Mammary tumors were dissected and minced, and then incubated in digestion medium (DMEM/F12 with 2% Penicillin/Streptomycin, 0.1 mg ml−1 Gentamicin, 0.6% Nystatin, 2 mg ml−1 Collagenase A, 0.096 mg ml−1 Hyaluronidase) at 37°C with shaking for 1-1.5 hour. After digestion, the cells/tissues were treated sequentially with 0.25% trypsin/EDTA (37°C, 2 minutes), 5 mg ml−1 dispase with DNaseI (0.1mg ml−1, Sigma, St. Louis, MO) (37°C, 5 minutes), cold red blood cell (RBC) lysis buffer (on ice, 2-3 minutes). Between each treatment step, cells/tissues were washed with 1x PBS with 5% FCS. After treatment with the RBC lysis buffer, cells/tissues were filtered through 40 μm cell strainer and washed with 1x PBS/5% FCS, to obtain single cell suspension [49 (link)]. Flow cytometric (FACS) analysis was performed with an Accuri C6 analyzer (BD Biosciences, San Jose, CA) and analyzed with CFlow (BD Biosciences), or with a DXP11 flow cytometer and analyzed with FlowJo. FACS cell sorting was performed with a FACSAria sorter (BD Biosciences).
Dxp11 flow cytometer
Manufactured by BD
The DXP11 flow cytometer is a laboratory instrument designed for cell analysis. It utilizes the principles of flow cytometry to rapidly detect and measure various characteristics of cells or other particles suspended in a fluid stream.
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Lab products found in correlation
2 protocols using dxp11 flow cytometer
1
Dissociation and FACS Isolation of Mammary Tumor Cells
2
Dissociation and FACS Isolation of Mammary Tumor Cells
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Mammary tumors were dissected and minced, and then incubated in digestion medium (DMEM/F12 with 2% Penicillin/Streptomycin, 0.1 mg ml−1 Gentamicin, 0.6% Nystatin, 2 mg ml−1 Collagenase A, 0.096 mg ml−1 Hyaluronidase) at 37°C with shaking for 1-1.5 hour. After digestion, the cells/tissues were treated sequentially with 0.25% trypsin/EDTA (37°C, 2 minutes), 5 mg ml−1 dispase with DNaseI (0.1mg ml−1, Sigma, St. Louis, MO) (37°C, 5 minutes), cold red blood cell (RBC) lysis buffer (on ice, 2-3 minutes). Between each treatment step, cells/tissues were washed with 1x PBS with 5% FCS. After treatment with the RBC lysis buffer, cells/tissues were filtered through 40 μm cell strainer and washed with 1x PBS/5% FCS, to obtain single cell suspension [49 (link)]. Flow cytometric (FACS) analysis was performed with an Accuri C6 analyzer (BD Biosciences, San Jose, CA) and analyzed with CFlow (BD Biosciences), or with a DXP11 flow cytometer and analyzed with FlowJo. FACS cell sorting was performed with a FACSAria sorter (BD Biosciences).
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