Results for cell types (%) obtained by the Guava and the conventional analyzers

Results for cell types (%) obtained by the Guava and the conventional analyzers. comparison with analysis using two conventional flow cytometers. Methods 26 dogs with lymphoproliferative disease (23 with lymphoma, 3 with lymphocytic leukaemia) were studied along with 15 controls (2 non-lymphoma lymph nodes and 13 non-leukemic bloods. Lymphocytes were immunostained with fluorescent-labeled, monoclonal antibodies against CD3 and CD21. To assess the effectiveness of the personal flow cytometer in discrimination between T and B cell immunophenotypes, T and B cell counts for half the samples (14 blood and 11 lymph node) were also determined using the same method and conventional flow cytometers (FACSCalibur, Cyan Dako). To assess the effectiveness of the personal flow cytometer in discriminating between leukocyte types, lymphocyte differential counts were determined for 21 blood samples and compared with those from automated hematology analyzers (CELL-DYN 3500, n=11 Turanose and ADVIA 2120, n=10). Quality and sub-cellular distribution of immunostaining was assessed using fluorescence microscopy. Results The protocol for immunophenotyping took 2 to 3 3?hours to complete from the point of receipt of sample to reporting of immunophenotype. The personal flow cytometer differential lymphocyte counts correlated highly (n=20; r=0.97, p 0.0001) with those of automated Turanose haematology analyzers. The personal flow cytometer counts consistently, but mildly, underestimated the percentages of lymphocytes in the samples (mean bias of -5.3%.). The personal flow cytometer immunophenotype counts were indistinguishable from those of conventional flow cytometers for both peripheral blood samples (n=13; r=0.95; p 0.0001; bias of -1.1%) and lymph node aspirates (n=11,r=0.98; p 0.001; bias of 1%). All but one leukemic and one lymphomatous lymph node sample, out of 26 samples of dogs with lymphoproliferative disease analyzed, could be immunophenotyped as either B or T cells. Conclusions We conclude that use of only 2 monoclonal antibodies is sufficient for immunophenotyping most cases of canine lymphoma by flow cytometry and Turanose enables rapid immunophenotyping. The personal flow cytometer may be as effectively used for immunophenotyping canine lymphoma as conventional flow cytometers. However, the personal flow cytometer is more accessible and user-friendly, and requires lower sample volumes. strong class=”kwd-title” Keywords: Immunophenotyping, Canine lymphoma, Personal flow cytometer, Microfluidics, Guava Background Lymphoma is one of the most prevalent cancers in dogs [1]. Diagnostic testing and prognosis is based on clinical signs and degree of spread, morphological features of the lymph node and lymphocytes, and other cytopathologic features such as mitotic rate, and clonality of antigen-receptor rearrangement or of cluster of differentiation (CD) antigens. Immunophenotyping CD antigens has contributed significantly to both diagnosis and prognosis of lymphoid neoplasia. This approach measures the binding of labelled, monoclonal antibodies to specific intracellular or surface CD antigens. It is well-established and has long been used in cell analysis, particularly in the fields of haematology and immunology [2-4]. For lymphoma, it can be accomplished using either immunohistochemistry of tissue-biopsy sections [5] or by immunocytochemistry of fine needle aspirates. Cytologic analysis can be done manually on smears using microscopy [6] or on cell suspensions using automated, flow cytometry. Immunophenotyping is most easily and rapidly accomplished by flow cytometry. Flow cytometry of blood, lymph node and bone marrow samples may improve evaluation and prognosis of dogs with lymphoma [4,7-9]. However, in veterinary medicine this technique is mainly available only as a research tool, rather than for widespread diagnostic use as occurs in human medicine [9]. There are only a few European laboratories that routinely provide immunophenotyping by flow cytometry for veterinary patients. The main barriers associated with the expansion of flow cytometry in veterinary medicine are the substantial cost of the analyser, reagents, and facilities, and the need for advanced training of the instrument operators. In addition, interpretation and analysis of results requires understanding and understanding of stream cytometry and its own concepts. It really is well Turanose noted that immunophenotype of neoplastic lymphocytes correlates considerably with the success time of canines with lymphoma and it is of significant worth in prognosis [10-15]. In 175 canines with lymphomas, T-cell phenotype acquired shorter relapse-free period (52 versus 160?times, p 0.001) and shorter success situations (153 versus 330?times, p 0.001) than B-cell phenotype [15]. Dobson, Blackwood et al. 2001 discovered that the T- cell phenotype is normally connected with a considerably shorter recurrence-free period and reduced success times. Hazard proportion for T-cell versus B-cell immunophenotype lymphoma was 4, with 95% self-confidence period from 1.4 to 11.3, p=0.035. Nevertheless, the relationship of immunophenotype with prognosis isn’t perfect, and exclusions have been discovered. For instance, the very clear and small T-cell lymphoma provides one of the Rabbit Polyclonal to MRPL2 better prognoses [11]. For prognosis Thus, immunophenotyping data can be used and also other cytological and clinical assessments of amount of.