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Optische Sauerstoffmessung in der in vitro Zellkultur zur Bestimmung der orts- und zeitaufgelösten, diffusionslimitierten Sauerstoffkonzentrationen

Optische Sauerstoffmessung in der in vitro Zellkultur zur Bestimmung der orts- und zeitaufgelösten, diffusionslimitierten Sauerstoffkonzentrationen

          
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About the Book

Masterarbeit aus dem Jahr 2015 im Fachbereich Physik - Biophysik, Note: 1,0, Technische Universität München, Sprache: Deutsch, Abstract: Für die Aufrechterhaltung aller Lebensprozesse unseres Körpers wandelt jede Zelle kontinuierlich energiereiche Moleküle aus der Nahrung in chemisch speicherbare Energie um. Dazu braucht sie etwa 107 Sauerstoffmoleküle in der Sekunde. Die Kenntnis des Sauerstoffverbrauchs lässt Rückschlüsse auf den physiologischen Zustand der Zellen zu. Ebenso hängt die Physiologie der Zelle von dem in der Umgebung vorhandenen Sauerstoff ab. Deshalb wird der Sauerstoffgehalt in Zellkulturen bisher mit aufwändigen Methoden oder Berechnungen bestimmt. Für die heute angewandte Lebendzellmikroskopie wäre es von großem Vorteil den Sauerstoff in der Zelle oder in ihrer unmittelbaren Umgebung kontinuierlich zu messen. Es gibt bereits Messsysteme die mit phosphoreszierenden Farbstoffen eine sehr hohe räumliche und zeitliche Auflösung erreichen. Diese waren jedoch bislang nicht in Zellkultur einsetzbar. Deshalb wurde im Rahmen dieser Arbeit ein solches System für Lebendzellversuche adaptiert. Dazu wurde eine Dampfsterilisation der Beads geprüft. Es wurden verschiedene Methoden zur definierten Aufnahme der Beads in die Probe und zur Kalibrierung des Systems getestet und verglichen. In einem Kanal mit 30µl Volumen, in den metabolisierende Zellen eingesät wurden wird nach 24 Stunden eine zur Atmosphäre außerhalb des Objektträgers deutlich verringerte Sauerstoffkonzentration festgestellt. Die zeitabhängige Messung über 70 Stunden und etwa 12 Messpunkten auf 12mm Kanallänge zeigt einen schwachen lokalen Gradienten und eine starke Zeitabhängigkeit. Um die Korrelation der Zeitabhängigkeit mit der Zelldichte zu erforschen wurde die Sauerstoffkonzentration in Töpfchen mit 300µl Fassungsvolumen und drei verschiedenen Zelldichten 81 Stunden lang verfolgt. Der Verlauf der Sauerstoffkurve verändert sich charakteristisch mit der Zellkonzentration. Das ist ein klarer Hin


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Product Details
  • ISBN-13: 9783668005891
  • Publisher: Grin Verlag
  • Publisher Imprint: Grin Verlag
  • Height: 210 mm
  • No of Pages: 108
  • Series Title: German
  • Weight: 149 gr
  • ISBN-10: 3668005893
  • Publisher Date: 24 Jul 2015
  • Binding: Paperback
  • Language: German
  • Returnable: N
  • Spine Width: 7 mm
  • Width: 148 mm

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Optische Sauerstoffmessung in der in vitro Zellkultur zur Bestimmung der orts- und zeitaufgelösten, diffusionslimitierten Sauerstoffkonzentrationen
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Optische Sauerstoffmessung in der in vitro Zellkultur zur Bestimmung der orts- und zeitaufgelösten, diffusionslimitierten Sauerstoffkonzentrationen
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