1. An example of Flow Cytometric DNA Analysis as a diagnostic tool J. Chezar Western Galilee Hospital-Nahariay Israel

2. History: A 49 year old male referred to the hematology department because of pancytopenia. After 2 dry taps, a bone marrow aspirate resulting in barely 200 uL of material was obtained and sent for flow cytometry. The sample was stained with the following antibody combinations. CD45PC7/CD34ECD/CD33 PC5/ CD117 PE/ HLA-DR FITC, CD45PC7/CD3ECD/CD56PC5/CD8PE/CD4FITC+CD19FITC, CD45PC5/CD13PE/CD16FITC, CD45PC5/CD64PE/CD14FITC. DNA cell cycle analysis was performed alone and in combination with CD45 FITC and glycophorin A FITC, CD13 FITC. In addition a bone marrow biopsy was sent to pathology. Immuno- histochemistry was done with peroxidase conjugated LCA, MPO, CD34, TdT, CD15, CD117, PAS, CD43, CD138, CD31, CD79a, Factor VIII, Ker, CD68, CD3, B and T cell antibodies

3. BLASTS

4. Normal Bone Marrow Patient Bone Marrow Aberrant population

5. Myeloid phenotype = Blasts?

6. MPO: GRAN+ / BLASTS -

7. PAS NEG IN BLASTS

8. Normal 2n Aneuploid (A)

9. To verify identity of aberrant population – double stain with PI for DNA and relevant FITC labeled antibody

10. A Aneuploid population NOT leukocytic

11. A Aneuploid population NOT thrombocytic

12. A Aneuploid population IS erythroid !! Glycophorin A +

13. Glycophorin

14. Erythro Leukemia DNA analysis and subsequent double staining with appropriate antibodies lead to the correct diagnosis.