Sy3200

Manufactured by Sony

Sourced in United States

The SY3200 is a laboratory equipment product manufactured by Sony. It is a multi-purpose instrument designed to assist in various scientific and research applications. The core function of the SY3200 is to provide precise and reliable measurements and data analysis capabilities for users in the lab environment.

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Lab products found in correlation

Lsrfortessa, by BD (5 mentions) Pe conjugated anti hcd25 clone bc96, by BioLegend (3 mentions) Apc conjugatedanti hcd25 clone bc96, by BioLegend (3 mentions) Facscalibur, by BD (3 mentions) Facsaria, by BD (3 mentions) Pe conjugated anti hcd2, by BD (2 mentions) Facscanto, by BD (2 mentions) Moflo xdp, by Beckman Coulter (2 mentions) Lsrfortessa x 20, by BD (2 mentions) Apc conjugated streptavidin, by Thermo Fisher Scientific (1 mentions) Cytofix cytoperm solution, by BD (1 mentions) Liberase tl, by Merck Group (1 mentions) Propidium iodide, by BioLegend (1 mentions) Lsrii cytometer, by BD (1 mentions) Collagenase and hyaluronidase, by STEMCELL (1 mentions) Gentlemacs octo dissociator and tumor dissociation kit, by Miltenyi Biotec (1 mentions) Nf κb p65, by Cell Signaling Technology (1 mentions) Luciferase reporter kit, by Promega (1 mentions) Bradford method, by Bio-Rad (1 mentions) Celltrics 50 m filter, by Sysmex (1 mentions)

24 protocols using sy3200

Dual-population scRNA-seq of zebrafish melanoma

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In vitro samples were cultured under standard conditions with either ZMEL1-PRO (tdTomato) and -INV (EGFP) separately (individual culture) or mixed together in a 1:1 ratio for 11 days (co-culture). Cells were trypsinized, resuspended in DMEM supplemented with 2% FBS, and flow sorted using a SY3200 (Sony) for DAPI-negative pure ZMEL1-PRO and -INV populations prior to droplet-based scRNA-seq. For in vivo samples, adult casper zebrafish were transplanted with a 1:1 mixture of ZMEL1-PRO (tdTomato) and -INV (EGFP) cells as described above (Adult transplantation). Tumors were allowed to grow for 6 days (primary tumors) or 13 days (metastases). At experimental timepoint, tumors were surgically excised and minced with a fresh scalpel (primary tumors from n=6 fish; metastases from n=4 fish). Each sample was placed in a 15mL tube containing 3mL of 0.9X DPBS with 0.16 mg/mL of Liberase TL (Sigma #5401020001), incubated at room temperature for 15 minutes followed by trituration using a wide-bore P1000 (Fisher #2069G), and incubated for an additional 15 minutes. 500μL FBS was added and each sample was triturated again and passed through a 70μm cell strainer (Corning #352350). Samples were centrifuged 500g for 5 minutes, resuspended in DMEM supplemented with 2% FBS, and flow sorted using a SY3200 (Sony) for DAPI-negative pure ZMEL1-PRO and -INV populations prior to droplet-based scRNA-seq.

Campbell N.R., Rao A., Hunter M.V., Sznurkowska M.K., Briker L., Zhang M., Baron M., Heilmann S., Deforet M., Kenny C., Ferretti L.P., Huang T.H., Perlee S., Garg M., Nsengimana J., Saini M., Montal E., Tagore M., Newton-Bishop J., Middleton M.R., Corrie P., Adams D.J., Rabbie R., Aceto N., Levesque M.P., Cornell R.A., Yanai I., Xavier J.B, & White R.M. (2021). Cooperation between melanoma cell states promotes metastasis through heterotypic cluster formation. Developmental cell, 56(20), 2808-2825.e10.

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Flow Cytometry Analysis of Immune Cells

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Flow cytometric analyses were performed on a FACSCalibur or BD
LSRFortessa (BD Biosciences). Sorting was conducted on a Sony Sy3200 through
the Siteman Cancer Center Flow Cytometry Core Facility. Fluorescein
isothiocyanate (FITC)-conjugated anti-CD45R/B220 (clone RA3–6B2),
phycoerythrin (PE)-conjugated anti-CD43 (clone S7), FITC-conjugated
anti-CD43 (clone S7), PE-Cy7-conjugated anti-CD45/B220 (clone
RA3–6B2), allophycocyanin (APC)-conjugated anti-IgM (clone II/41),
APC-conjugated anti-hCD2, and PE-conjugated anti-hCD2 were purchased from BD
Biosciences. PE-conjugated anti-hCD25 (clone BC96) and APC-conjugated
anti-hCD25 (clone BC96) were purchased from BioLegend.

