Phagocytosis from peritoneal and blood leukocytes was acquired on an AMNIS Flowsight flow cytometer (Merck-Millipore, Darmstadt, Germany) interfaced with a DELL computer. Data from 20,000 events were obtained, utilizing the 488 nm laser, at a 20x magnification, using the INSPIRE software. Direct measurements of mean fluorescence in the green channel were recorded as phagocytosis. Phagocytosis percentage (PP) was recorded as the number of cells engulfing at least one zymosan particle (with green fluorescence divided by the total number of cells [multiplied by 100]). Acquired data were analyzed using IDEAS analysis software (EMD Millipore) version 6.1 for windows.
Ideas analysis software
Manufactured by Merck Group
The IDEAS analysis software is a robust and versatile tool designed to facilitate the analysis of data generated from various experimental techniques. It provides a user-friendly interface and advanced algorithms to enable efficient and accurate data processing and interpretation. The core function of the IDEAS software is to offer a comprehensive platform for researchers to explore and derive insights from their experimental findings.
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Lab products found in correlation
2 protocols using ideas analysis software
1
Quantifying Phagocytosis in Leukocytes
2
Evaluation of Antigen Presentation in moDCs
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MoDCs (5 × 105) were seeded in a 48-well plate and stimulated for 24 h with 3 μg/mL Sa lysate or 100 ng/mL LPS with or without PSMα3 (10 μM) or PSMα3 alone prior to the incubation with Ovalbumin (OVA)-AlexaFluor647 (5 μg/mL, Invitrogen) together with PSMα2 FITC (0.5 μM) for 30 min at 37°C, 5% CO2. Unspecific binding of OVA/PSMα2 was assessed by incubating the cells on ice. Cells were washed twice with ice-cold PBS containing 2% FBS. Subsequently, cells were blocked and stained with ZombieAqua (BioLegend), HLA-DR BV650 (L243, BioLegend), HLA-DR APC-Cy7 (L243, BioLegend) and CD11c PE-Cy7 (Bu15, BioLegend) as described above and analyzed by flow cytometry or by multispectral imaging flow cytometry. For the latter, images of 10,000 living moDCs were acquired using the Image-Stream mkII (Amnis) with the INSPIRE instrument controller software. The data were analyzed using the IDEAS analysis software (Merck Millipore).
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