{"id":159,"date":"2015-09-10T13:03:45","date_gmt":"2015-09-10T11:03:45","guid":{"rendered":"http:\/\/jcet.eu\/new\/?page_id=159"},"modified":"2023-02-23T17:01:25","modified_gmt":"2023-02-23T16:01:25","slug":"pracownia-farmakologii-doswiadczalnej-srodblonka-jcet","status":"publish","type":"page","link":"https:\/\/jcet.eu\/new\/o-jcet\/laboratoria-jcet\/pracownia-farmakologii-doswiadczalnej-srodblonka-jcet\/","title":{"rendered":"Farmakologia Do\u015bwiadczalna"},"content":{"rendered":"<h2><span style=\"color: #ff0000;\">Pracownia Farmakologii Do\u015bwiadczalnej \u015ar\u00f3db\u0142onka (JCET)<\/span><\/h2>\n<p><img decoding=\"async\" class=\"noimage alignleft\" src=\"http:\/\/jcet.eu\/images\/pfds_03.jpg\" alt=\"\" border=\"0\" \/><\/p>\n<p align=\"justify\">W Pracowni Farmakologii Do\u015bwiadczalnej \u015ar\u00f3db\u0142onka prowadzone s\u0105 badania <em>ex vivo<\/em> na izolowanych naczyniach (aorta, t\u0119tnica szyjna, t\u0119tnica krezkowa), a tak\u017ce na izolowanych narz\u0105dach (serce, w\u0105troba) z wykorzystaniem zwierz\u0119cych modeli dysfunkcji \u015br\u00f3db\u0142onka naczyniowego (np. niewydolno\u015bci serca, mia\u017cd\u017cycy, nadci\u015bnienia p\u0142ucnego, hypertriglicerydemii, cukrzycy czy chor\u00f3b zapalnych w\u0105troby). W wykorzystywanych modelach <em>in vitro<\/em>, <em>ex vivo<\/em> oraz <em>in vivo<\/em> badana jest, m.in. aktywno\u015b\u0107 naczynioprotekcyjnych, pro-zapalnych, pro-zakrzepowych przeka\u017anik\u00f3w \u015br\u00f3db\u0142onka (np. NO, PGI2, MCP-1, TM), kt\u00f3rych aktywno\u015b\u0107 odzwierciedla fenotyp czynno\u015bciowy i skuteczno\u015b\u0107 terapii. Pracownia dysponuje r\u00f3wnie\u017c bogatym repertuarem metod s\u0142u\u017c\u0105cych ocenie funkcji p\u0142ytek krwi, przebiegu proces\u00f3w zakrzepowych oraz szerokim zapleczem metod histologicznych, immunohistochemicznych oraz immunocytochemicznech, pozwalaj\u0105cych na wgl\u0105d w patologiczne zmiany na poziomie tkanek i narz\u0105d\u00f3w. Kierownikiem Pracowni PFD\u015a jest dr Marta Sm\u0119da.<\/p>\n<hr \/>\n<p>Najwa\u017cniejsz\u0105 aparatur\u0119 badawcz\u0105 Pracowni Farmakologii Do\u015bwiadczalnej \u015ar\u00f3db\u0142onka stanowi\u0105:<\/p>\n<p align=\"justify\"><strong>O\u015bmiokana\u0142owy zestaw k\u0105pieli kr\u0105\u017ck\u00f3w naczyniowych<\/strong> (Danish Myo Technology \u2013 DMT)<br \/>\nz zaawansowanym p\u00f3\u0142automatycznym systemem p\u0142ukania przeznaczony do bada\u0144 izolowanych naczy\u0144 krwiono\u015bnych o wi\u0119kszej \u015brednicy (2 mm \u2013 2 cm). Zestaw dw\u00f3ch czterokana\u0142owych miograf\u00f3w (DMT) do bada\u0144 izolowanych kr\u0105\u017ck\u00f3w naczyniowych naczy\u0144 krwiono\u015bnych o mniejszej \u015brednicy (np. aorta myszy o \u015brednicy 1 \u2013 2 mm lub t\u0119tniczka trzewna myszy o \u015brednicy ok. 100 \u03bcm).<\/p>\n<p align=\"justify\"><strong>Dwa miografy ci\u015bnieniowe<\/strong> (DMT) do bada\u0144 czynno\u015bci kaniulowanych ma\u0142ych naczy\u0144 krwiono\u015bnych (o \u015brednicy 60\u2013500 \u00b5m) w warunkach przep\u0142ywu przez \u015bwiat\u0142o naczynia zbli\u017conych do fizjologicznych.