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Kaluza flow cytometry software

Manufactured by Beckman Coulter
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Kaluza flow cytometry software is a powerful data analysis tool developed by Beckman Coulter. It is designed to provide comprehensive and intuitive analysis of data generated by flow cytometry instruments. The software enables users to visualize, analyze, and interpret complex flow cytometry data with ease.

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Lab products found in correlation

Gallios flow cytometer, by Beckman Coulter (4 mentions) Gallios 10 color flow cytometer, by Beckman Coulter (4 mentions) Cyto id autophagy detection kit, by Enzo Life Sciences (2 mentions) In situ cell death detection kit, by Roche (2 mentions) Nim dapi solution, by Beckman Coulter (2 mentions) Galliostm flow cytometer, by Beckman Coulter (2 mentions) Dihydrorhodamine 123 dhr123, by Merck Group (2 mentions) Pbs 1x, by Thermo Fisher Scientific (2 mentions) Dylight405 conjugated anti human igm, by Jackson ImmunoResearch (2 mentions) Pe conjugated anti human iga, by Southern Biotech (2 mentions) Cytoflex flow cytometer, by Beckman Coulter (2 mentions) Msc phenotyping kit, by Miltenyi Biotec (1 mentions) Alexa fluor 647 azide, by Thermo Fisher Scientific (1 mentions) Click it chemistry, by Thermo Fisher Scientific (1 mentions) Propidium iodide, by Merck Group (1 mentions) Tryple express, by Thermo Fisher Scientific (1 mentions) Fitc annexin 5 apoptosis detection kit 1, by BD (1 mentions) Gallios 10 3 flow cytometer, by Beckman Coulter (1 mentions) Prism v8, by GraphPad (1 mentions) Round bottom polystyrene tubes, by Corning (1 mentions)

24 protocols using kaluza flow cytometry software

1

Apoptosis and Autophagy Quantification

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Apoptosis induction and development of acidic vesicular organelles, were quantified using in situ Cell Death Detection Kit (Roche Applied Sciences, Germany) and Cyto-ID™ Autophagy Detection Kit (Enzo LifeSciences, Plymouth Meeting, PA) respectively and analyzed by flow cytometry. Cells were stained for 30 min at 37°C, harvested and green (510–530 nm) fluorescence emission from 1 × 104 cells illuminated with blue (488 nm) excitation light was measured with a fluorescent activated cell sorter (FACS). A total of 50000 events were collected and data analysis was performed by using Kaluza flow cytometry software (Beckman coulter, Fullerton, CA, USA).
Bhardwaj M., Paul S., Jakhar R., Khan I., Kang J.I., Kim H.M., Yun J.W., Lee S.J., Cho H.J., Lee H.G, & Kang S.C. (2017). Vitexin confers HSF-1 mediated autophagic cell death by activating JNK and ApoL1 in colorectal carcinoma cells. Oncotarget, 8(68), 112426-112441.
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2

Flow Cytometry Profiling of MSCs

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To determine MSC surface marker expression cells were detached by accutase treatment and stained with MSC phenotyping kit (Miltenyi Biotech GmbH, Bergisch Gladbach, Germany) according to manufacturer’s instructions. Stained cells (5 × 105 cells per aliquot) were resuspended in 300 µL flow cytometry buffer and acquisition was carried out on a Gallios flow cytometer (Beckman Coulter, Brea, CA, USA). Between 1–5 × 104 gated events were recorded. Subsequent analysis was performed with Kaluza Flow Cytometry software (version 1.3, Beckman Coulter, Brea, CA, USA).
Egger D., Schwedhelm I., Hansmann J, & Kasper C. (2017). Hypoxic Three-Dimensional Scaffold-Free Aggregate Cultivation of Mesenchymal Stem Cells in a Stirred Tank Reactor. Bioengineering, 4(2), 47.
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3

Quantifying Caspase-3 Activation in Apoptosis

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Caspase-3 activation was analyzed by using a NucView 488 caspase-3 substrate assay kit (Biotium, Hayward, CA). The principle of this assay consists of a fluorogenic substrate which is cleaved by an active caspase-3 releasing a product that after binding to the DNA emits a bright green fluorescence signal. HL-60 cells were plated in 24-well plates (100,000) and exposed to Pyr-1 for 6 h. Controls were included as previously described. After the incubation period, cells were harvested in flow cytometry tubes and centrifuged at 262g, for 5 min. Cell pellets were disrupted by adding 200 μL of PBS containing 5 μL of NucView 488 Caspase-3 substrate (5 μM final concentration), and cell suspensions were incubated at room temperature for 30 min in the dark. After this step, 300 μL of PBS was added to the cell mixture and immediately analyzed by flow cytometry. Cells emitting a green positive signal were denoted as apoptotic (caspase-3 active). Approximately 10,000 cells (events) were collected per sample and analyzed by using Kaluza Flow Cytometry Software (Beckman Coulter).
Gutierrez D.A., DeJesus R.E., Contreras L., Rodriguez-Palomares I.A., Villanueva P.J., Balderrama K.S., Monterroza L., Larragoity M., Varela-Ramirez A, & Aguilera R.J. (2019). A new pyridazinone exhibits potent cytotoxicity on human cancer cells via apoptosis and poly-ubiquitinated protein accumulation. Cell biology and toxicology, 35(6), 503-519.
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4

