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Kaluza flow analysis software

Manufactured by Beckman Coulter
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Sourced in United States, France

Kaluza Flow Analysis Software is a data analysis and visualization tool designed for flow cytometry applications. It provides users with advanced features for managing, analyzing, and interpreting flow cytometry data.

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Lab products found in correlation

Gallios flow cytometer, by Beckman Coulter (21 mentions) Cytoflex flow cytometer, by Beckman Coulter (4 mentions) Propidium iodide, by Merck Group (4 mentions) Navios flow cytometer, by Beckman Coulter (4 mentions) Cytoflex, by Beckman Coulter (3 mentions) Rnase a, by Thermo Fisher Scientific (3 mentions) Facsverse flow cytometer, by BD (3 mentions) Brefeldin a, by Merck Group (2 mentions) Anti cd45 percp cy5, by Thermo Fisher Scientific (2 mentions) Cd279 pd 1, by BD (2 mentions) 7 aminoactinomycin d 7 aad, by Beckman Coulter (2 mentions) Annexin 5 fitc kit, by Miltenyi Biotec (2 mentions) Cyan adp, by Beckman Coulter (2 mentions) Anti mouse cd16 cd32 fc block, by BD (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Phosphate buffered saline (pbs), by Merck Group (2 mentions) Facscalibur, by BD (2 mentions) Mwreg30, by BioLegend (1 mentions) Gallios analyzer, by Beckman Coulter (1 mentions) Bovine lactadherin fitc, by Prolytix (1 mentions)

Close spelling variants of this product

Kaluza Flow Cytometry Analysis Software version 2.1 Kaluza flow cytometry analysis software version Kaluza flow cytometry analysis software version 1.2 Kaluza flow cytometry data analysis software Kaluza Flow Cytometry Analysis Software v1.1 Kaluza 1.2 flow analysis software Kaluza® 1.3 Flow Analysis Software Kaluza flow cytometry analysis software 1.2 Kaluza Flow Cytometry Analysis Software Kaluza Analysis Software

69 protocols using kaluza flow analysis software

1

Platelet Activation Profiling by Flow Cytometry

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Assessment of platelet activation by flow cytometry was performed by diluting washed platelets (1 × 106 platelets/ml) in Tyrodes buffer containing 1 mM CaCl2. Murine platelets were activated with thrombin (0.1 IU/ml) in the presence of anti‐mouse CD41‐BV421 antibody (Biolegend, Clone # MWReg30; 1:50), anti αIIbβ3 in active conformation (Emfret; clone JON/A‐PE; 1:25), P‐selectin‐APC (Biolegend, clone APM‐1; 1:25), or bovine Lactadherin‐FITC (Haematologic Technologies; 10 µg/ml). The activation was quenched at 5 min using ice‐cold 1% PFA Tyrodes buffer. Samples were run in the Gallios analyzer (Beckman Coulter). Studies were performed with n = 3–5 mice/day per group and repeated at least twice. Flow cytometry data were analyzed using Kaluza flow analysis software (Beckman Coulter) and Flowjo (Flowjo, LLC). Gating strategy as previously described (Davizon‐Castillo et al., 2019).
Davizon‐Castillo P., Allawzi A., Sorrells M., Fisher S., Baltrunaite K., Neeves K., Nozik‐Grayck E., DiPaola J, & Delaney C. (2020). Platelet activation in experimental murine neonatal pulmonary hypertension. Physiological Reports, 8(5), e14386.
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2