<\/p>\n<p align=\"justify\"><strong>Zestaw czterech miograf\u00f3w ci\u015bnieniowych<\/strong> (DMT) przeznaczony do d\u0142ugotrwa\u0142ych bada\u0144 czynno\u015bci kaniulowanych naczy\u0144 krwiono\u015bnych (o \u015brednicy od 60 \u00b5m do 2 mm).<\/p>\n<p align=\"justify\"><strong>Uk\u0142ad izolowanego pracuj\u0105cego serca<\/strong> (Hugo Sachs Elektronic-Harvard Apparatus, HSE-HA) umo\u017cliwiaj\u0105cy badanie czynno\u015bci pracuj\u0105cego serca myszy w warunkach ex vivo, w kt\u00f3rych serce jest perfundowane fizjologicznie poprzez lewy przedsionek i lew\u0105 komor\u0119.<\/p>\n<p align=\"justify\"><strong>System izolowanej perfundowanej w\u0105troby<\/strong> (Hugo Sachs Elektronik, Harvard Apparatus) to unikatowe narz\u0119dzie do biochemicznego i funkcjonalnego profilowania w\u0105troby oraz kr\u0105\u017cenia w\u0105trobowego w warunkach ex vivo. \u015aci\u015ble kontrolowane parametry perfuzji, mo\u017cliwo\u015b\u0107 wprowadzania dodatkowych zwi\u0105zk\u00f3w do uk\u0142adu (np. lek\u00f3w, inhibitor\u00f3w) czy pomiar st\u0119\u017cenia metabolit\u00f3w uwalnianych do prefuzatu pozwala na wykorzystanie tej metody r\u00f3wnie\u017c w badaniach farmakologicznych i farmakokinetycznych nowych lek\u00f3w.<\/p>\n<p align=\"justify\"><strong>Bie\u017cnia wysi\u0142kowa dla myszy z komor\u0105 Oxymax<\/strong> (Columbus Instruments). Urz\u0105dzenie jest przeznaczone do pomiar\u00f3w zu\u017cycia tlenu i wydalania CO2 przez organizm zwierz\u0119cia w toku \u015bci\u015ble kontrolowanego wysi\u0142ku fizycznego myszy na bie\u017cni i pozwala na pomiar wydolno\u015bci wysi\u0142kowej (VO2max).<\/p>\n<p align=\"justify\"><strong>XYZYK<\/strong> (XYZYK), sprz\u0119t zaprojektowany specjalnie na potrzeby JCET, s\u0142u\u017cy do biochemicznej oceny parametr\u00f3w p\u0142ytek krwi ex vivo.<\/p>\n<hr \/>\n<p><u>Pok\u00f3j biochemiczny<\/u><\/p>\n<p align=\"justify\"><strong>Analizator biochemiczny ABX Pentra 400<\/strong> (Horiba Medical) pozwala metodami spektrofotometrycznymi lub potencjometrycznymi na zautomatyzowany pomiar 55 podstawowych parametr\u00f3w surowicy lub osocza.<\/p>\n<p align=\"justify\"><strong>Analizator pHOx Ultra<\/strong> (Nova Biomedical) przeznaczony jest do pomiar\u00f3w parametr\u00f3w gazometrycznych krwi (PO2, PCO2) pe\u0142nej oksymetrii laboratoryjnej (m.in. pomiar zawarto\u015bci O2-Hb, CO-Hb, Met-Hb), oraz st\u0119\u017ce\u0144 podstawowych elektrolit\u00f3w krwi.<\/p>\n<p align=\"justify\"><strong>Stacja robocza Bioplex<\/strong> (BIO-RAD) jest urz\u0105dzeniem wykorzystuj\u0105cym nowoczesn\u0105 technologi\u0119 oznaczania substancji biologicznych z wykorzystaniem przeciwcia\u0142 zwi\u0105zanych z magnetycznymi kulkami polistyrenowymi, rozdzielanymi na frakcje metod\u0105 zbli\u017con\u0105 do cytometrii przep\u0142ywowej. Urz\u0105dzenie pozwala na oznaczenie nawet 100 r\u00f3\u017cnych bia\u0142ek, peptyd\u00f3w, fragment\u00f3w DNA RNA i innych moleku\u0142 w jednej pr\u00f3bce krwi o obj\u0119to\u015bci 50 \u00b5l.