[MLAB 1415] Laboratory
Specimen through Differential
Laboratory: Specimen through Differential
Skills= 20
Objectives:
1. To appropriately run a specimen on the Micros 60.
2. To prepare and stain a blood smear using the automated stainer,
3. To determine the relative number of each type of white cell present in the blood by
performing differential cell counts on normal and abnormal blood smears.
4. To determine within one qualitative unit the red cell, white cell, and platelet
morphology of each of the blood smears.
5. To determine within ± 30% accuracy an estimate of the white cell counts and the
platelet counts of each of the blood smears.
Principle:
This laboratory will combine all the skills learned previously as well as enable the
student to practice using a hematology analyzer and automated stainer.
Specimen:
Peripheral blood smear made from EDTA-anticoagulated blood. Smears should be
made within 4 hours of blood collection from EDTA specimens stored at room
temperature to avoid distortion of cell morphology. Unstained smears can be stored
for indefinite periods in a dry environment, but stained smears gradually fade
unless coverslipped.
Reagents, supplies, and equipment:
1.
2.
3.
4.
5.
6.
7.
Patient sample provided by instructor or sample collected in class
Micros 60 instrument
Automated Stainer
Manual cell counter designed for differential counts
Microscope
Immersion oil
Lens paper
[MLAB 1415] Laboratory
Specimen through Differential
Procedure:
Prepare Slide
1. Invert the sample gently 8-10 times and look for clots.
2. Prepare two (2) smears according to the procedure outlined in the Blood Smear
Preparation and Stain Lab and allow them to dry.
Run Sample
3. Before running the first sample of the day, run the Start Up by pressing the
Start up key and following the prompts on the LED read out. Verify that the
Background checks are all “0”. If they are, continue. If there is a background
value that is not “0”, run another start up cycle. If there is a value other than
“0” after the second start up, consult with the instructor.
4. On the Micros 60, manually enter the patient name by pressing the ID key. Use
the up and down arrows to enter alphabetical characters. Press <Enter> after
each alphabetical entry.
5. After the patient ID is input, press <Enter>
6. Gently and thoroughly mix the patient sample.
7. Unstopper or uncap the specimen and immerse the aspirator tip in the sample.
8. Press and release the sample bar (behind the aspirator needle)
9. Remove the sample tube when the instrument beeps.
10. Review the data obtained from the printer and answer questions 1-5 on the
report form prior to performing the manual differential.
Stain Slide
11. Stain the dried slides by placing them with the colored/frosted end towards you
with the written patient information facing to the left in the groove on the
conveyor spirals on the right side of the stainer as shown below.
[MLAB 1415] Laboratory
Specimen through Differential
Perform Manual Differential
12. Focus the microscope and scan the smear on the 10X objective (low power).
13. If the smear is acceptable, estimate the white cell count by counting the
number of WBCs in ten (10) 50X fields.
14. Using the 100 X oil objective, examine the smear for platelet morphology and
perform a PLT estimate in five (5) 100X fields.
15. Perform a 100 or 200 WBC differential as indicated by the automated WBC
count.
16. Begin the count in the thin area of the slide and utilize the battlement track to
read the slide.
17. Count each white cell and nucleated RBC seen and record on a differential cell
counter, until 100 (or 200) white cells have been counted. Record your results
on the data sheet.
18. Examine the red cell morphology in a thin area of the slide. Note any variations
from normal and classify them according to the grading system given in the RBC
Morphology Lab.
19. All abnormal morphology (RBC, WBC, or PLT) is reported in the Morphology
section of the report form for each patient/slide. If you do not observe
abnormal morphology, write “normal” in this section.
Procedural Notes
1.
When in the clinical world, before reporting significant abnormalities such as
blasts, malaria or other significant finding on a patient’s differential, ask a more
experienced tech to review the smear for confirmation. In clinical settings
where a pathologist or hematologist is present, the smear is set aside for
Pathologist Review. Review the WBC & PLT Estimates/Inclusions Lab and
RBC Morphology Lab for ACC operating procedure for reporting peripheral
smear findings.
2.
When the WBC is very high (>40,000/μL), a 200-cell diff should be performed
to increase the accuracy of the diff. The results are then divided by 2 and a
note made on the report that 200 white cells were counted.
REFERENCES
1. Rodak, Bernadette, Hematology—Clinical Principles and Applications, 4th edition, pp202-204.
2. Turgeon, Mary Louise, Clinical Hematology - Theories and Procedures, 3rd edition, pp318-319.
3. Seton Medical Center Hematology Procedure Manual
4. Mckenzie, Shirlyn, Clinical Laboratory Hematology,2nd edition, pp.772-773.
MLAB 1415: Hematology
Specimen through Differential
Report Form
Skills: 20 pts.
Student Name:__________________________________
Date:_____________________________
From the patient’s automated CBC printout, answer the questions 1-5.
1.
2.
3.
4.
5.
Patient Name
and ID or
Slide Number
Do you expect to observe anisocytosis on the peripheral blood smear? _________
Do you expect to observe hypochromia? _______
Is the WBC count within the adult reference range? ______
Does the patient have relative Lymphocytosis? ______
Is the patient microcytic, normocytic, or macrocytic? _______________
WBC Estimate
PLT Estimate
Neutrophil %
Pro:
Atyp:
Myelo:
Meta:
Band:
Lymph
Seg:
Total:
Total:
Instructor use only:
Lymphocyte
%
Slide labeled correctly ___
Slide acceptable ___
Specimen run correctly___
Monocyte
%
Eosinophil
%
Basophil %
Morphology
Others
%
# of
nRBCs/100
WBCs