Manual Reticulocyte Count Procedure

Manual Reticulocyte Count Procedure 1. Add two drops of patient blood to three drops of prepared New Methylene Blue stain in a labeled tube. Mix well. 2. Incubate the mixture for 15 minutes at37∘C. 3. After incubation, resuspend the cells.

4. Make three thin peripheral blood smears from the resuspended mixture and let them dry. (Note: One slide will be used for testing. The other two slides will be used only if needed for troubleshooting).

5. Once the slides are dry, using the$$100x$$oll objective, observe the peripheral blood smear. 6. Count a total of 500 red blood cells (RBCs), identifying them each as either “mature red blood cells”, or “reticulocytes” as follows: a. Mature red blood cells: pale greenish-blue in color, no inclusions present inside the cells b. Reticulocytes: pale greenish-blue red blood cells, with deep blue staining inclusions inside the cell 7.

When counting, make sure to count in a systematic way, so that cells are not counted twice or missed. Do not count white blood cells, platelets, or other cellular debris. 8. Once 500 total RBCs have been counted, transcribe the number of reticulocytes identified to the reporting$$\log$$.

9. A different Medical Laboratory Scientist (MLS) must then repeat the 500-cell red blood cell count procedure on the same slide (steps 5-8). 10. If the counts from the two MLS do not match within$$+/-20\%$$, start over at step 5 using one of the alternate slides. If the two MLS counts match within$$+/-20\%$$, calculate the final percent reticulocyte count using this formula: % reticulocyte count =1000 RBCS  Total Reticulocyte Count ​×100Example Reporting$$\log \&$$Calculation Result interpretation The reference interval for an adult percent reticulocyte count is$$0.5-2.0\%$$. Increased percent reticulocyte counts may be seen in Sickle Cell Disease or Beta Thalassemia. Decreased percent reticulocyte counts may be seen in Aplastic Anemia or Anemia of Chronic Disease. Refer to the Mannal Reticulocyte Count Procedure above.

Question #3: A final percent reticulocyte count was reported as$$7.2\%$$. Assuming the procedure was performed correctly and from an adult female: Would her Reticulocyte Count be increased, decreased, or normal when compared to the reference interval? Would you believe that this result came from a patient with Aplastic Anemia? Why or why not?

Question #4: The following manual reticulocyte count was performed. If MLS #1 performed the procedure correctly, list two possible reasons why MLS #2’s reticulocyte count does not match within$$+/-20\%$$. Question #5: An MLS performing a manual reticulocyte count did everything correctly, except he/she incubated the blood/new methylene blue stain mixture for 5 minutes.

In this situation, would the final Percent Reticulocyte Count be unaffected, falsely decreased, or falsely increased? Explain.