5.6.1. Preparation and embedding of larval tissues
- Perform infection assays as described in section 4.1.
- Wash each larva for 5 seconds with 100 % ethanol.
- Fix larvae by overnight incubation with 4 % Roti Histofix (Roth, Karlsruhe) at 4°C at 20 rpm.
- Wash samples overnight with 1X PBS solution containing 6.8 % sucrose under the same conditions.
- Wash samples with 100 % acetone.
During the first five minutes, change acetone until the solution becomes clear.
- Incubate with the final change of acetone for 1 hour under the same conditions.
- Mix equal volumes of 100 % acetone and Technovit 8100 solution (Heraeus Kulzer, Wehrheim).
- Incubate samples for 2 hours with this solution under previous conditions.
- Perform infiltration and embedding using Technovit 8100 according to manufacturer´s instructions.
- Cut 4 µm-sections using a microtome, straighten them with warm sterile water on slides.
- Store slides at 4°C until further use.