Details for sorting your sample 1) Contact: name Group E-mail or phone 2) When is your preferred day? 3) The FACSAria II SORP has this Configuration: Laser UV Laser (355nm) Filter 450/50 530/30 Color DAPI Indo1 (blue) exam ple Hoechst 33258 Emerald 300 Violet Laser (405nm) Blue Laser (488nm) 450/50 525/50 525/50 Alexa 405 AmCyan FITC Pacific Blue Pacific Orange Alexa 488 GFP YellowGreen Laser (561nm) 695/40 585/15 610/20 660/20 PerCp PE PE-Texas Red PE-Cy5 mCherry mPlum PerCpCy5.5 DsRed 710/50 YGPerCpCy5-5 Red Laser (640nm) 780/60 670/14 APC PE-Cy7 PE-Cy5.5 Alexa 647 a) Put the used dyes in the right column of the table. b) You can only put in one color per column! E.g.: Rho 123: it is excited with the blue laser and the emission will be detected in the filter 525/50nm, so put Rho 123 in the column like FITC c) If the excitation and/or emission are unknown put the color name here: write the Excitation max. and Emission max. 730/45 form the protocol 780/60 APC-H7 Alexa 700 APCCy7 4) Sample, average cell size. cell size: 5) To optimize the sort time, answer the question, please. a) How many cells should be sorted? b) What is the percentage of your populations? (Note that you can’t sort more than 4 different populations at the same time!) d) What is your goal? 6) Does your sample need a certain temperature? % % % % Purification ? no 7) You should bring along: a) b) c) d) 8) Remark: Your stained/transfected and filtered sample. Tubes or pates for the cell to be sorted into, filled to >1/3 with growth medium. An unstained sample or for cells transfect with GFP or other fluorescent protein an untransfect and unstained sample. If you use more than one color, bring compensation controls with you (for that you can use your cells or beads).