{"id":2309,"date":"2014-04-29T21:15:00","date_gmt":"2014-04-30T01:15:00","guid":{"rendered":"https:\/\/www.med.unc.edu\/flowcytometry\/instrumentation-2\/general-lab-rules\/"},"modified":"2022-06-08T14:29:41","modified_gmt":"2022-06-08T18:29:41","slug":"general-lab-rules","status":"publish","type":"page","link":"https:\/\/www.med.unc.edu\/flowcytometry\/instrumentation\/general-lab-rules\/","title":{"rendered":"General Lab Rules and Collection Tubes"},"content":{"rendered":"<p style=\"text-align: justify\"><strong><span style=\"color: #000000\">Food and Drinks<\/span><\/strong><span style=\"color: #000000;line-height: 1.43em\"><br \/>\nFood and Drinks are not allowed in any of the Flow Cytometry Core Facility laboratories.<\/span><\/p>\n<p style=\"text-align: justify\"><span style=\"color: #000000\"><b>Logging in Instruments&#8217; Computers<\/b><\/span><br \/>\n<span style=\"color: #000000\">Enter your ONYEN and Password. Be sure you have selected to login to the AD domain.<\/span><\/p>\n<p style=\"text-align: justify\"><span style=\"color: #000000\"><b>Logging off <b>Instruments&#8217; Computers<\/b><\/b><\/span><br \/>\n<span style=\"color: #000000\">Please log off the machine when you are finished. You may be charged for extra time if you do not.<\/span><\/p>\n<p style=\"text-align: justify\"><strong><span style=\"color: #000000\">Sample Tubes Requirements<\/span><\/strong><\/p>\n<ul style=\"text-align: justify\">\n<li><span style=\"line-height: 1.43em;color: #000000\"><span style=\"line-height: 1.43em;color: #000000\">LSR II \/ LSRFortessa &#8211; <a href=\"https:\/\/us.vwr.com\/store\/catalog\/product.jsp?catalog_number=60819-273\">12x75mm (5mL) polystyrene plastic tubes<\/a><\/span><\/span><\/li>\n<li><span style=\"line-height: 1.43em;color: #000000\">ImageStreamX MkII &#8211; 1.5mL Eppendorf tubes<\/span><\/li>\n<li><span style=\"line-height: 1.43em;color: #000000\">MoFlo &#8211; 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes<\/span><\/li>\n<li><span style=\"line-height: 1.43em;color: #000000\">MoFlo XDP &#8211; 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes<\/span><\/li>\n<li><span style=\"line-height: 1.43em;color: #000000\">FACSAria II &#8211; 12x75mm (5mL) polystyrene or polypropylene plastic tubes to 50ml tubes and 1mL microtube size<\/span><\/li>\n<\/ul>\n<p style=\"text-align: justify\"><b style=\"color: #000000;line-height: 1.43em\">Sort Collection Tubes<\/b><span style=\"color: #000000;line-height: 1.43em\"><br \/>\nUse either 12x75mm (5mL) polypropylene plastic tubes or 12x120mm (15mL) polypropylene.<\/span><strong style=\"color: #000000;line-height: 1.43em\"><span style=\"text-decoration: underline\"><em><br \/>\nNote<\/em><\/span><\/strong><span style=\"color: #000000;line-height: 1.43em\">: Polystyrene tubes are not ideal as they accumulate charges during sorting which can result in losses of sorted cells.<\/span><\/p>\n<ul style=\"text-align: justify\">\n<li><span style=\"line-height: 1.43em;color: #000000\">For the 12x75mm tubes place ~1ml of media in the tube.<\/span><\/li>\n<li><span style=\"line-height: 1.43em;color: #000000\">For the 12x120mm tubes place ~2ml of media in the tube.<\/span><\/li>\n<li><span style=\"color: #000000\"><span style=\"line-height: 1.43em\">When using the 70\u00b5m tip we can collect ~3&#215;10<\/span><sup style=\"line-height: 1.43em\">6<\/sup><span style=\"line-height: 1.43em\"> cells per tube.<\/span><\/span><\/li>\n<li><span style=\"color: #000000\"><span style=\"line-height: 1.43em\">When using the 100\u00b5m tip we can place about ~1&#215;10<\/span><sup style=\"line-height: 1.43em\">6<\/sup><span style=\"line-height: 1.43em\"> cells per tube.<\/span><\/span><\/li>\n<li><span style=\"color: #000000\"><span style=\"line-height: 1.43em\">When using the <\/span><span style=\"line-height: 1.43em\">12x120mm<\/span><span style=\"line-height: 1.43em\"> tubes increase the numbers above by 3 fold.