{"id":64,"date":"2018-01-08T10:14:45","date_gmt":"2018-01-08T10:14:45","guid":{"rendered":"http:\/\/confolab.sav.sk\/ovsb\/?page_id=64"},"modified":"2023-09-25T13:02:14","modified_gmt":"2023-09-25T13:02:14","slug":"konfokalna-mikroskopia","status":"publish","type":"page","link":"https:\/\/confolab.sav.sk\/ovsb\/laboratoria\/konfokalna-mikroskopia\/","title":{"rendered":"Konfok\u00e1lna mikroskopia"},"content":{"rendered":"<h2>Laserov\u00fd skenovac\u00ed konfok\u00e1lny fluorescen\u010dn\u00fd syst\u00e9m TCS SP2 AOBS (Leica Microsystems) s invertovan\u00fdm mikroskopom DMIRE-2 (Leica)&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;<button onclick=\"goBack()\">Nasp\u00e4\u0165<\/button> <\/h2>\n<p><script>\t \t \nfunction goBack() { window.history.go(-1); }\t \t \n<\/script><\/p>\n<h3>Kontakt: RNDr. Alexandra Zahradn\u00edkov\u00e1 ml., PhD.<\/h3>\n<h3 style=\"line-height:1.3\">Vybudovan\u00e9 s podporou projektov <a href=\"https:\/\/confolab.sav.sk\/ovsb\/projekty\/ukoncene-projekty\/hhmi-55000343\/\">HHMI-55000343<\/a>, <a href=\"https:\/\/confolab.sav.sk\/ovsb\/projekty\/ukoncene-projekty\/nih-firca-ro3-tw005543-2\/\">NIH FIRCA RO3 TW005543\/2<\/a>, <a href=\"https:\/\/confolab.sav.sk\/ovsb\/projekty\/ukoncene-projekty\/lshm-ct-2005-018833\/\">LSHM-CT-2005-018833<\/a> a <a href=\"https:\/\/umfg.sav.sk\/infrastrukturne-projekty\/itms-26240120008\/\" >ITMS 26240120008<\/a><\/h3>\n<table border=\"0\" width=\"100%\" cellspacing=\"1\" cellpadding=\"2\">\n<tbody style=\"line-height:1.3\">\n<tr>\n<td colspan=\"2\">\n<p>Konfok\u00e1lny mikroskop Leica TCS SP2 je vyu\u017e\u00edvan\u00fd prim\u00e1rne na meranie lok\u00e1lnych v\u00e1pnikov\u00fdch sign\u00e1lov po\u010das aktiv\u00e1cie uvo\u013e\u0148ovania v\u00e1pnika v xt m\u00f3de, kedy jedna l\u00ednia je skenovan\u00e1 opakovane r\u00fdchlos\u0165ou a\u017e 2000 l\u00edni\u00ed za sekundu  (obr\u00e1zok v\u013eavo). Z\u00edskan\u00fd obraz poskytuje \u00fadaje o priestore (x) a \u010dase (t).  Pomocou synchroniz\u00e1tora je konfok\u00e1lny mikroskop spojen\u00fd so zostavou patch-clamp aparat\u00fary. Toto zapojenie n\u00e1m umo\u017e\u0148uje bunky kontrolovane stimulova\u0165 a ich pr\u00fadov\u00fa odpove\u010f a uvo\u013enenie v\u00e1pnika zaznamen\u00e1va\u0165 paralelne. Fluorescen\u010dn\u00e9 indik\u00e1tory v\u00e1pnika kombinovan\u00e9 s nefluorescen\u010dn\u00fdmi chel\u00e1tormi v\u00e1pnika zobrazuj\u00fa priestorovo oddelen\u00e9 lok\u00e1lne udalosti aj v pr\u00edpade, \u017ee k uvo\u013e\u0148ovaniu v\u00e1pnika doch\u00e1dza v celom  objeme bunky naraz.