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, Vider les premiers jets, retour à la hotte : remplir 10mL de la seringue
, II Ajouter les cellules A partir de la solution intermédiaire :-préparer la cellule Mallassez : une goutte d'eau de chaque coté + lamelle tenue par les gouttes
Compter 10 cases, nombre de cellules = N *10 kC/mL, p.20 ,
,
, diluer la solution intermédiaire pour avoir 2kC/mL (*150 µL = 300 C/puits)
, mettre 150 µL/puits avec le multipipette parallèle ou une pipette normale
, Préparer l'échantillon spheroïde-préchauffer 3mL de milieu dans un Falcon
, mettre 1 mL de Polylisine dans une boîte de pétri et laisser 10 minutes ;-reprendre la Polylisine (ne pas la jeter)
, L'adhésion du sphéroïde sur le fond de la boîte de pétri se fait en 30 minutes environ