Flow cytometric evaluation of canine bone marrow based on intracytoplasmic complexity and CD45 expression.
BACKGROUND
Because of the complexity, subjectivity, time, and technical skill required for determination of manual bone marrow differential cell counts, an alternative method is needed. Several flow cytometric methods have been described, but all have limitations.


OBJECTIVE
The purpose of this study was to evaluate a technique for bone marrow differential cell counting based on flow cytometric evaluation of CD45 expression and intracellular complexity (CD45 scatter plots).


METHODS
Bone marrow was obtained from 15 dogs that were being evaluated for hematologic disorders. In preliminary studies, the location of bone marrow subpopulations in the CD45 scatter plots was evaluated by labeling bone marrow with lineage-specific markers. A template was developed to identify these cell populations. Gates were set to identify granulocytes, myeloblasts, monocyte/macrophages, lymphocytes, and nucleated erythroid populations.


RESULTS
The CD45 labeling technique accurately quantified granulocytes, myeloblasts, erythroid precursors, and lymphocytes in canine bone marrow. Correlation coefficients with manual counts for granulocytes, myeloblasts, erythroid cells, lymphocytes, and monocyte/macrophages were 0.90, 0.89, 0.96, 0.91, and 0.54, respectively.


CONCLUSIONS
The capacity of the CD45 scatter-plot technique to quantify lymphocytes and myeloblasts is an advantage over previously described techniques. The simplicity of the CD45 labeling method and the ease with which batches of samples can be analyzed makes the technique potentially applicable as a routine test in clinical and research laboratories.
Weiss
Douglas J
DJ
Department of Veterinary PathoBiology, College of Veterinary Medicine, University of Minnesota, St Paul, MN., USA. weiss005@maroon.tc.umn.edu
D000818:Animals; D001853:Bone Marrow; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D002452:Cell Count / Q000662:veterinary; D004283:Dog Diseases / Q000175:diagnosis* / Q000276:immunology; D004285:Dogs; D005434:Flow Cytometry / Q000379:methods / Q000662:veterinary*; D006402:Hematologic Diseases / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary*; D017493:Leukocyte Common Antigens / Q000032:analysis*
D017493:Leukocyte Common Antigens