Comprehensive Immune Cell Profiling by Flow Cytometry

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Peripheral blood mononuclear cells (PBMC) were isolated by density centrifugation with Lymphoprep (Axis-Shield). PBMC or whole blood samples were stained with the following fluorochrome-conjugated monoclonal antibodies: CD3-efluor605, CD4-efluor450, CD27-APC-efluor780, HLA-DR-efluor780, CD45RA-efluor605, FOXP3-PE (eBioscience), CD4-APC-H7, CD8-Percp, CD8-PE-Cy7, CD31-AF647, CD45RO-FITC, CD45RO-PE-Cy7, CCR7-PE-Cy7, Ki-67-Percp-cy5.5, CTLA-4-BV421 (BD Biosciences), PD-1-PE, CD28-AF700 (Biolegend), and CD161-PE (Miltenyi Biotec). Intracellular staining for FOXP3, Helios, Ki-67, and CTLA-4 was performed after cells were permeabilized with a FOXP3 staining buffer set according to instructions of the manufacturer (eBioscience). Whole blood samples were treated with BD lysing solution according to the instructions of the manufacturer (BD Biosciences). Stained samples were analyzed on a LSR-II flow cytometer (BD Biosciences). Analysis was performed with Kaluza Flow Analysis Software (Beckman Coulter). Absolute numbers of CD3+ T cells, CD4+ T cells, CD8+ T cells, B cells, and NK cells were determined according to the MultiTest TruCount method (BD Biosciences), as described by the manufacturer. TruCount samples were measured on a FACSCanto-II (BD Biosciences) and analyzed with FACSCanto Clinical Software (BD Biosciences).
van den Brom R.R., van der Geest K.S., Brouwer E., Hospers G.A, & Boots A.M. (2018). Enhanced expression of PD-1 and other activation markers by CD4+ T cells of young but not old patients with metastatic melanoma. Cancer Immunology, Immunotherapy, 67(6), 925-933.
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3

Flow Cytometric Analysis of Cell Cycle in Macrophages

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The cell cycle of MΦ was studied by flow cytometry with the DNA-prep reagents kit according to manufacturer's instructions (Beckman Coulter SAS, France). Propidium iodide staining was analyzed by the GalliosTM flow cytometer (Beckman Coulter SAS, France) with 20 000 events per determination. The analysis and determination of the cell distribution in each phase of the cell cycle was achieved based on the Kaluza® Flow Analysis software (Beckman Coulter SAS, France).
Leblond M.M., Pérès E.A., Helaine C., Gérault A.N., Moulin D., Anfray C., Divoux D., Petit E., Bernaudin M, & Valable S. (2017). M2 macrophages are more resistant than M1 macrophages following radiation therapy in the context of glioblastoma. Oncotarget, 8(42), 72597-72612.
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4

Identification of Angiogenic Monocytes

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Spleen tissue-derived or PB MNCs were stained with PeCy7-anti-CD14, FITC-anti-CD16, APC-anti-Tie2, PE-anti-CD62L and PerCP-anti-CCR2 (R&D Systems, Minneapolis, MN or eBioscience, San Diego, CA). According to Venneri et al. [13 (link)] angiogenic monocytes, acquired by a flow cytometer (Navios™; Beckman Coulter, Inc, Brea, CA) and analyzed by Kaluza® flow analysis software (Beckman Coulter), were identified as Tie2+ CD14lowCD16brightCD62L−CCR2− cells. The appropriate isotype controls were used to set the markers for positive/negative cells (see Fig 1 and S1 Fig). We evaluated the percentage of total CD14+ cells in PB or spleen tissue-derived MNCs of patients with PMF and CTRLs; the percentage of Tie2+ cells was investigated on MNCs, CD14+, CD14lowCD16bright, and CD14lowCD16brightCD62L−CCR2− cells. Data were shown as median (range).
Campanelli R., Fois G., Catarsi P., Poletto V., Villani L., Erba B.G., Maddaluno L., Jemos B., Salmoiraghi S., Guglielmelli P., Abbonante V., Di Buduo C.A., Balduini A., Iurlo A., Barosi G., Rosti V, & Massa M. (2016). Tie2 Expressing Monocytes in the Spleen of Patients with Primary Myelofibrosis. PLoS ONE, 11(6), e0156990.
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5

