Xn 1000 as 01 automated hematology analyzer

Sample film preparations was developed using a volume of 100 µL of 10⁵ PMN/mL suspension, spread-out on glasses by cito-spinning centrifugation, fixed with methanol 10% for 10 min and stained with Giemsa. After washes with water, glasses were dried at room temperature overnight. Glasses were observed at 40× magnification and photographed using an Olympus IX-81 microscope equipped with a DP71 camera (Olympus, Tokyo, Japan).
Additionally, the transformation of poly-lobed morphology of normal PMN to mono-nuclear (MN) cells can indicate the kinetics of viability loss secondary to the incubation of PMN with CBD detected as the PMN/MN cells ratio, by Sysmex XN-1000 AS-01 Automated Hematology Analyzer. So, the PMN/MN rate can reflex the nuclear condensation index related to unviable PMN amounts.

Total and differential cells account of viable PMNs in suspension were developed using the Sysmex XN-1000 AS-01 Automated Hematology Analyzer (XN-Series, Sysmex^® Chuo-ku, Kobe, Hyogo 651-0073, Japan), in a mode that allowed for distinguishing the mono-nuclear (MN) cells from neutrophils. These counts were confirmed manually with Neubauer´s chamber, using a 1/400 dilution of the PMNs suspension in Turk’s solution (1–2% acetic acid with aqueous methylene blue).
The PMNs’ viability was measured with equal volume of Trypan Blue solution 0.4% using a 1/100 dilution of the abovementioned neutrophils suspension. PMN/MN ratio as well as the cell´s viability was determined before and after each experiment, and only cells suspensions with viability >95% were used.