Vorbereitung von gefrorenem FISH-Gewebe

Standardprotokoll

  • 5 &mgr;m dicke Schnitte auf silanisierten oder positiv geladenen Objektträgern zuschneiden.
  • Bei Raumtemperatur 15 bis 30 Minuten trocknen lassen.
  • In 4 %igem Formaldehyd in PBS bei 0 °C 15 Minuten fixieren.
  • Mit PBS spülen, mit dem 2-N-HCl-Schritt aus dem Protokoll des Paraffin-Vorbehandlungskits beginnen und das Protokoll bis zum Ende befolgen.

Alternatives Protokoll

  • Die Schnitte wie zuvor zuschneiden und trocknen lassen.
  • In carnoyschem Fixiermittel (Methanol:Essigsäure im Verhältnis 3:1) 5 Minuten fixieren.
  • Unmittelbar vor dem Protease-Schritt (Spülpuffer, 5 Minuten) mit dem Protokoll des Paraffin-Vorbehandlungskits beginnen.

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