Flow Cytometry Analysis of Lymphocytes

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Cells were incubated with mAbs at RT for 30 min in the dark, then washed and collected in PBS for flow cytometry analysis. All flow cytometry measurements were performed using a Gallios 10-color-flow-cytometer equipped with Kaluza flow cytometry software (both Beckman Coulter). The acquisition and analysis gates were restricted to the lymphocyte gate based on characteristic properties of the cells in forward and side scatter. At least 105 cells were acquired for the analysis.
Jeske S.S., Schuler P.J., Doescher J., Theodoraki M.N., Laban S., Brunner C., Hoffmann T.K, & Wigand M.C. (2020). Age-related changes in T lymphocytes of patients with head and neck squamous cell carcinoma. Immunity & Ageing : I & A, 17, 3.
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5

Cell Cycle Analysis of Compound F18

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An asynchronous culture of CEM cells was seeded in 24-well plates at a density of 100,000 cells/well in 1 ml of complete culture media and treated with concentrations of compound F18 for 72 h. Controls included in this experiment were DMSO, hydrogen peroxide, and untreated cells, as described above. Cells were treated with 50 μM, 25 μM, or 10 μM of compound F18. Concentrations of experimental treatments were chosen based on the CC50 value of the given compound. After 72 h, cells were collected, centrifuged at 262 g for 5 min, and resuspended in 100 μL of complete culture media. Then, 200 μL of a nuclear isolation medium (NIM)-DAPI solution, which simultaneously permeabilizes the plasma membrane and stains DNA with the violet-excited DNA-intercalating agent DAPI, was added to each sample and immediately analyzed by flow cytometry. Approximately 100,000 events (cells) were analyzed per sample to obtain a well-defined cell cycle profile using the Kaluza flow cytometry software (Beckman Coulter) [76 (link)]. All experimental treatments and controls were assessed in triplicate.
Tilekar K., Upadhyay N., Hess J.D., Macias L.H., Mrowka P., Aguilera R.J., Meyer-Almes F.J., Iancu C.V., Choe J.Y, & Ramaa C.S. (2020). Structure guided design and synthesis of furyl thiazolidinedione derivatives as inhibitors of GLUT 1 and GLUT 4, and evaluation of their anti-leukemic potential. European journal of medicinal chemistry, 202, 112603.
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6

Apoptosis Analysis of MCF-7 Cells

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To measure apoptosis, MCF-7 cells were seeded at 2 × 105 cells/ml in 6-cm culture dishes, cultured for 1 day, and treated with 5, 10, or 20 μg/ml n-hexane fractions of KYC4048 extract for 72 h. Cells were then harvested, washed with ice-cold PBS, fixed at 4°C for 24 h with ice-cold 75% ethanol, and pelleted by centrifugation at 1,500 ×g for 5 min at 4°C. Subsequently, cells were washed with PBS, treated for 1 h with 0.5 μg/ml RNase A in PI buffer, and stained for 30 min in the dark with 20 μg/ml PI at 37°C. Cell cycle distribution was then analyzed on the basis of DNA content using Kaluza flow cytometry software (Beckman Coulter, USA). Apoptosis was also quantified by flow cytometry based on annexin V-FITC and PI double staining. Briefly, MCF-7 cells were exposed to n-hexane fractions of KYC4048 metabolites, trypsinized, collected by centrifugation, resuspended in annexin V-FITC binding buffer (500 μl), and incubated at room temperature for 30 min in the dark with 1 μg/ml annexin V-FITC and 10 μg/ml propidium iodide. Samples were then analyzed with the Kaluza flow cytometry software.
Park J., Yoo H.J., Yu A.R., Kim H.O., Park S.C., Jang Y.P., Lee C., Choe W., Kim S.S., Kang I, & Yoon K.S. (2021). Non-Polar Myxococcus fulvus KYC4048 Metabolites Exert Anti-Proliferative Effects via Inhibition of Wnt/β-Catenin Signaling in MCF-7 Breast Cancer Cells. Journal of Microbiology and Biotechnology, 31(4), 540-549.
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7

Flow Cytometric Analysis of Cell Proliferation

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Flow cytometry was performed as previously described38 (link). Briefly, cells were stained for EdU (Invitrogen, Cat. No. A10044) incorporation by linking an Alexa Fluor 647 Azide (Thermo Fisher Scientific, Cat. No. A10277) to the incorporated EdU using Click-it chemistry (Invitrogen, Cat. No. C-10420). Genomic DNA was stained with propidium iodide (Sigma, Cat. No. 81845) after treatment of the cells with DNase-free RNase (Roche, Cat. No. 11119915001). Data were acquired on a Gallios flow cytometer (Beckman Coulter) and analyzed by Kaluza flow cytometry software (Beckman Coulter).
Padayachy L., Ntallis S.G, & Halazonetis T.D. (2024). RECQL4 is not critical for firing of human DNA replication origins. Scientific Reports, 14, 7708.
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8

