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Diva software version 6

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DiVa Software, version 6.3, is a software application designed for use with BD laboratory equipment. The core function of this software is to facilitate the control and operation of the associated hardware devices.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (2 mentions) May gr nwald giemsa stain, by Merck Group (1 mentions) Apc conjugated cd235a, by BD (1 mentions) Facsaria, by BD (1 mentions) Sytox blue, by Thermo Fisher Scientific (1 mentions) Axioskop 40, by Zeiss (1 mentions) Facscanto 2, by BD (1 mentions) Gentlemacs tissue dissociator, by Miltenyi Biotec (1 mentions) Ariaiiiu, by BD (1 mentions) 7 aad, by BD (1 mentions) Facsaria 2, by BD (1 mentions) Rlt buffer, by Qiagen (1 mentions)

Close spelling variants of this product

Diva-version 6.0 software Diva software 6.0 Diva 6 software DIVA software

8 protocols using diva software version 6

1

Cell Cycle Analysis by Flow Cytometry

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Cell cycle phase distribution was analyzed at 24 h from post-starvation serum replacement by flow cytometry using PI staining (Sigma Aldrich). Briefly, Control or shFAK HCC cells were collected by trypsinization, washed with PBS, then fixed in a solution of a cold 4 : 1 methanol/acetone solution. Cells were first incubated with RNase A at +37 °C then stained with a solution containing 100 μg/ml PI, at +37 °C for 20 min. Stained nuclei were analyzed for DNA-PI fluorescence using a Becton Dickinson FACSCanto II flow cytometer (Becton-Dickinson, Milan, Italy). Resulting DNA distributions were analyzed for the proportions of cells in G0/G1, S phase, and G2/M phases of the cell cycle by DiVa Software, version 6.3 (Becton-Dickinson).
Gnani D., Romito I., Artuso S., Chierici M., De Stefanis C., Panera N., Crudele A., Ceccarelli S., Carcarino E., D'Oria V., Porru M., Giorda E., Ferrari K., Miele L., Villa E., Balsano C., Pasini D., Furlanello C., Locatelli F., Nobili V., Rota R., Leonetti C, & Alisi A. (2017). Focal adhesion kinase depletion reduces human hepatocellular carcinoma growth by repressing enhancer of zeste homolog 2. Cell Death and Differentiation, 24(5), 889-902.
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2

Apoptosis Assessment of SARS-CoV-2 Proteinase

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Apoptosis was assessed by Fluorescein-5-isothiocyanate (FITC) Annexin V Apoptosis Detection Kit I (code: 556547; Becton Dickinson-BD, Franklin Lakes, NJ, USA). Cells were seeded at a density of 100,000 cells/well in a 12-wells plate and were transfected with 800 nanograms of a plasmid containing SARS-CoV-2 (2019-nCoV) Papain-like proteinase/NSP3 Gene ORF cDNA clone expression plasmid or with pCMV3-untagged Negative Control Vector (all by Sino Biological Inc., Beijing, China). Briefly, cells were washed twice with cold PBS and resuspended in 1X Annexin Binding Buffer. Cells were then stained with 5 μL of FITC Annexin V and with 5 μM of Propidium Iodide (PI) for 15 min before analyzing. Acquisition and analysis were carried out on a FACSCanto II flow cytometer, using DiVa Software, version 6.3 (Becton, Dickinson and Company, Franklin Lakes, NJ, USA).
Crudele A., Smeriglio A., Ingegneri M., Panera N., Bianchi M., Braghini M.R., Pastore A., Tocco V., Carsetti R., Zaffina S., Alisi A, & Trombetta D. (2022). Hydroxytyrosol Recovers SARS-CoV-2-PLpro-Dependent Impairment of Interferon Related Genes in Polarized Human Airway, Intestinal and Liver Epithelial Cells. Antioxidants, 11(8), 1466.
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3

Cytospin and Flow Cytometry Analysis

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Cytospin preparations were obtained by cytocentrifugation (Shandon, Astmoor, UK) and stained with May-Grünwald-Giemsa stain (Sigma). Slides were observed with a Axioskop 40 microscope (Zeiss, Oberkochen, Germany) and images acquired with a CoolSNAP cf digital CCD digital camera (Photometrics Scientific, Tucson, AZ). For flow cytometry determinations of erythroid cells, samples were labeled with APC-conjugated CD235a and FITC-conjugated CD36 (Becton Dickinson Biosciences, Franklin Lakes, NJ). Dead cells were identified by staining with Sytox blue (1 μM) (Life Technologies, Waltham, MA). Cell fluorescence was analyzed with a FACSAria (Becton Dickinson Biosciences).
Cell Cycle Analyses: MNC (1.0–3.0×106) cultured in HEMA for 24, 48 and 72h with or without SB431542 (26 μM) were labelled with phycoerythrin-cyanin 7 (PE-Cy7)-conjugated CD34 or appropriate isotype controls (Becton Dickinson Biosciences), fixed overnight with paraformaldehyde (8% v/v) and then permeabilized with Triton X-100 (0.25% v/v). Permeabilized cells were labelled with phycoerythrin Ki-67 (Becton Dickinson Biosciences) and DAPI and analysed with the FACS Aria (Becton Dickinson Biosciences). Results were analysed with the Diva software version 6.1.3 (Becton Dickinson Biosciences).
Ceglia I., Dueck A.C., Masiello F., Martelli F., He W., Federici G., Petricoin EF I.I.I., Zeuner A., Iancu-Rubin C., Weinberg R., Hoffman R., Mascarenhas J, & Migliaccio A.R. (2016). Pre-clinical rationale for TGF-β inhibition as a therapeutic target for the treatment of myelofibrosis. Experimental hematology, 44(12), 1138-1155.e4.
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4

