Consider adding a higher concentration of FBS to the recovery media for sensitive samples. You can add roughly 10-20% of the receptacle volume (e.g. Multi-well plates (6, 12, 24, 48, 96, 384) (1-way sort only)Ĭoat the recovering tube/plate with media or PBS before arriving. 5 ml FACS tubes (ideally polypropylene as cells adhere less than in polystyrene) Add DNAse to your sample prior to sorting.Ĭells can be collected in various types of tubes or plates: This significantly increases cell clumping. Consider using cell dissociation products that maintain cell suspensions.įor sensitive cells: You can add 25 mM HEPES to stabilize the pH.įor samples with high cell death: DNA will be released in solution by the dead/dying cells. Note that cell tolerance for EDTA varies too much can kill your cells. Consider bringing extra buffer for dilution purposes.įor Sticky Cells: The EDTA concentration can be increased to 5 mM. Sterilize with a 0.2uM filter and store the solution at 4☌.īefore bringing your samples to be processed, filter them through a 40/70 uM mesh filter. Prepare the following buffer in which to suspend your samples after staining, prior to cell sorting:ġX Phosphate Buffured Saline (PBS) or Hanks Balanced Salt Solution (HBSS) (free of Ca2+ or Mg2+)ġ mM EDTA (can be removed for simple lymphocyte populations) The optimal threshold rate to achieve the best recovery/efficiency ratio will be the determining factor. Note that the nozzle size and sample concentration will affect the time of your sort. Please consult facility staff to decide the ideal conditions for your sort. 130 uM /16 PSI (Dissociated tumor cells/ large cell lines/ delicate- sensitive cells) 100 uM /20 PSI (Fibroblasts, cultured primary cells, medium to large cell lines, endothelial cells, adherent cell lines, transfected cell lines) 85 uM /45 PSI (Transduced-transfected splenocytes, thymocytes, bone marrow cells / B or T cell lines) 70 uM /70 PSI (Splenocytes/Thymocytes/PBMC/Whole blood/bone marrow) We offer 4 different sorting conditions for various cell types: The sort can be performed at different pressure and nozzle size. Monthly bill is capped at $650 for analyzers (LSC members only). *Life Science Complex (LSC) rate: McIntyre building labs, GCRC labs, Bellini building labs, Stewart building labs. The cytometers are accessible at any time for trained users.Ĭontact us for an instrument training, to schedule a sort or for technical advice on experimental design. for cell sorting services and user assistance. The facility is open Monday through Friday from 9:30 a.m. We also provide dongles for the analysis software FlowJo. Users or clients that won’t be doing regular acquisition and analysis can also ask for operator assisted data acquisition and analysis. We offer consultation for experiment design, data analysis and other technical assistance. We provide the mandatory training for the users to gain access to the cytometers. For a more complete description of the equipment (configurations and technical specificity), please consult our external webpage ( ). You can refer to the equipment section of this website to view the instrument available. An operator-based cell sorting service is also offered. Bring your data to be analyzed).The Flow Cytometry Core Facility is equipped with 5 different flow cytometers for various applications. This software provides open-ended modeling of flow cytometry histograms, and models for use with cell-tracking dye studies and synchronized cell lines (This software is only to be used by FCCS Staff. Allows FACS data analysis using a full set of region tools and gates (This software is only to be used by FCCS Staff. WinList 5.0: Cytometry software that allow the analysis of FCS files.Users can obtain a dongle required to use the software from the FCCS Facility. Users can download the latest FlowJo version for Windows or Mac. The FCCS Facility has two FlowJo licenses which are available to its users at no cost. FlowJo is a comprehensive analysis package specifically designed for handling list mode data generated by cytometers.
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