Multiparametric T Cell Phenotyping

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The following antibodies were used in this study for T cell phenotyping: CD3-PerCP (clone SK7/Cat# 347344), CD25-APC AF700, CD4-Krome Orange (13B8.2/A96417), CD8-Pacific Blue (B9.11/A82791), CD3-APC (Beckman Coulter Inc. Brea, CA), Rat Anti-Mouse IgG1-APC (X56/550874) (BD Biosciences, San Jose, CA). PD-1-Percp Cy7 and TIM3-APC (BD Biosciences, San Jose, CA) were used as markers of T cell exhaustion. MUC1 antigen expression by tumor cells was measured using anti-MUC1, (Santa Cruz Biotechnology. Inc., Dallas, TX). CAR molecules were detected using Goat anti-human F(ab’)2 antibody conjugated with AlexaFluor647 (109–606-097) (Jackson ImmunoResearch Laboratories, Inc., West Grove, PA). Cells were stained with saturating amounts of antibody (~5uL) for 20 min at 4 °C, washed (PBS, Sigma-Alrich, St. Louis, MO), and then acquired on Gallios™ Flow Cytometer (Beckman Coulter Inc., Brea, CA). Analysis was performed using Kaluza® Flow Analysis Software (Beckman Coulter Inc.).
Bajgain P., Tawinwung S., D’Elia L., Sukumaran S., Watanabe N., Hoyos V., Lulla P., Brenner M.K., Leen A.M, & Vera J.F. (2018). CAR T cell therapy for breast cancer: harnessing the tumor milieu to drive T cell activation. Journal for Immunotherapy of Cancer, 6, 34.
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6

Monocyte IL-6 Expression Assay

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was measured in vitro as the capacity of monocytes to express IL-6, using the 1130h blood sample on each of the seven intensive recording days. Whole blood was stimulated with lipopolysaccharide (LPS) from Escherichia coli 0127-B8 (LPS 100pg/ml, Sigma-Aldrich), and then brefeldin A (10 ug/ml, Sigma-Aldrich) was added to the sample, which was incubated for 4 hours at 37 °C in a 5% CO2 atmosphere. Following fixation and permeabilization procedures (Intraprep™ Permeabilization reagents [Beckman Coulter]), fluorescence-conjugated antibodies were added (CD14 APC, CD45 KrO [both Beckman Coulter], IL-6 PE [BD Bioscience]) and samples incubated for 15 min at room temperature in the dark. Samples were vortexed, washed with phosphate-buffered saline solution (PBS 1X, Sigma Aldrich), and stored at 2–8 °C in the dark after re-suspension in 500 ul of PBS containing 0.5% formaldehyde. Preparations were analyzed within 24 hours using a Gallios™ flow cytometer (Beckmann-Coulter) at the Flow Cytometry Core at BIDMC, and 100,000 events were acquired. Percentage of IL6-positive monocytes (LPS-stimulated and spontaneous) were quantified using Kaluza® Flow Analysis software (Beckmann Coulter).
Simpson N.S., Diolombi M., Scott-Sutherland J., Yang H., Bhatt V., Gautam S., Mullington J, & Haack M. (2016). Repeating patterns of sleep restriction and recovery: Do we get used to it?. Brain, behavior, and immunity, 58, 142-151.
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7

Isolation and Analysis of Murine Mammary and Lung Cells

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Mammary glands and lungs were isolated from mice. Single cell suspensions of mammary glands were prepared as we previously described (Sharon et al., 2013 (link)). For dissociation of lung tissue, collagenase type-II was substituted with trypsin (Biological Industries; 03–051-5B). Cells were stained using the following antibodies: anti-PDGFRα-PE (eBioscience; 12–1401-81), anti-EpCAM-APC (Miltany Biotec; 130–102-234), anti-CD45-PerCP-Cy5.5 (eBioscience; 45–0451-82), anti-CD45-PE-Cy7 (eBioscience; 25–0451-82), anti-Annexin V-Alexafluor 488 (Invitrogen; A13201), anti-CD34–FITC (eBioscience; 11–0341-82), and anti-αSMA-FITC (Sigma; F3777). For intracellular staining, cells were fixed and permeabilized with BD Cytofix/Cytoperm Plus kit according to the manufacturer's protocol. DAPI was used to exclude dead cells (Molecular Probes; D3571). Analysis or sorting was done with BD FACSAria II, BD FACSAria Fusion or CytoFLEX Flow Cytometer (Beckman Coulter, Inc.). Data analysis was done with BD FACSDiva software (BD Biosciences), CytExpert 2.0 (Beckman Coulter, Inc.) or the Kaluza Flow Analysis software (Beckman Coulter, Inc.).
Raz Y., Cohen N., Shani O., Bell R.E., Novitskiy S.V., Abramovitz L., Levy C., Milyavsky M., Leider-Trejo L., Moses H.L., Grisaru D, & Erez N. (2018). Bone marrow–derived fibroblasts are a functionally distinct stromal cell population in breast cancer. The Journal of Experimental Medicine, 215(12), 3075-3093.
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8