Soodgupta D., White L.S., Yang W., Johnston R., Andrews J.M., Kohyama M., Murphy K.M., Mosammaparast N., Payton J.E, & Bednarski J.J. (2019). RAG-Mediated DNA Breaks Attenuate PU.1 Activity in Early B Cells through Activation of a SPIC-BCLAF1 Complex. Cell reports, 29(4), 829-843.e5.

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Flow Cytometry Analyses and Sorting

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Flow cytometric analyses were performed on a FACSCalibur or BD LSRFortessa (BD). Sorting was conducted on a BD Aria (user-operated) or on a Sony Sy3200 through the Siteman Cancer Center Flow Cytometry Core Facility. FITC-conjugated anti-CD45R/B220 (clone RA3-6B2), phycoerythrin (PE)-conjugated anti-CD43 (clone S7), FITC-conjugated anti-CD43 (clone S7), PE-Cy7–conjugated anti-CD45/B220 (clone RA3-6B2), allophycocyanin (APC)-conjugated anti-IgM (clone II/41), PE-conjugated anti-CD40 (clone 1C10), and biotin-conjugated anti-CD40 (clone 3/23) were purchased from BD. APC-conjugated streptavidin was purchased from eBioscience. PE-conjugated anti-hCD25 (clone BC96) and APC-conjugated anti-hCD25 (clone BC96) were purchased from BioLegend. APC-conjugated anti-mCherry (clone 16D7) was purchased from Life Technologies. Staining for intracellular proteins was performed using Cytofix/Cytoperm solution (BD).

Bednarski J.J., Pandey R., Schulte E., White L.S., Chen B.R., Sandoval G.J., Kohyama M., Haldar M., Nickless A., Trott A., Cheng G., Murphy K.M., Bassing C.H., Payton J.E, & Sleckman B.P. (2016). RAG-mediated DNA double-strand breaks activate a cell type–specific checkpoint to inhibit pre–B cell receptor signals. The Journal of Experimental Medicine, 213(2), 209-223.

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Single-cell RNA-seq of Arabidopsis Root Cells

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Cells were isolated from cut roots using a short (1h-2h) cell wall digestion, followed by 3 filtrations through a 40µm screen. CFP positive protoplasts were sorted using FACS (either BD Aria, or Sony SY3200) into 96-well plates containing lysis buffer using gates to ensure a single cell per droplet (Figure S7A,B) Single cells were subject to cDNA synthesis, amplification, library preparation, read alignment and expression calling (Supplemental Experimental Procedures). Data was deposited in GEO (GSE74488). To derive cell identity scores, marker Spec scores were calculated as described previously (Efroni et al., 2015 (link)), and 579 tissue markers selected, checking first whether read depth affected cell identity or mixed identity calls (Figure S7C,D). To determine background ICI levels, we used ICI scores for QC, Columella and Epidermis\LRC in uncut, stele-derived cells as threshold values (Figure S7E, Supplemental Experimental Procedures). To produce the multidimensional scaling plot, we used expression of all 579 identity markers, which were z-normalized to reduce outlier effects and scaled using the cmdscale function in R.

Efroni I., Mello A., Nawy T., Ip P.L., Rahni R., DelRose N., Powers A., Satija R, & Birnbaum K.D. (2016). Root regeneration triggers an embryo-like sequence guided by hormonal interactions. Cell, 165(7), 1721-1733.

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Testis Cell Sorting via Flow Cytometry

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Testis cell suspensions were used for flow cytometry and fluorescence-activated cell sorting (FACS), essentially as described previously [11 (link)]. Briefly, cells were suspended (5–20 × 10⁶ cells/ml) in ice-cold Dulbecco PBS (DPBS) containing 10% FBS (DPBS + S), labeled with antibodies (Supplemental Table S1; Supplemental Data are available online at www.biolreprod.org), and subjected to flow cytometry using an LSRII cytometer (BD) or FACS using either a FACS Aria (BD) or SY3200 (Sony). Positive antibody labeling was determined by comparison to staining with isotype control antibodies (Supplemental Table S1). Positive ID4-EGFP epifluorescence was determined by comparison to testis cells from P6 Id4-eGfp⁻ littermates. For discrimination of dead cells, we used either propidium iodide (Biolegend) or LIVE/DEAD Fixable Violet or Near-IR Dead Cell Stain Kits (ThermoFisher Scientific).

Mutoji K., Singh A., Nguyen T., Gildersleeve H., Kaucher A.V., Oatley M.J., Oatley J.M., Velte E.K., Geyer C.B., Cheng K., McCarrey J.R, & Hermann B.P. (2016). TSPAN8 Expression Distinguishes Spermatogonial Stem Cells in the Prepubertal Mouse Testis. Biology of Reproduction, 95(6), 117.