Apoptosis Quantification by Flow Cytometry

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Cells (1 × 106) were seeded in to six-well plates. After 24 h, culture medium was replaced with fresh complete medium and cells were allowed to grow for next 48 h. Subsequently, the extent of apoptosis was measured as previously described (Srivastava et al, 2012 (link); Arora et al, 2015 (link)). In brief, cells were harvested and stained with 7-amino-actinomycin (7-AAD) and PE Annexin V using commercially available kit (BD Pharmingen, San Diego, CA, USA) followed by flow cytometry on a BD-FACS Canto II (Becton-Dickinson, San Jose, CA, USA). Percentage of the apoptotic cells was calculated using DIVA software version 6.1.3 (Becton-Dickinson).
Srivastava S.K., Bhardwaj A., Arora S., Tyagi N., Singh S., Andrews J., McClellan S., Wang B, & Singh A.P. (2015). MicroRNA-345 induces apoptosis in pancreatic cancer cells through potentiation of caspase-dependent and -independent pathways. British Journal of Cancer, 113(4), 660-668.
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5

Single Cell Isolation and Staining

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Single cell suspensions were prepared from cultured cells using Puck’s EDTA, and from tumors using a gentleMACS Tissue Dissociator (Miltenyi Biotec, San Diego, CA, USA). Cells were stained with mAbs shown in Supplementary Table S1 in the dark for 45 minutes at 4° C, washed twice with PBS (without Ca++ and Mg++) containing 0.2% bovine serum albumin and 0.1% NaN3, and filtered through 50 μm mesh to remove clumps. 50,000 and 300,000 events were acquired per sample for cultured and tumor cells, respectively. Dead cells were excluded according to positive staining for DAPI. Debris was excluded by gating on forward and side scatter parameters. Doublets were excluded by gating on forward scatter area versus height. Data were acquired on an LSRII instrument with ultraviolet laser excitation of DAPI and analyzed using DIVA software version 6 (BD Biosciences, San Jose, CA, USA).
Webb M.W., Sun J., Sheard M.A., Liu W.Y., Wu H.W., Jackson J.R., Malvar J., Sposto R., Daniel D, & Seeger R.C. (2018). Colony Stimulating Factor 1 Receptor Blockade Improves the Efficacy of Chemotherapy against Human Neuroblastoma in the Absence of T Lymphocytes. International journal of cancer, 143(6), 1483-1493.
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6

Multiparameter Flow Cytometry Analysis

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The monoclonal antibodies αCD11c (clone B-ly6) and αCD14 (clone M5E2) were purchased from BD Biosciences Pharmingen. αCD8 (clone MEM-31) monoclonal antibody was obtained from Immuno Tools. αCD80 (clone 2D10), αCD86 (clone IT2.2), and αIFNγ (clone B27) antibodies were used from Biolegend. The αKi67 (clone SoIA15) antibody was purchased from ebioscience. Intracellular staining with αIFNγ and αKi67 was performed as described previously [17 (link)]. Data were acquired by using an LSR II instrument using DIVA software version 6 (BD Biosciences Pharmingen).
Scheffel F., Knuschke T., Otto L., Kollenda S., Sokolova V., Cosmovici C., Buer J., Timm J., Epple M, & Westendorf A.M. (2020). Effective Activation of Human Antigen-Presenting Cells and Cytotoxic CD8+ T Cells by a Calcium Phosphate-Based Nanoparticle Vaccine Delivery System. Vaccines, 8(1), 110.
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7

FACS Data Acquisition and Analysis

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Fluorescence-activated cell sorting (FACS) DIVA® software Version 6.1.3 (BD Biosciences; San Diego, CA) was used for data acquisition and analysis at the Covance Immunotoxicology Laboratory (Madison,WI) and Neumedicines Inc., Pasadena CA.
Gokhale M.S., Vainstein V., Tom J., Thomas S., Lawrence C.E., Gluzman-Poltorak Z., Siebers N, & Basile L.A. (2014). Single low-dose rHuIL-12 safely triggers multilineage hematopoietic and immune-mediated effects. Experimental Hematology & Oncology, 3, 11.
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8

Isolation and sorting of HFSCs

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Epidermal and dermal cell suspensions were incubated with the appropriate antibodies for 1 h at 4°C. Staining with 7-AAD (BD Bioscience) was used to exclude dead cells. Cell isolations were performed using a FACS AriaII or AriaIIIu equipped with Diva software version 6.1.3 (BD Bioscience). For RNA extraction of HFSCs, populations were sorted directly into Buffer RLT (Qiagen).
Morinaga H., Mohri Y., Grachtchouk M., Asakawa K., Matsumura H., Oshima M., Takayama N., Kato T., Nishimori Y., Sorimachi Y., Takubo K., Suganami T., Iwama A., Iwakura Y., Dlugosz A.A, & Nishimura E.K. (2021). Obesity accelerates hair thinning by stem cell-centric converging mechanism. Nature, 595(7866), 266-271.
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