<\/span><\/span><\/li>\n<\/ul>\n<p style=\"text-align: justify\"><strong style=\"line-height: 1.43em\"><span style=\"color: #000000\">Instrument Clogs<\/span><\/strong><span style=\"color: #000000\"><br \/>\nFilter your samples prior to running on any instrument (<a href=\"\/flowcytometry\/resources-2\/cell-filtration-options\">Filter Options<\/a>).<\/span><\/p>\n<p style=\"text-align: justify\"><span style=\"color: #000000\">DNA from lysed cells in the medium can cause cells to clump. DNAse I (Sigma Cat. No. D-4513) in the presence of magnesium chloride (Sigma Cat. No. M-2670) will help reduce cellular aggregation. Treat cells for 15 to 30 minutes in a solution of 100 \u00b5g\/mL DNAse and 5 mM MgCl2 in HBSS at room temperature. Wash the cells once in the presence of 5 mM MgCl2 in HBSS. Gently suspend the cells in Stain Buffer (BSA) containing MgCl2 and 25-50 \u00b5g\/mL DNAse (as a maintenance dose). DNAse I requires a concentration of at least 1 mM magnesium to work effectively, although 5 mM is optimal. Note: It is important to minimize the presence of dead cells during this procedure, since actin released from dead cells irreversibly inhibits DNAse I. <\/span><\/p>\n<p style=\"text-align: justify\"><strong style=\"line-height: 1.43em\"><span style=\"color: #000000\">Instrument Cleaning<\/span><\/strong><span style=\"color: #000000;line-height: 1.43em\"><br \/>\nFollow the cleaning listed in the Quick Reference or affixed to the instrument. Please clean any spills and throw any used pipet tips, tube, tube caps, aluminum foil&#8230;in the trash.<\/span><\/p>\n<p style=\"text-align: justify\"><strong><span style=\"color: #000000\">Time Management<\/span><\/strong><span style=\"color: #000000;line-height: 1.43em\"><br \/>\nMake sure you stop your run before the end of your scheduled time to perform cleaning and in order not to delay the next user.<\/span><\/p>\n<p style=\"text-align: justify\"><strong><span style=\"color: #000000\">Data Location<\/span><\/strong><a style=\"line-height: 1.43em\" href=\"\/flowcytometry\/core-access-scheduling-2\/server-access\"><br \/>\nServer Access<\/a><\/p>\n<p style=\"text-align: justify\"><span style=\"color: #000000\">\u00a0<\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Food and Drinks Food and Drinks are not allowed in any of the Flow Cytometry Core Facility laboratories. Logging in Instruments&#8217; Computers Enter your ONYEN and Password. Be sure you have selected to login to the AD domain. Logging off Instruments&#8217; Computers Please log off the machine when you are finished. You may be charged &hellip; <a href=\"https:\/\/www.med.unc.edu\/flowcytometry\/instrumentation\/general-lab-rules\/\" aria-label=\"Read more about General Lab Rules and Collection Tubes\">Read more<\/a><\/p>\n","protected":false},"author":79063,"featured_media":0,"parent":2226,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"layout":"","cellInformation":"","apiCallInformation":"","footnotes":"","_links_to":"","_links_to_target":""},"class_list":["post-2309","page","type-page","status-publish","hentry","odd"],"acf":[],"_links_to":[],"_links_to_target":[],"_links":{"self":[{"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/pages\/2309","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/users\/79063"}],"replies":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/comments?post=2309"}],"version-history":[{"count":0,"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/pages\/2309\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/pages\/2226"}],"wp:attachment":[{"href":"https:\/\/www.med.unc.edu\/flowcytometry\/wp-json\/wp\/v2\/media?parent=2309"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}