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Breast Cancer Stem Cell Isolation

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Isolated primary tumors were mechanically dissociated by mincing with scalpels and suspended in Media 199 and single cell suspensions generated by incubation with collagenase and hyaluronidase (Stem Cell Technologies). Human tumor cells with DsRed label were then isolated using FACS on a SY3200 (Sony Biotechnology) flow cytometer. Secondary female, 5 week old, NOD/SCID mice were inoculated with 5,000, 1,000, or 200 cells for BT474 PTEN- LTT xenografts as described above. Tumor formation rate in secondary mice was assessed 8 weeks following implanting cells by direct palpitation and used to assess cancer stem cell frequency by extreme limiting dilution analysis.

Sun L., Burnett J., Gasparyan M., Xu F., Jiang H., Lin C.C., Myers I., Korkaya H., Liu Y., Connarn J., He H., Zhang N., Wicha M.S, & Sun D. (2016). Novel cancer stem cell targets during epithelial to mesenchymal transition in PTEN-deficient trastuzumab-resistant breast cancer. Oncotarget, 7(32), 51408-51422.

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Flow Cytometry Analysis of Immune Cells

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Adoptive T Cell Transfer in Tumor Models

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For T cell adoptive transfer experiments, CD3⁺ T cells from
RT (12 Gy) or sham-treated orthotopic tumors were FACS-sorted (SY3200, Sony) and
co-injected subcutaneously into mice in a 1:10 ratio with KPC-derived tumor
cells (1×10⁶) or Pan02 cells (5×10⁶; gift
of Daniel Meruelo, NYU). For PDA-macrophage co-culture assays, KPC-derived tumor
cells (5×10⁴) were irradiated at 12 Gy. One hour later,
FACS-sorted macrophages were added to the cancer cells (2:1 ratio). After 48
hours, macrophages were harvested and analyzed by flow cytometry.

Seifert L., Werba G., Tiwari S., Ly N.N., Nguy S., Alothman S., Alqunaibit D., Avanzi A., Daley D., Barilla R., Tippens D., Torres-Hernandez A., Hundeyin M., Mani V.R., Hajdu C., Pellicciotta I., Oh P., Du K, & Miller G. (2016). Radiation Therapy Induces Macrophages to Suppress Immune Responses Against Pancreatic Tumors in Mice. Gastroenterology, 150(7), 1659-1672.e5.

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Comprehensive B Cell Immunophenotyping

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Single-cell suspensions were obtained from spleen after erythrocyte lysis, and stained with fluorophore-conjugated anti-mouse or human antibodies (BD Pharmigen or Invitrogen) to detect B220 (RA3-6B2, 1/200), Fas (Jo2, 1/400), GL7 (1/200), hCD2 (S5.5, 1/200), immunoglobulin A (IgA) (1/200), immunoglobulin G1 (IgG1, A85-1, 1/400), CD19 (ID3, 1/400), CD25 (1/100) immunoglobulin M (IgM) (1/200), immunoglobulin D (IgD, 11-26, 1/200), CD21 (7G6, 1/200), CD23 (B3B4, 1/200) and CD93 (AA4.1, 1/200). Cell-cycle analysis was performed with Hoechst33342 staining of alive B cells for 60 min. Samples were acquired on LSRFortessa or FACSCanto instruments (BD Biosciences) and analysed with FlowJo software. For preparative flow cytometry FACSAria (BD Biosciences) or SY3200 (Sony) sorters were used.

Pérez-García A., Marina-Zárate E., Álvarez-Prado Á.F., Ligos J.M., Galjart N, & Ramiro A.R. (2017). CTCF orchestrates the germinal centre transcriptional program and prevents premature plasma cell differentiation. Nature Communications, 8, 16067.

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Xenograft Tumor Formation Assessment

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Isolated primary tumors were mechanically and enzymatically dissociated using gentleMACS Octo Dissociator and tumor dissociation kit (Miltenyi Biotec) according to manufacturer’s instructions. Human tumor cells with DsRed label were then isolated using fluorescent activated cell sorting on a SY3200 (Sony Biotechnology) flow cytometer. Secondary mice were inoculated with 10,000, 5,000, or 1,000 cells for BT474 xenografts and 5,000, 1,000, or 200 cells for BT474 PTEN⁻ LTT xenografts as described above. Tumor formation rate in secondary mice was assessed 9 weeks following implanting cells by direct palpitation.

Burnett J.P., Korkaya H., Ouzounova M.D., Jiang H., Conley S.J., Newman B.W., Sun L., Connarn J.N., Chen C.S., Zhang N., Wicha M.S, & Sun D. (2015). Trastuzumab resistance induces EMT to transform HER2+ PTEN− to a triple negative breast cancer that requires unique treatment options. Scientific Reports, 5, 15821.

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