Cells were dissociated into single cells with Accutase (STEMCELL Technologies, Cambridge, UK) for 10 minutes and subsequently fixed with 1% paraformaldehyde for 20 minutes at room temperature and stained with primary and secondary antibodies in PBS with 0.2% Triton X‐100 and 0.5% BSA. Data were collected on a FACSCaliber or FASCanto flow cytometer (Becton Dickinson, Franklin Lakes, New Jersey) and analyzed using FlowJo. Fluorescence‐activated cell sorting (FACS) gating was based on the corresponding isotype or secondary only antibody control.
Fascanto
Manufactured by BD
Sourced in United States
The FASCanto is a flow cytometer designed for multi-parameter analysis of cells and particles. It is capable of measuring various cellular characteristics, including size, granularity, and the expression of specific proteins or molecules on the cell surface or within the cell. The FASCanto provides researchers with a tool for advanced cell analysis and sorting applications.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using fascanto
1
Cell Dissociation and Flow Cytometry
2
Single-Cell Fixation and Staining Protocol
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Cells were dissociated into single cells with Accutase (Thermo Fisher Scientific) for 10 minutes, and subsequently fixed with 1% paraformaldehyde for 20 minutes at room temperature and stained with primary and secondary antibodies in phosphate buffered solution (PBS) with 0.2% Triton X‐100% and 0.5% bovine serum albumin (BSA). Data were collected on a FACSCaliber or FASCanto flow cytometer (Becton, Dickinson and Company, Franklin Lakes, NJ) and analyzed using FlowJo. Fluorescence‐activated cell sorter (FACS) gating was based on the corresponding isotype or secondary only antibody control.
3
Immunophenotyping Blood and Bone Marrow
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4
Apoptosis Assay for BPA and NP in LNCaP Cells
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LNCaP cells treated with 100 µM BPA or 50 µM NP for 6 h were washed twice with PBS. After that, they were incubated with 5 µL of FITC-Annexin-V and 5 µL of propidium iodide for 15 min at room temperature in the dark. Then, 400 µL of binding buffer was added and analyzed by flow cytometry (FAScanto, BD Biosciences, Franklin Lakes, NJ, USA) and 10,000 gated events were acquired in each sample. All data were analyzed with software FCS express V4.0 (De Novo Software, Los Angeles).
5
Quantifying Cellular Apoptosis by Flow Cytometry
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Following treatment, cell apoptosis was analyzed by flow cytometry (FASCanto; BD Biosciences) by staining with FITC-labeled Annexin V and propidium iodide (PI). MinWID 2.9 (BD Biosciences) was used for data acquisition and analysis. The summation of both early (Annexin V+ and PI-) and late (Annexin-V+ and PI+) apoptotic cells was used to determine the percentage of cells undergoing apoptotic death.
6
Cell Cycle Analysis by Flow Cytometry
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After treatment, LNCaP and A2780 cells were fixed in 70% ethanol. The cells were then pelleted and washed once with PBS. Then, the pellet was dissolved in a buffer containing 0.1% sodium citrate, 0.3% Triton X-100 (Sigma), 50 mg/mL propidium iodide, and 50 mg/mL RNase A (Invitrogen, Carlsbad, CA, USA). The samples were then analyzed by a flow cytometer (FAScanto, BD Biosciences, Franklin Lakes, NJ, USA). Ten thousand gated events were acquired in each sample. All data were analyzed with software FCS express V4.0 (De Novo Software, Los Angeles, CA, USA).
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