Laboratory Activity 1 – Instrumentation in Serology Flashcards
coated
Solid Phase
coated
Solid Phase
uncoated
Liquid Phase
Containers of blood, serum, and plasma samples
Ordinary Test Tubes
Used to contain reactions (transparent)
Ordinary Test Tubes
For observing BOTH agglutination & hemolysis
Ordinary Test Tubes
(clotting of different cells)
agglutination
(test tube is used instead of glass slides)
hemolysis
For preparing red cell suspension (RCS)
Graduated Centrifuge Tube
solution containing rbc antigens
red cell suspension (RCS)
Isotonic solution with RBC antigens (will not cause alterations to red cells)
RED CELL SUSPENSION
Intact RBC
Isotonic solution with RBC antigens
Normal saline solution (NSS) concentration
0.85% to 0.90% NaCl in H2O
for optimum binding of antibody to antigen
2 – 5% concentration of NSS
Prepare a solution containing RBC only:
- Whole blood (RBC w/ Antigens and plasma proteins)
- NSS
Number of times NSS is used to wash out plasma proteins
(3x)
source of RBC antigen
RED CELLS
Used to dispense pre-determined volumes of solutions (large or small volumes)
Serological Pipette
Serological Pipette Parts
- Tip 2. Stem 3. Mouth
Steps: Serological Pipette
o Aspirate a volume of liquid.
o Enclose the mouth using the nondominant finger.
o Wipe the sides.
o Calibrate to zero.
o Dispense to a container.
DO NOT calibrate using the
dominant hand and thumb.
Controls the volume of solutions
Aspirators
Can dispense volumes in microliters
Micropipette
o Aspirate the 1st stop
o Dispense the 2nd stop
Forward pipetting
o Aspirate the 2nd stop.
o Dispense the 1st stop.
Reverse pipetting (for viscous solutions)
*plasma and serum – not viscous
Forward pipetting
Both Dispenses small volume of solutions
Single and Multiple Channel Automatic pipette
Can dispense solution per drops
Pasteur pipette
Disposable dispensers
With graduations (volumes indicated)
Should be calibrated
Disposable dispensers
Free-felling; vertical solutions
Droppers
accumulation of antibody = problematic concentration
Slanted Droppers
o To make sure that there are no excess antibodies introduced into the tubes
Free-felling; vertical
Found in test kits
Stirrer
Both for pipetting and stirring
Stirrer
Flat end and tubing
Stirrer
Can be used for aspirating small volumes of serum/plasma/blood
Tubing
o Used for mixing and stirring the contents
Flat end
has interaction with the needle portion of the pump
Nozzle
for puncturing
Tip
used for blood-sampling
Shaft and bevel
large samples; Blood donation and blood banking
- Blood collection
small samples
- Blood sampling
Open or closed system
Hypodermic syringe and needle
Blood sampling ONLY*
Open system ONLY
Blunt Tip Needle
o Not used for collecting multiple samples
o Single collection
Open system ONLY
No bevel that helps in the proper insertion of needle to the vein
Blunt Tip Needle
For dispensing appropriate or calibrated volume of liquids in drops
Blunt Tip Needle
Not used for blood collection
Blunt Tip Needle
Same w/ droppers: Free-felling; vertical solutions
Blunt Tip Needle
Used for collecting multiple samples
Evacuated tube system
to avoid contamination of or carry-over of samples by following order of draw
Closed system
A brand Large
Kline Agglutination Slide
indicates the max. level of correct mixture
Circles in KAS
Has Different colors – to provide contrast
Kline Agglutination Slide
– utilizes white bg
o Charcoal: black
o For better evaluation
Different colors – to provide contrast
For viewing and evaluating agglutination reactions w/ application of contrast
Kline Agglutination Slide
indicating presence or absence of agglutination
Qualitative
determining the titer (concentration of antibodies)
Titration
with depression – to avoid escape of mixtures; to avoid contamination of other wells
Ceramic-ringed slide
also used for viewing agglutination rx
Ceramic-ringed slide
– agglutination
o Slides
– agglutination & hemolysis
o Test tubes
o Microtiter plate –
agglutination
BOTH agglutination & hemolysis
Ordinary Test Tubes
agglutination ONLY
Kline Agglutination Slide
Ceramic-ringed slide
Microtiter plate
Six-cell Agglutination Slide
Plastic Card Slide
Microscope
disposable
Plastic Card Slide
1 container has 6 wells
Microtiter plate
Can contain small volumes of solutions to determine antibody concentration to look for other agglutination rx
Microtiter plate
1 aspiration only
Microtiter plate
Used by automated machines
Microtiter plate
– solid phase testing
- Open
– container only; no rgt/antibody/antigen attached
- Unopened/closed
Serologic rx – visible to naked eye
Microscope
Used for evaluating agglutination rx using LPO
Microscope
For checking possible microscopic rx
Microscope
For viewing microtiter plate
Inverted Microscope
For finding compatible organ donor
Inverted Microscope
o Top: light source
o Bottom: objectives
Inverted Microscope
Antibodies(ABs) – sensitive to temperatures
Water bath
o Reacts with enhanced temp
Antibodies(ABs)
↑temp
↑ rx
– warm-reacting at 37C (to enhance rx of ABs) in test tube
IgG
– does not react (cold AB)
IgM
Used for distinguishing cold vs. warm AB
Water bath
– interfere w/ testing results
Complement components/proteins
o Exposed to high temp = (loses chain and becomes non-functional)
HEAT DENATURATION
to heat inactivate proteins
o >55C = 56C
– to activate complement proteins
o Heat-inactivated serum
o Use of metallic plate
o Alternative: hot plate
Water contamination
Allows attraction of AB to AG
Mechanical rotator
Mechanical rotator Movement:
circular
Slides are used to enhance binding of AB to AG
Mechanical rotator
(as container of whole AB and AG can be utilized manually at exactly [?])
1 min
Application of force to allow binding/movement of AB to AG
A. Using slides
B. Using Test tubes
A. Mechanical rotator
B. Microtiter plate agitator
Application of titration to allow mixture of contents
Microtiter plate agitator
🤪
to enhance binding of AB to AG
A. Slides - 1
B. Test tube - 2
A. Mechanical rotator
B. Microtiter plate agitator
B. Clinical centrifuge
Enhances binding of AB to AG using test tubes
Clinical centrifuge
CC: for sample prep (plasma, serum)
IS: for enhancing agglutination rx
Clinical centrifuge
Test tubes as containers of samples/rgts for enhancing binding/movement of AB to AG
Clinical centrifuge
Used for preparation of RCS (washing)
Clinical centrifuge
Avoid contaminants
Hot-air sterilizer
Autoclave
Uses:
o For sterilizing equipment (heat under pressure)
o For treatment of metal plates
Autoclave