![]() ![]() Filtering often causes loss of cells, but is unavoidable for a trouble-free sort. Rinsing with clear buffer will dilute your cell concentration, but help to avoid loss due to clogging. We have nylon mesh filter tubes in the lab that we will use at cost ($1/tube) as needed if we notice any clumping in the tube you bring. Pour your sample through the mesh prior to sorting. This also reduces the risk of clogging.Ĭell samples must be filtered through 40 um nylon mesh to remove clumps of tissue, etc. For larger cell types like cell lines we run the machine at 25psi on the 100um tip. Please note that for smaller cell types like red blood cells, we recommend 60psi with the 70um tip. The actual speed of sorting your selected cells will depend on the percentage of positive cells and the concentration of the sample. Your sorted population depends on the percentage of "good" cells in the selected populations. ![]() Please keep in mind that event rate is measuring every particle detected by the machine, including any debris not gated out. (It's always easier to dilute rather than concentrate a sample.) A summary of our recommendations for sorting prep are here: Cell Sorting FAQ Sort ratesġ2,000 and 25,000 events per second are the maximum sorting event rates for the 100um and 70um tips respectively. Bring extra buffer so the we can dilute the sample if it's too concentrated. The sample can be suspended in the buffer that works best with your cell type to keep them alive during the sort. We recommend a concentration around 5 million cells/mL. Stream turbulence will cause lower purities in your sorted tubes. Clumps cause instability in the flow tip or can clog, causing a delay in continuing your sort and extending the length of your appointment. Cell PrepĬell samples must be in a single-cell suspension. You only need to fill this out once unless the cell type or preparation procedure changes. E-mail it or bring it with you for the discussion or before your sort. For example: What kind of cells are you sorting? How many cells do you need? What concentration should the sample be? How much time is needed? Which fluorochromes?įill out the pre-sort form as best as you can. For brand new experiments, often users will meet with staff to discuss the purpose and goal of the experiment and offer any advice for preparation or what settings on the machine may work best. Do not pour anything down the drain unless specifically directed to do so by a staff member.Į-mail Desa Rae Abrams to schedule a new sorting appointment or for questions about sorting.Assume all chemicals are hazardous, regardless of label, handle with caution.All users are required to complete laser safety training from EH&S and provide a copy of your certificate of completion to the Flow Cytometry staff as well as e-mailing a copy to All fixatives, solvents and media must be handled in a hood with caution. ![]() #Flowjo departmental license freeIt is recommended that all persons working in a laboratory attend the chemical safety training seminar, a free course given by Environmental Health and Safety at Penn State. There is a zero tolerance policy for unsafe or careless handling of chemicals or equipment in the facility. The only exception is the Astrios cell sorter which can run up to BSL-2+ materials since it is housed in a bio-safety cabinet. If they are potentially infectious, or human cell lines they should be fixed before running them on any of our machines. All samples run on our machines should be BSL-1 materials. However we still have chemicals like concentrated ethanol, bleach, and potentially toxic dyes like Propidium Iodide and DAPI, so gloves should be worn when working around any potential chemical or biological hazard in the lab. There are not many chemical hazards in the Flow Cytometry Core. You will need to bring a copy or e-mail staff a copy of the training certificate you receive at the end as well as e-mail a copy to safety trainingĪll persons working in a laboratory should attend the chemical safety training seminar, a free course given by Environmental Health and Safety at Penn State. Required safety trainingĪll users must complete Penn State EH&S Laser Safety training prior to being trained on any flow cytometer. We currently do not train users on our cell sorters. #Flowjo departmental license plusBilling for training will include the equipment time, plus and additional staff training rate ($34.64/hr). Training is usually divided into several 1-2 hour sessions each encompassing specimen preparation and basic equipment use. We provide training on an individual basis. Various University safety courses are also recommended or required. We offer training on equipment use and sample preparation. ![]()
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