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Establishing Conditions for Blood Smear Drying and Staining on Sysmex SP-50 for Leukocyte Differential Count
Background:
The purpose of this study was to determine the staining conditions and appropriate fan1 start time (FAN1ST) for Sysmex SP-50 to produce blood smears (BS) that reflect the true lymphocyte morphology of patient samples.
Methods:
Using different start times of fan1, we obtained a set of 84 blood smear slides from 21 blood samples and measured 10,920 lymphocyte areas, which were then converted to compare lymphocyte sizes. We also performed a leukocyte differential count using Sysmex DI-60 on 202 blood smear slides prepared before and after the change in staining conditions and compared the results.
Results:
The mean lymphocyte sizes at FAN1ST 0 second, 5 seconds, 10 seconds, and 30 seconds were 12.55 μm, 12.14 μm, 11.27 μm, and 10.50 μm, respectively. The mean differences in the preclassification of neutrophils, lymphocytes, monocytes, eosinophils, and basophils in DI-60, according to the SP-50 staining conditions, were 0.88, -1.58, -0.24, 0.37, and 0.07, respectively.
Conclusions:
Wright-Giemsa staining of blood smears prepared on the SP-50 showed that changing the pH of the concentrated phosphate buffer to 6.6 and adjusting the staining time did not affect the results of the leukocyte differential count. However, since fan1 was used to dry the blood smear on the SP-50 and the lymphocyte size gradually decreased as the start time was delayed, it was necessary to set a start time for fan1 that did not affect the lymphocyte size.
Insights
Optimizing the fan1 start time (FAN1ST) on the Sysmex SP-50 is crucial for accurate lymphocyte size measurement in blood smears. Delayed drying times reduce lymphocyte size, impacting diagnostic accuracy.
Area of Science:
- Hematology
- Diagnostic Cytology
- Laboratory Automation
Background:
- Accurate assessment of lymphocyte morphology is vital for diagnosing various hematological conditions.
- Automated blood smear preparation systems require precise parameter optimization for reliable results.
Purpose of the Study:
- To determine optimal staining conditions and fan1 start time (FAN1ST) for the Sysmex SP-50.
- To ensure blood smears accurately reflect true lymphocyte morphology in patient samples.
Main Methods:
- Blood smears were prepared using varying FAN1ST on the Sysmex SP-50 from 21 blood samples.
- 10,920 lymphocyte areas were measured to compare sizes.
- Leukocyte differential counts were performed using Sysmex DI-60 on slides prepared under different staining conditions.
Main Results:
- Mean lymphocyte size decreased with delayed FAN1ST (0s: 12.55 μm, 30s: 10.50 μm).
- Changes in SP-50 staining conditions minimally affected leukocyte differential counts by DI-60 (e.g., lymphocytes: -1.58%).
Conclusions:
- Wright-Giemsa staining optimization (pH 6.6, adjusted time) did not impact leukocyte differential counts.
- FAN1ST is critical as delayed drying reduces lymphocyte size, necessitating a setting that preserves accurate morphology.
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