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Kaluza 1.2 flow analysis software

Manufactured by Beckman Coulter
Sourced in United States

The Kaluza 1.2 flow analysis software is a tool designed for the analysis and visualization of flow cytometry data. It provides a user-friendly interface for data management, gating, and data analysis. The software supports standard flow cytometry file formats and offers a range of data display options, including histograms, dot plots, and contour plots.

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5 protocols using kaluza 1.2 flow analysis software

1

Flow Cytometry Analysis of T Cell Subsets

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The following monoclonal antibodies were used: anti-CD3-e450 (OKT3), anti-CD8a-APC-e780 (OKT8) (eBioscience, Vienna, Austria), anti-CD45RO-FITC (UCHL1), anti-CCR7-PE (3D12) (BD Bioscience, Breda, Netherlands), and anti-CD28 PeCy7 (CD28.2) (Biolegend, Uithoorn, The Netherlands). Cells were sorted using a MoFlo flow cytometry cell sorter (Backman Coulter, Woerden, The Netherlands).
Expression of cell surface markers on T cells was assessed using mAbs against human CD28-PE-CY7 (CD28.2) (Biolegend), CCR7-PE (3D12), and CD45RO-FITC (UCHL1) (BD Biosciences). Cells were analyzed using a BD LSR-II Flow Cytometer and the Diva software (BD Biosciences). Data analysis was done using the Kaluza Flow Analysis Software (1.2) (Beckman Coulter).
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2

T Cell Surface Marker Analysis

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Cell surface markers on in vitro activated T cells were assessed using mAbs against human CD25-PE (BC96) (eBioscience, Vienna, Austria) and CD45RO-FITC (UCHL1) (BD Biosciences, Breda, The Netherlands). Cells were analyzed on BD LSR-II Flow Cytometer by Diva software (BD Biosciences). Data analysis was performed on Kaluza Flow Analysis Software (1.2) (Beckman Coulter, Woerden, The Netherlands).
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3

Comprehensive B Cell Immunophenotyping

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Blood was collected in EDTA tubes, and 100 μl was incubated with anti-human CD19-eFluor450 (eBioscience, San Diego, CA, USA), CD24-fluorescein isothiocyanate (FITC) (BD biosciences, Franklin Lakes, NJ, USA), CD27-APC-eFluor780 (eBioscience), CD38-PeCy5 (eBioscience), CD5-PerCp-Cy5.5 (Biolegend, San Diego, CA, USA) or the corresponding isotype controls. After 15 minutes cells were treated with fluorescence-activated cell sorting (FACS) Lysing solution (BD Biosciences). Samples were measured using an LSR-II flow cytometer (BD biosciences) and data were analyzed using Kaluza 1.2 flow analysis software (Beckman Coulter, Brea, CA, USA). B cells were divided into transitional, memory, naive, CD24highCD38high and CD24highCD27+ B cells as described previously [14 (link)]. CD5+ B cells were gated on an isotype control.
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4

Flow Cytometric Analysis of CD4+ T-cells

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The data were analyzed using Kaluza 1.2 flow analysis software (Beckman Coulter).
As stimulation reduces surface expression of CD4 on T-cells, CD4+ T-cells were identified indirectly by gating on CD3-positive and CD8-negative lymphocytes [15 (link)]. The unstimulated samples were used as a guide for setting the linear gates to delineate positive and negative populations for the cytokine staining. For surface staining, isotype controls served as controls.
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5

B cell subset analysis in GPA

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EDTA blood samples from GPA patients and HCs were washed twice with PBS + 1% BSA to remove plasma proteins. Next, 100 µl of the cell suspension was stained with CD19-eFluor450, CD27-APC-eFluor780 (both eBioscience, San Diego, CA, USA), IgD-PE (BD Biosciences, Franklin Lakes, NJ, USA), or the corresponding isotype controls. After 15 min, cells were fixed and erythrocytes were lysed using FACS lysing solution. Samples were washed and measured using an LSR-II flow cytometer (BD Biosciences) and data were analyzed using Kaluza 1.2 flow analysis software (Beckman Coulter, Brea, CA, USA). B cells were divided based on their surface expression of IgD and CD27. Results are expressed as percentages of total CD19+ B cells.
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