<\/p>\n<p align=\"justify\"><strong>Automat Gemini<\/strong> (Stratec Biomedical Systems) umo\u017cliwia w pe\u0142ni zautomatyzowane wykonanie wszystkich etap\u00f3w procedury ELISA. Urz\u0105dzenie potrafi kontrolowa\u0107 proces pipetowania trzema r\u00f3\u017cnymi metodami. Ma dwa standardowe czytniki do test\u00f3w ELISA (w tym jeden fluorescencyjny).<\/p>\n<p align=\"justify\"><strong>Mikrowaga kwarcowa z dyssypacj\u0105 energii<\/strong> (QCM-D, Quartz Crystal Microbalance with Dissipation monitoring) to nanoczu\u0142y cztero-kana\u0142owy system przep\u0142ywowy, wykorzystywany do ilo\u015bciowej oraz jako\u015bciowej oceny oddzia\u0142ywa\u0144 p\u0142ytek krwi z powierzchni\u0105 sensor\u00f3w, jak i efekt\u00f3w tych oddzia\u0142ywa\u0144 na biomechanik\u0119 p\u0142ytek krwi.<\/p>\n<hr \/>\n<p><u>Pok\u00f3j histologiczny i mikroskopowy<\/u><\/p>\n<p align=\"justify\"><strong>Kriostat elektromechaniczny z systemem utrzymywania czysto\u015bci w komorze<\/strong> (Leica CM 1950), rotacyjny mikrotom manualny (Sakura Accu-Cut SRM), stacja do zatapiania w parafinie (Thermo Scientific Microm EC 350-2), procesor tkankowy w systemie karuzelowym (Leica TP 1020), automatyczna stacja do barwienia preparat\u00f3w (Leica Autostainer XL).<\/p>\n<p align=\"justify\"><strong>Mikroskop fluorescencyjny Axio Imager.A2<\/strong> (Carl Zeiss) oraz odwr\u00f3cony mikroskop fluorescencyjny Axio Observer.D1 (Carl Zeiss) z zestawem filtr\u00f3w dla trzech barwnych kana\u0142\u00f3w, mo\u017cliwo\u015bci\u0105 obserwacji w kontra\u015bcie interferencyjnym DIC i PlasDIC; wyposa\u017cone w kamery monochromatyczne i kolorowe.<\/p>\n<p align=\"justify\"><strong>Stacja mikroskopii wirtualnej oparta o zautomatyzowany mikroskop Olympus BX51<\/strong> do skanowania i archiwizowania preparat\u00f3w histologicznych w postaci obraz\u00f3w o du\u017cej rozdzielczo\u015bci. Oprogramowanie skanera umo\u017cliwia udost\u0119pniane obraz\u00f3w w trybie online.<\/p>\n<p align=\"justify\"><strong>System skriningowy ScanR<\/strong>, sk\u0142adaj\u0105cy si\u0119 z odwr\u00f3conego, w pe\u0142ni zautomatyzowanego mikroskopu fluorescencyjnego (Olympus IX81), obudowanego inkubatorem z systemem pozwalaj\u0105cym na d\u0142ugotrwa\u0142e utrzymywanie mikro\u015brodowiska w\u0142a\u015bciwego dla hodowli kom\u00f3rkowej (Life Imaging Services). Ca\u0142o\u015b\u0107 tworzy unikatowe po\u0142\u0105czenie mikroskopu fluorescencyjnego z cytometrem obrazowym, pozwalaj\u0105ce na analiz\u0119 fenotypu kom\u00f3rek w warunkach fizjologicznych w czasie rzeczywistym.<\/p>\n<p align=\"justify\"><strong>Analizator metaboliczny Seahorse XFe96<\/strong> (Seahorse BioScience) umo\u017cliwia pomiary szybko\u015bci konsumpcji tlenu odzwierciedlaj\u0105cej oddychanie mitochondrialne oraz szybko\u015bci zewn\u0105trzkom\u00f3rkowego zakwaszenia odzwierciedlaj\u0105c\u0105 glikoliz\u0119 w \u017cywych kom\u00f3rkach, modelach kom\u00f3rkowych 3D oraz skrawkach tkanek. Szybko\u015b\u0107 konsumpcji tlenu mo\u017ce by\u0107 mierzona tak\u017ce w izolowanych mitochondriach lub permeabilizowanych kom\u00f3rkach. 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