonella
typhi

b. Six Cell Agglutination Slide Description and Use/s:

To check white agglutination Detection of Anti-

Streptolysin O in Streptococcal infxns. Also for C-
reactive Protein an indicator of inflammation.

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 Immunology and Serology Laboratory Manual
c. Plastic Card Slide Description and Use/s:

Plastic card slide or plastic-coated disposable card

is used in Rapid Plasma Reagin (RPR) test which is a
modified VDRL test involving macroscopic
agglutination. The procedure includes placing a
patient serum (approximately 0.05 mL) in an 18-mm
circle on a plastic-coated disposable card using a
capillary tube or Dispenstir device. A positive test
result may mean that the patient has syphilis.

d.Ceramic Ringed Slide Description and Use/s:

The Ceramic Ringed slide is used in VDRL slide

qualitative test on serum with the test principle:
heat-inactivated serum is pipetted into one ring of
a ceramic ringed slide and is mixed with a buffered
saline suspension of cardiolipin–lecithin–
cholesterol antigen and this mixture are
microscopically examined for flocculation. The
ceramic ringed slide is used because glass slides
with concavities, wells, or glass rings are not
recommended for this test.

F. Microtiter Plate Description and Use/s:

Microtiter plates are high-throughput organizing

tools for tissue culture, PCR testing (such as HIV
screening), and immunological assays including
ELISA, RIA, and FIA. Other uses for PCR plates
include enzyme reaction analysis and chemical
identification using colorimetric or spectroscopic
methods.

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 Immunology and Serology Laboratory Manual
G. Mechanical Rotator Description and Use/s:

Mechanical Rotator are used to agitate or mix

materials in flasks or tubes. These devices are made
comprised of a housing that houses the motor and
control panels, as well as an agitation platform.

H. Water Bath Description and Use/s:

A water bath is a piece of laboratory equipment that

keeps samples at a steady temperature for an
extended length of time. To avoid ignition, a water
bath is preferable over an open flame while heating
combustible substances.

I. Equipment for ELISA

a. Heating Block Description and Use/s:

A heat block, otherwise known as a dry bath

incubator, is a microprocessor-controlled heating
block with heater technology. A heat block is most
commonly used to heat samples of preservation and
reaction, DNA amplification, and electrophoresis.
With its digital precision and convenience, the
instrument is suitable for various applications that
require accurate results and temperature control.

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 Immunology and Serology Laboratory Manual
b. Microstrip Reader Descriptioon and Use/s:

Microstrip reader is a laboratory instrument intended

for in-vitro diagnostic use. It is a compact,
microprocessor-controlled, photometer system
designed to read and calculate the results of endpoint
colorimetric assays, which are read in microtiter
strips.

c. Microplate Washer Description and Use/s:

Microplate washers tremendously improve the speed

and accuracy of many different washing procedures,
and are particularly useful for Enzyme-Linked
Immunosorbent Assays (ELISAs). Microplate washers
are also employed to wash cell cultures, protein
arrays, Western blots and beads as well as applied in
DNA purification protocols.

II. QUESTIONS FOR RESEARCH

1. What is the importance of instrumentation in Serology laboratory?

ANSWER: Maintenance should be performed and monitored by the staffs who are trained to use the equipment
and instruments in the laboratory. Important function, testing, and an easy and fast diagnosis are significant.
Serology Laboratory is where we study fluids in the blood especially antibodies, with the support of laboratory
instruments. Instruments used for measurements including dilutors, spectrophotometers, spectrometers, and
automatic pipettes to hold a specified amount of liquid and are calibrated to transfer or deliver that exact amount
as marked on their surfaces. Special handling and testing should be met and observe. Instruments and
equipment should be monitored and check routinely for the accuracy and precision when the equipment is tested
under working conditions.

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 Immunology and Serology Laboratory Manual

2. How should serological pipettes and automatic micropipette be calibrated?

ANSWERS: A frosted ring at the no calibrated end, with calibrations continuing to the tip, distinguishes the
serologic pipette. The initials TD (to deliver) are imprinted on the pipette, and each size has a color-coded ring
that specifies the volume for fast identification. Normally, the serologic pipette is allowed to drain naturally. To
provide the full volume, the remaining drop must be evacuated, depending on the calibration. Serological
pipettes should be calibrated all the way to the tip and the final drop blown out. Serological or 'blow-out' pipettes
are calibrated to deliver the complete volume of the pipette by blowing the last drop of liquid out of the tip. (You
may not need to blow these out if you are not aiming to deliver the entire volume of the pipette.)

Automatic pipettes provide for quick, repeatable measuring and administration of equal-volume solutions. The
piston-operated plunger in the sampling kind of automatic pipette is mechanically operated. These can be
adjusted to deliver different volumes of reagent or sample with the same device. Before using automatic pipettes
and micropipettes, they must be calibrated. To eliminate any residual liquid, pre-rinse the tip by aspirating and
dispensing the indicated volume three times. Aspirate the calibration volume without forming bubbles and
slowly pour the liquid into the weigh boat. Then, on the balance, record the weight and repeat the process ten
times.

3. Discuss the principle of Enzyme-linked Immunosorbent Assay (ELISA).

ANSWER: The enzyme-linked immunosorbent assay (ELISA) is a method for capturing target antigens (or
antibodies) in samples using a specific antibody (or antigen) and detecting/quantifying target molecules using
an enzyme reaction with its substrate.

In an ELISA test, different antigen-antibody combinations are employed, always with an enzyme-labeled antigen
or antibody, and enzyme activity is calorimetrically assessed.

The enzyme activity is determined by changing the color of a substrate when it is modified by the enzyme. The
product's light absorption following the addition of the substrate is measured and translated to numeric values.
The assay is referred to as a direct ELISA, indirect ELISA, sandwich ELISA, competitive ELISA, and so on,
depending on the antigen-antibody combination.

4. What are the advantages of using flow cytometric analysis instead of ELISA?

ANSWER: The potential benefits of automated flow cytometric methods include the opportunity to obtain
outcome in short time frames, monitoring the frequency of MN and the DNA distribution of MN concurrently, as
well as using flow sorting to further examine MN using other methodologies. The real advantage is that they
significantly decrease the time expense linked with isolation task. Elisa, on the other hand, is based on antigen-
antibody reactions wherein people may very well be poor producers of antibodies or have some intruding
substance in their blood. As an outcome, the portion of antibody may be too low to precisely measure or may go
undiagnosed, resulting in limited blockage of immobilized antigen and inaccurate results.

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 Immunology and Serology Laboratory Manual
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