title: Comparison of white blood cell differential percentages determined by the in-house LaserCyte hematology analyzer and a manual method.
abstract: BACKGROUND
  The LaserCyte hematology analyzer (IDEXX Laboratories, Chalfont St. Peter, Bucks, UK) is the first in-house laser-based single channel flow cytometer designed specifically for veterinary practice. The instrument provides a full hematologic analysis including a 5-part WBC differential (LC-diff%). We are unaware of published studies comparing LC-diff% results to those determined by other methods used in practice.
  
  
  OBJECTIVE
  To compare LC-diff% results to those obtained by a manual differential cell count (M-diff%).
  
  
  METHODS
  Eighty-six venous blood samples from 44 dogs and 42 cats were collected into EDTA tubes at the Forest Veterinary Centre (Epping, UK). Samples were analyzed using the LaserCyte within 1 hour of collection. Unstained blood smears were then posted to Langford Veterinary Diagnostics, University of Bristol, and stained with modified Wright's stain. One hundred-cell manual differential counts were performed by 2 technicians and the mean percentage was calculated for each cell type. Data (LC-diff% vs M-diff%) were analyzed using Wilcoxon signed rank tests, Deming regression, and Bland-Altman difference plots.
  
  
  RESULTS
  Significant differences between methods were found for neutrophil and monocyte percentages in samples from dogs and cats and for eosinophil percentage in samples from cats. Correlations (r) (canine/feline) were .55/.72 for neutrophils, .76/.69 for lymphocytes, .05/.29 for monocytes and .60/.82 for eosinophils. Agreement between LC-diff% and Mdiff% results was poor in samples from both species. Bland-Altman plots revealed outliers in samples with atypical WBCs (1 cat), leukocytosis (2 dogs, 9 cats), and leukopenia (16 dogs, 11 cats). The LaserCyte generated error flags in 28 of 86 (32.6%) samples, included 7 with leukopenia, 8 with lymphopenia, 7 with leukocytosis, 1 with anemia, and 1 with erythrocytosis. When results from these 28 samples were excluded, correlations from the remaining nonflagged results (canine/feline) were .63/.65 for neutrophils, .67/.65 for lymphocytes, .11/.33 for monocytes, and .63/.82 for eosinophils.
  
  
  CONCLUSIONS
  Although use of a 100-cell (vs 200-cell) M-diff% may be a limitation of our study, good correlation between WBC differentials obtained using the LaserCyte and the manual method was achieved only for feline eosinophils.
authors.affiliation: Langford Veterinary Diagnostics, School of Clinical Veterinary Science, University of Bristol, Langford, Bristol, UK. kos.papasouliotis@bristol.ac.uk
authors.forename: Kostas
authors.initials: K
authors.lastname: Papasouliotis
authors.forename: Simon
authors.initials: S
authors.lastname: Cue
authors.forename: Elsa
authors.initials: E
authors.lastname: Crawford
authors.forename: Mark
authors.initials: M
authors.lastname: Pinches
authors.forename: Michel
authors.initials: M
authors.lastname: Dumont
authors.forename: Ken
authors.initials: K
authors.lastname: Burley
mesh_terms: D000818:Animals; D001322:Autoanalysis / Q000662:veterinary; D002371:Cat Diseases / Q000097:blood / Q000175:diagnosis; D002415:Cats / Q000097:blood*; D004283:Dog Diseases / Q000097:blood / Q000175:diagnosis; D004285:Dogs / Q000097:blood*; D004804:Eosinophils; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods / Q000592:standards / Q000662:veterinary*; D007958:Leukocyte Count / Q000295:instrumentation / Q000379:methods / Q000592:standards / Q000662:veterinary*; D018655:Lymphocyte Count / Q000295:instrumentation / Q000379:methods / Q000592:standards / Q000662:veterinary; D009504:Neutrophils; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric; D014730:Veterinary Medicine / Q000295:instrumentation* / Q000379:methods / Q000592:standards