The FACSCalibur system is a modular, benchtop flow cytometer from Becton Dickinson Immunocytometry System (BDIS) that is easily modified based on research needs. It designed for applications that range from routine clinical to advanced research, analyzing single cells as they pass through a focused laser beam. It is a tricolor, automated system that is capable of analyzing a wide variety of reagents. The system offers many opportunities for personal optimization of aspects such as software control, auto-sample loading and push-button fluidic control, with maximum ease and productivity.
The FACSCalibur technical specification capabilities include a 488nm, air-cooled, Argon-ion laser, excitation/emission optics, fluidics and signal processing. There is a fluorescence sensitivity detection limit of 750 molecules equivalent soluble fluorescein per particle and a resolution coefficient of variation of less than 3% (full peak for propidium iodide-stained chicken erythrocyte nuclei). The optical capacity includes forward and side scatter sensitivity, enabling the separation of fixed platelets from noise, as well as, optimum resolution performance for resolving lymphocytes, monocytes and granulocytes. The sort modes include single cell exclusion and recovery. The data management system offers a remote diagnostics capability, enabling direct customer support-instrument interaction.
The FASCalibur Flow Cytometer is presently used in our lab for in vitro diagnostics, specifically, enumerating leucocyte (non-blast) subsets with the appropriate software. In addition, the FACSCalibur system is being used for many other research applications including multicolor analysis, retic counts, immunophenotyping and stem cell analysis.
TriTEST three-color direct immuno-florescence reagents were optimized to ensure consistent, reliable, direct enumeration of percent and absolute T-B and NK-lymphocyte subset counts in erythrocyte-lysed whole blood using the FACSCalibur. In one of our studies, we have used TriCD4/8/3 (BD) reagent and TriCD4/8/3 (Dinon A Ltd) reagent kits. TriTEST reagents were used with an isotype Immunotrol control (Beckman Coulter) to verify the accuracy of pipetting technique. The florochromes used were FITC, PE and PerCP. The data analysis and interpretation were carried out using the Cellquest software. This comparative analysis was carried out to check the efficacy of the two kits. The results indicated similar values with minimum variation between the kits, suggesting compatibility with the FACSCalibur system for both.
The BD FACSCalibur system with its user-friendly design, high sample throughput, improved workflow management and simple, yet sophisticated, software enhanced the quality of routine analysis in our laboratory. A strong research background combined with the assets of the BDFACSCalibur system virtually ensures successful flow cytometry in broad areas of biological and medical research. Fluorescence quantitation with BD Quantibrite reagents, platelet analysis, stem cell research, multi-parameter DNA analysis, BD cytometry bead array, fluorescent protein research and microbial analysis are just a few of the application possibilities using the BD FACSCalibur system.
Researchers and pathologists will appreciate the BD FACSCalibur system for the flexibility, which allows modification for various applications as research needs change. From molecular to cellular, this high-performance system is a valuable tool for assay, development, verification and physical separation of target cells for further analysis. In addition, the innovative reagents, powerful software and versatile hardware, meet the current demands of both clinical and research environments.
Dr. Mukesh Agrawal & Mr. Sharath Babu
Pathologist
Clinical Reference Lab
VIMTA Labs Ltd.