EXPERIMENT 1: QFR

#1 Glasswares:
a. Plain test tubes:
 15x100mm:container for sample
 13x100mm:container for sample
 12x75mm: as reaction tubes
 8x75mm:as reaction tubes
b. Graduated centrifuge
- for RCS preparation
-are used in laboratory centrifuges, machines that spin samples in order to separate solids out of
liquid chemical solutions. The centrifuge tubes can be made of glass or plastic, and resemble
miniature test tubes with tapered tips.
c. Pasteur pipet
-used to transfer blood samples from tubes and vials.
-commonly referred to as Transfer Pipets, are an essential product widely used by hospital,
research and industrial laboratories for transferring liquids and clinical samples
d. Serologic pipets
-is frequently used in the laboratory for transferring milliliter volumes of liquid, from less than 1
ml to up to 50 ml. The pipettes can be sterile, plastic, and disposable or sterilizable, glass and
reusable. Both kinds of pipettes use a pipet-aid, for the aspiration and dispensation of liquids.
Different sizes of pipettes can be used with the same pipet-aid for a variety of experimental
assays.
- for measurement and transfer
e. Glass syringe
-are preferred for specialty applications where plastic syringes are not suitable, such as for
locating the epidural space during the loss of resistance method for regional anesthesia.
Borosilicate glass syringes are far less likely to interact with fluids than plastic, and are often
chosen when biocompatibility may be a concern. They are also ideal when a very smooth action
is needed for specific applications, such as in regional anesthesia.
f. Plain glass slide
- is a thin flat piece of glass, typically 75 by 26 mm (3 by 1 inches) and about 1 mm thick, used to
hold objects for examination under a microscope. Typically the object is placed or secured
("mounted") on the slide, and then both are inserted together in the microscope for viewing.
- for blood typing
g. Blood typing slide
- are specifically designed for blood testing and will increase accuracy when blood typing.
 OTHER MATERIALS USES IN IMMUNOHEMATOLOGY

1. Blood Bag  Storage and preservation of the blood for upto 35 days.
Blood are used in blood transfusions (hemotherapy)
 Use to collect Plasma, RBC, WBC, and platelets
2. Tourniquet  constricting or compressing device used to control
arterial and venous blood flow to a portion of an
extremity for a period of time
 used to palpate veins for blood collection
3. Applicator stick  used for routine spot test sampling and drop transfers, or
wherever a handy, disposable mixing aid is required
4. Test tube Rack  Used to hold upright multiple test tubes at the same
time. It is most commonly used when various different
solutions are needed to work with simultaneously, for
safety reasons, for safe storage of test tubes, and to ease
the transport of multiple tubes.
5. Rubber pipettol  Serves as a vacuum source for filling reagents through a
pipette or pasteur pipette and also help control the flow
of liquid from the dropping bottle.
6. Wash Bottle  Squeeze bottle with a nozzle, used to rinse various pieces
of laboratory glassware, such as test tubes and round
bottom flasks. Wash bottles are sealed with a screw-top
lid.
7. Lancet  Similar to a small scalpel but with a double-edged blade
or needle. Lancets are used to make punctures, such as a
fingerstick, to obtain small blood specimens. Blood
lancets are generally disposable. Lancets are also used to
prick the skin in skin testing for Bleeding time.
8. Cotton Balls in petri  shallow cylindrical glass or plastic lidded dish that
dish biologists use to culture cells
9. Labelling Tape  has a special adhesive ability
 used to label tubes during testings
10. Parafilm or Nescofilm  semi-transparent, flexible, thermoplastic film with two-
way stretch
 used to seals tubes to avoid contamination
 2.b. How is the calibration done in a clinical centrifuge and water bath?

CLINICAL CENTRIFUGE CALIBRATION PROCEDURE:

Serum: Dilute anti-D 1:1000 to give at least a 1+ to 2+ macroscopic reaction.
Control cells: Screening Cells I or II for the Rh positive control cell.
A1 or B Reverse Cells for the Rh negative control cell.
1. Add the following to each of the appropriately labeled tubes.
TUBE LABEL D+ cells (I or II) D neg cells (A1 or B) Anti D (1:000)
1 (pos 20) 1 drop 2 drops
2 (pos 25) 1 drop 2 drops
3 (pos 30) 1 drop 2 drops

TUBE LABEL D+ cells (I or II) D neg cells (A1 or B) Anti D (1:000)

1 (neg 20) 1 drop 2 drops
2 (neg 25) 1 drop 2 drops
3 (neg 35) 1 drop 2 drops

Spin in pairs 1 pos/1 neg for the times noted on the log sheets. Record the observations on the sheets.
2. Add 2 drops of OAES, mix and incubate for 15 minutes. After incubation, work with only one set
of tubes at a time. Leave the other tubes in the dri-bath.
3. After incubation, mix and spin each set of tubes according to the times noted on each set of tubes.
Record observations.
4. Wash all tubes with saline (manually) three times according to the times noted on each set of
tubes. Record observations. Answer only questions 1 and 2 for this step.
5. Add 2 drops of AHG to each tube. Spin according to the times noted on each set of tubes.
6. Determine the optimal time for "spin time" and "wash time" according to the results obtained
from the above procedures. Indicate the times on a piece of tape and put the tape on the centrifuge
cover. Date/initial.
7. Perform the RPM of each centrifuge after the spin and wash times have been determined. Record
on the log sheet. This will be utilized each quarter for maintenance rpms.

WATER BATH CALIBRATION PROCEDURE:

1. Fill water bath and place thermometer into the bath. Cover the bath with foil to prevent
external temperature fluctuation.
2. After temperature has reached desired point – read the temperature on the water bath and
thermometer. If temperature accuracy is within desired tolerance skip steps 3 and 4.
3. Turn off water bath and turn back on while pressing “set” on the front panel of the water bath.
Adjust calibration by increasing or decreasing the temperature calibration value to match the
thermometer read.
4. Turn off the water bath to restart.