FACS Request Fluorescence-Activated Cell Sort Request If you are a member of the CIT and would like to have cells isolated for you using a cell sorter (FACS), please fill out the form below to request a machine and a time to be reserved for you. Name Lab* Email Sorter to Reserve*BD FACSAria IIIBio-Rad S3eNozzleWhich nozzle should be inserted into the machine? The rule is that the size of your cells of interest should not exceed one-fifth of the nozzle diameter. Cells that deviate from an ideal spheroid shape are more susceptible to shear induced damage and cell death. If you have a small cell, switching to the larger (85um nozzle) often results in better viability. Nozzle size guideline: 5-10um = 70um nozzle, 11-16um = 85um nozzle & >16um = 100um nozzle.70um Nozzle85um Nozzle100um NozzleActive PMTsPlease select the PMT(s) that will need to be active during your sort. 405nm -> 450/40 (Pacific Blue) 405nm -> 510/50 (Pacific Orange) 488nm -> 530/30 (FITC) 488nm -> 585/42 (PE) 488nm -> 616/23 (PE-Texas Red) 488nm -> 695/40 (PerCP-Cy5.5) 488nm -> 780/60 (PE-Cy7) 633nm -> 660/20 (APC) 633nm -> 780/60 (APC-Cy7) Active PMTsPlease select the PMT that will need to be active during your sort. Fluorochrome choices are listed from high to low intensity. FL1 (525/30; VioBright FITC, BB515, Alexa Fluor 488, FITC, DyLight 488, YFP, Fluorescein, GFP, Cy2) FL2 (586/25; PE) FL3 (615/25; PE-Vio615, PE-CF594, PE-Dazzle594, PE-eFluor610, ECD, DyLight594, PE-Texas Red) FL4 (655LP; PE-Vio770, PerCP-eFluor710, PerCP-Vio700, BB700, PE-Cy5.5, PerCP-Cy5.5, PE-Cy7) Sample Priority*Please know that we receive rare human samples and may need to reschedule your sort with <24 hours notice. (1) nPOD (2) Rare Human Clinical Samples (3) Human Clinical Samples (4) Other Samples Patient/Sample #Please let use know if there is a patient or sample ID numbers associated with the samples. Date*The sorters are heavily utilized machines. Reservations requests should be made for two weeks or more from today's date. Month Day Year Start Time*A general guideline is that a sorter can handle 3x10^7 cells per hour. Hours : Minutes AM PM AM/PM End Time* Hours : Minutes AM PM AM/PM When possible, sorts should be begun early enough so that they are completed by 4p.m.
