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Animals were sacrificed by cervical dislocation, brains were immediately fixed in 2% buffered formaldehyde and paraffin embedded or immediately shock-frozen.
For histological analysis, brains were immediately fixed in formalin for 24 h and then embedded in paraffin according to standard procedures.
Fly heads of each genotype were opened and brains were immediately fixed with 4% paraformaldehyde (in 1× PBS) for 15 20 min, followed by dissection in 1× PBST (0.3% Triton X-100 in PBS) at room temperature.
Groups of nine mice were inoculated as described above, and on days 7, 14, and 21 post-infection, three mice from each group were euthanized by CO2 asphyxiation, and lungs and brains were immediately fixed in 10% neutral buffered formalin.
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Upon removal, the brain was immediately fixed in cold 4% paraformaldehyde in phosphate buffer (0.1 M, pH 7.4) and subsequently, small blocks of the hippocampal formation and adjacent cortex were obtained (10 × 10 × 10 mm) and post-fixed in the same fixative solution for 24 h at 4°C.
Mouse lung, brain, kidney, liver, spleen, placenta and fetus were immediately fixed in 10% formalin after sampling and embedded in paraffin.
In the cases where GFP/RFP fluorescence had to be kept, testes were immediately fixed as described for brain squashes (Sabino et al., 2011) except that the acetic acid step was omitted and the tissue gently squashed.
Samples were immediately fixed in 10%% neutral-buffered formalin.
These tissues were immediately fixed in 10% neutral buffered formalin.
Faecal samples were immediately fixed in SAF.
Thereafter, tissue cultures were immediately fixed.
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