Centrifuge Calibration

CALIBRATION OF SEROLOGIC CENTRIFUGES
PURPOSE
Each centrifuge should be calibrated upon receipt and after adjustments or repairs. The
centrifuges should be calibrated at least yearly, and when the quarterly rpm reading
exceeds the 200 rpm range determined during the yearly calibration. Calibrating the
centrifuge evaluates the mechanics of the centrifuge (behavior of cells in solutions of
different viscosity); it does not test the reactivity of different antibodies.
PROTOCOL
1. Serum: Dilute anti-D 1:1000 to give at least a 1+ to 2+ macroscopic reaction.
2. Control cells: Screening Cells I or II for the Rh positive control cell.
A1 or B Reverse Cells for the Rh negative control cell.
3. Prepare a 12 X 75mm tube for each positive control and a duplicate set of tubes for
each negative control.
Add the serum and control cells to each tube just before centrifugation.
4. In pairs, one positive and one negative, centrifuge the tubes for the designated times
on the log sheet.
(ex. 20 seconds, 25 seconds and 30 seconds) Observe each tube for agglutination
and record
observations as indicated.
5. The optimum time of centrifugation is the least time required to fulfill these criteria:
a. Agglutination is as strong as determined in preparing reagents.
b. The red cell button is clearly delineated and the periphery is sharply defined, not
fuzzy.
c. The supernatant fluid is clear.
d. The red cell button is easily resuspended.
6. Washing procedure: Manually wash tubes with saline. In pairs, one positive and one
negative, centrifuge
tubes for the different times. All red cells should be in a clearly delineated button
with few cells remaining
up the side of the tube. The least time required to accomplish this is the optimum
washing time for the
centrifuge.
If >30 seconds is required for either spinning or washing, repeat the procedure with
new time.
Record in "other" area on log.
CENTRIFUGE CALIBRATION PROCEDURE
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
1
TUBE LABEL D+ cells (I or II) D neg cells (A1 or B) Anti D (1:000)
1 (neg 20) 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.
(Example on next page)
2
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.
REFERENCES
United Blood Services, Fargo, ND
AABB Technical Manual, 12th Edition, 1996. Pages 715-716.
3
4-29-92 br revised 6/95, 1/98 12/00 br
_____________________________________________________
Medical Director Review/Date
AVERA ST. LUKES BLOOD BANK

CENTRIFUGE CALIBRATION LOG
CENTRIFUGE ID NUMBER __________________________ DATE/INITIALS ________________
CELLS 20 SEC 25 SEC 30 SEC OTHER

Supernatant
Clear
(YES or NO)
A. IS
POS B. 37C
C.
Wash
D. AHG
A. IS
NEG B. 37C
C.
Wash
D. AHG
Button
Delineated
(YES or NO)
A. IS
POS B. 37C
C.
Wash
D. AHG
A. IS
NEG B. 37C
C.
Wash
D. AHG
Resuspend Easil
(YES or NO)
A. IS
POS B. 37C
C.
Wash
A. IS
NEG B. 37C
C.
Wash
Strength of
Reaction
A. IS
POS B. 37C
D. AHG
A. IS
4
NEG B. 37C
D. AHG
Interpretation: SPIN Time ________________ RPM________________

WASH Time_______________ RPM Range________________
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CALIBRATION OF SEROLOGIC CENTRIFUGES

CENTRIFUGE CALIBRATION PROCEDURE

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)

AVERA ST. LUKES BLOOD BANK

CENTRIFUGE ID NUMBER __________ DATE/INITIALS

CELLS 20 SEC 25 SEC 30 SEC OTHER

Interpretation: SPIN Time RPM

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