Multicolor Flow Cytometry Analysis of Immune Cell Subpopulations

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Whole blood samples of the patients were collected and the fresh whole blood was analyzed by multicolor flow cytometry according to our previously published and optimized IPT protocols.17 18 (link) IPT was performed within 3 hours after the collection of whole blood. Data acquisition was performed on a Gallios Flow Cytometer (Beckman Coulter) in the standard filter configuration. The Kaluza Flow Analysis Software (Beckman Coulter) was used for data analysis. The immune cell subpopulations analyzed are visualized and specified in online supplemental sFigure 1.
Zhou J.G., Donaubauer A.J., Frey B., Becker I., Rutzner S., Eckstein M., Sun R., Ma H., Schubert P., Schweizer C., Fietkau R., Deutsch E., Gaipl U, & Hecht M. (2021). Prospective development and validation of a liquid immune profile-based signature (LIPS) to predict response of patients with recurrent/metastatic cancer to immune checkpoint inhibitors. Journal for Immunotherapy of Cancer, 9(2), e001845.
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9

Quantifying GFP Fluorescence in P. sonchi

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In order to quantify green fluorescent protein (GFPUV) mean fluorescence intensities (MFI) of P. sonchi SBR5 pBV2mp-gfpUV strains harboring CRISPRi plasmids and proper controls, cell cultures were analyzed by fluorescence activated cell scanning (FACS). Therefore, P. sonchi cells were cultured until reaching exponential phase and centrifuged for 10 min at 4000 rpm (5810R centrifuge, Eppendorf, Hamburg, Germany). The pellets were washed two times in NaCl 0.89%, and the OD of the cultures was adjusted to 0.5. The fluorescence of the cell suspension was measured using flow cytometer (Beckman Coulter, Brea, USA) and the data analyzed in the Beckman Coulter Kaluza® Flow Analysis Software. As described previously, the settings for the emission signal and filters within the flow cytometer for detection of GFPUV fluorescence were based on a 550/525 bandpass FL9 filter (Brito et al. 2017b (link)).
Brito L.F., Schultenkämper K., Passaglia L.M, & Wendisch V.F. (2020). CRISPR interference-based gene repression in the plant growth promoter Paenibacillus sonchi genomovar Riograndensis SBR5. Applied Microbiology and Biotechnology, 104(11), 5095-5106.
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10

Multicolor Flow Cytometry for Circulating ICs

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Whole blood samples of the patients were collected and analyzed by multicolor flow cytometry according to our previously published modularly immunophenotyping (IPT) protocols.11 12 (link) Direct antibody staining of whole blood samples was performed without previous isolation of blood mononuclear cells, which allows the detection of all types of circulating ICs, including granulocytes. IPT was performed within 24 hours after the collection of whole blood. Data were acquired on a Gallios Flow Cytometer (Beckman Coulter, Brea, California, USA) in the standard filter configuration. The Kaluza Flow Analysis Software (Beckman Coulter) was used for data analyses.
Hecht M., Gostian A.O., Eckstein M., Rutzner S., von der Grün J., Illmer T., Hautmann M.G., Klautke G., Laban S., Brunner T., Hinke A., Becker I., Frey B., Semrau S., Geppert C.I., Hartmann A., Balermpas P., Budach W., Gaipl U.S., Iro H, & Fietkau R. (2020). Safety and efficacy of single cycle induction treatment with cisplatin/docetaxel/ durvalumab/tremelimumab in locally advanced HNSCC: first results of CheckRad-CD8. Journal for Immunotherapy of Cancer, 8(2), e001378.
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