<\/p>\n<\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\"><img decoding=\"async\" loading=\"lazy\" class=\"\" title=\"riadkov\u00e9 zobrazenie v\u00e1pnikov\u00fdch hrotov v srdcov\u00fdch svalov\u00fdch bunk\u00e1ch potkana\" src=\"https:\/\/confolab.sav.sk\/sk\/images\/spikes.gif\" alt=\"riadkov\u00e9 zobrazenie v\u00e1pnikov\u00fdch hrotov v srdcov\u00fdch svalov\u00fdch bunk\u00e1ch potkana\" width=\"198\" height=\"99\" hspace=\"1\" vspace=\"1\" \/> <img decoding=\"async\" loading=\"lazy\" class=\"\" title=\"ozna\u010denie povrchovej membr\u00e1ny a t-tubulov s di-8-ANEPPSom\" src=\"https:\/\/confolab.sav.sk\/sk\/images\/ANEPPS.gif\" alt=\"ozna\u010denie povrchovej membr\u00e1ny a t-tubulov s di-8-ANEPPSom\" width=\"343\" height=\"99\" hspace=\"1\" vspace=\"1\" \/><\/td>\n<\/tr>\n<tr>\n<td colspan=\"2\">Konfok\u00e1lny mikroskop Leica TCS SP2 je samozrejme mo\u017en\u00e9 pou\u017e\u00edva\u0165 aj na meranie statick\u00fdch obrazov alebo pomal\u0161\u00edch javov v \u0161tandardnom xy m\u00f3de. Fluorescen\u010dn\u00e9 zna\u010dky sa pou\u017e\u00edvaj\u00fa na zobrazenie \u0161pecifick\u00fdch \u010dast\u00ed buniek, ako s\u00fa povrchov\u00e1 membr\u00e1na (obr\u00e1zok vpravo) a in\u00e9 organely. Obrazy je mo\u017en\u00e9 sn\u00edma\u0165 v s\u00e9ri\u00e1ch optick\u00fdch rezov umo\u017e\u0148uj\u00facich 3D rekon\u0161trukciu, ako aj v \u010dasov\u00fdch s\u00e9ri\u00e1ch. Pri spr\u00e1vnej kombin\u00e1cii fluorescen\u010dn\u00fdch indik\u00e1torov  v\u00e1pnika a bunkov\u00fdch \u0161trukt\u00far je mo\u017en\u00e9 korelova\u0165 z\u00e1znamy z oboch typov zobrazen\u00ed.<\/p>\n<h3>Zostava konfok\u00e1lneho mikroskopu Leica TCS SP2:<\/h3>\n<ul>\n<li>Invertovan\u00fd mikroskop DM IRE2<\/li>\n<li>Objekt\u00edvy 10x a 20x such\u00fd, 40x olejov\u00fd, 63x olejov\u00fd, 63x vodn\u00fd<\/li>\n<li>AOBS, AOTF<\/li>\n<li>Skener 1000 Hz<\/li>\n<li>2x PMT, 1x TLD detektory<\/li>\n<li>1x Ar laser (458, 476, <strong>488<\/strong>, 496, 514 nm)<\/li>\n<li>3x He\/Ne laser (543, 594, <strong>633 <\/strong>nm)<\/li>\n<\/ul>\n<\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n","protected":false},"excerpt":{"rendered":"<p>Laserov\u00fd skenovac\u00ed konfok\u00e1lny fluorescen\u010dn\u00fd syst\u00e9m TCS SP2 AOBS (Leica Microsystems) s invertovan\u00fdm mikroskopom DMIRE-2 (Leica)&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Nasp\u00e4\u0165 Kontakt: RNDr. Alexandra Zahradn\u00edkov\u00e1 ml., PhD. Vybudovan\u00e9 s podporou projektov&#8230;<span class=\"excerpt_more\"><a href=\"https:\/\/confolab.sav.sk\/ovsb\/laboratoria\/konfokalna-mikroskopia\/\">\u010d\u00edta\u0165 \u010falej &raquo;<\/a><\/span><\/p>\n","protected":false},"author":1,"featured_media":0,"parent":18,"menu_order":20,"comment_status":"closed","ping_status":"closed","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/pages\/64"}],"collection":[{"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/comments?post=64"}],"version-history":[{"count":13,"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/pages\/64\/revisions"}],"predecessor-version":[{"id":2476,"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/pages\/64\/revisions\/2476"}],"up":[{"embeddable":true,"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/pages\/18"}],"wp:attachment":[{"href":"https:\/\/confolab.sav.sk\/ovsb\/wp-json\/wp\/v2\/media?parent=64"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}