Annexin V Apoptosis Assay in JEG3 Cells

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JEG3 cells treated with CER+Q-VD-OPh were dissociated using 0.5 ml per well TrypLE express (Invitrogen), washed and processed in accordance with the FITC Annexin V Apoptosis Detection Kit 1 (BD Pharmingen, San Jose, CA, USA) at a concentration of 1 × 106 cells/ml. In brief, Annexin V and/or propidium iodide were added to the cells and incubated alongside a binding buffer for 15 min at room temperature protected from light, then thoroughly washed. Data were acquired using the Beckman Coulter Gallios 10/3 flow cytometer and analyzed with Kaluza flow cytometry software (Beckman Coulter, Brea, CA, USA). Annexin V-positive events were measured using the FL-1 channel (525 emission maximum), whereas the FL-4 channel (670 nm emission maximum) was used for propidium iodide-positive events. Unstained cells, cells stained only with FITC Annexin V and cells stained only with propidium iodide were included as controls.
Bailey L.J., Alahari S., Tagliaferro A., Post M, & Caniggia I. (2017). Augmented trophoblast cell death in preeclampsia can proceed via ceramide-mediated necroptosis. Cell Death & Disease, 8(2), e2590-.
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9

Flow Cytometry Immune Phenotyping from Whole Blood

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The procedures to perform blood immune phenotyping on fresh-whole-blood samples and flow cytometry antibodies and fluorochromes used are described in the online supplemental methods and table S1, respectively. Unsupervised analysis of flow cytometry data were performed using t-distributed stochastic neighbor embedding (t-SNE) algorithm with the online R software (V.3,5,0, cytofkit2 package). After setting the compensation matrix, CD19 +or CD3+cells events were extracted and software transformation was applied, t-SNE analysis was achieved on 2000 CD19 +for each sample, using ‘B-cell’ panel markers. Flow cytometry gating strategy were performed as previously described.31 (link) Supervised analysis of flow cytometry data were performed using Kaluza Flow Cytometry Software (Beckman Coulter) and was done by a single operator, blinded to the clinical patients’ information. Graphs for flow cytometry data were performed by GraphPad Prism V.8.0 (GraphPad Software) and by R software (V.4.1.1; packages corrplot).
Carril-Ajuria L., Desnoyer A., Meylan M., Dalban C., Naigeon M., Cassard L., Vano Y., Rioux-Leclercq N., Chouaib S., Beuselinck B., Chabaud S., Barros-Monteiro J., Bougoüin A., Lacroix G., Colina-Moreno I., Tantot F., Boselli L., De Oliveira C., Fridman W.H., Escudier B., Sautes-Fridman C., Albiges L, & Chaput-Gras N. (2022). Baseline circulating unswitched memory B cells and B-cell related soluble factors are associated with overall survival in patients with clear cell renal cell carcinoma treated with nivolumab within the NIVOREN GETUG-AFU 26 study. Journal for Immunotherapy of Cancer, 10(5), e004885.
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10

Cell Cycle Analysis of HL-60 Cells

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HL-60 cells were seeded in 24-well plates at a concentration of 100,000 cells containing 1 mL of culture media per well. Cells were treated with 0.25 μM (CC25) and 0.39 μM (CC50) of Pyr-1 for a 72-h period. The controls included for this series of experiments were as previously described. Cells were subsequently collected in flow cytometry tubes, centrifuged at 262g for 5 min, and gently re-suspended in 100 μL of fresh culture media. Then, 200 μL of nuclear isolation medium (NIM)-DAPI solution (NPE Systems, Inc. Pembroke Pines, FL, USA and Beckman Coulter) was added to the cell suspension and immediately analyzed via flow cytometry. The NIM-DAPI reagent is able to permeabilize and stain the nuclei of the cells allowing to quantify their DNA content by using an FL-9 detector and the 405-nm laser for excitation purposes (Gallios). This FL-9 detector captures the fluorescence signal emitted when DAPI is intercalated to DNA (461 nm); therefore, omitting the signal when DAPI is complexed with RNA (~ 500 nm). For the purpose of obtaining a well-defined cell cycle distribution profile, 100,000 cells (events) were acquired per sample and analyzed via Kaluza Flow Cytometry Software (Beckman Coulter). Each experimental point, as well as their corresponding controls, were processed concurrently and assessed in triplicate.
Gutierrez D.A., DeJesus R.E., Contreras L., Rodriguez-Palomares I.A., Villanueva P.J., Balderrama K.S., Monterroza L., Larragoity M., Varela-Ramirez A, & Aguilera R.J. (2019). A new pyridazinone exhibits potent cytotoxicity on human cancer cells via apoptosis and poly-ubiquitinated protein accumulation. Cell biology and toxicology, 35(6), 503-519.
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