Frozen sections of chicken, frog (Xenopus laevis) and zebrafish hearts were used in immunofluorescence microscopy as previously described [7].
Immunofluorescence microscopy was carried out on frozen sections of chicken, frog (Xenopus laevis) and zebrafish (Danio rerio) hearts to determine whether Xin co-localization with β-catenin at the intercalated discs is evolutionarily conserved.
Sections of chicken embryo were fixed in 4 % paraformaldehyde at 4 °C overnight and were permeabilized with 0.5%% (v/v) Triton X-100 in PBS.
This was also confirmed in sections of E4 chicken embryos, where strong CYR61 signal was present in ECs of larger blood vessels (data not shown).
To enrich for motoneurons, only the ventral sections of the chicken spinal cord were excised into a Ca2+/Mg2+-free solution, mildly trypsinized (E8, 0.05%, 30 min; E11, 0.2% for 40 min), dissociated by trituration, and plated onto poly-d-lysine-coated glass coverslips.
We also carried out immunofluorescence experiments on frozen sections of the chicken spleens.
The pH values of specific sections of the chicken GI tract are the following: crop 4.5, proventriculus 4.4, gizzard 2.6, duodenum 5.7 to 6.0, jejunum 5.8, ileum 6.3, colon 6.3, caeca 5.7, and bile 5.9 [ 21].
Immunostaining was performed on free-floating brain sections using combinations of chicken anti-GFP (Invitrogen), rabbit anti-Iba1 (WAKO, Germany) and rabbit anti-neutrophil serum (SJC, kindly provided by Drs Daniel Anthony and Sandra Campbell, University of Oxford, Oxford, UK).
(The menu includes a section of "breaded chicken" dishes).
The chromatin fibers in the cryo-tomograms of Hela nuclei in situ appear similar to those observed in the chemically fixed and high-pressure frozen Hela nuclei, and in accordance with the conventional mechanical cryo-sections of isolated chicken erythrocyte nuclei (Scheffer et al. 2011).
A conventional transversal histological section of the chicken cornea is also shown for comparison (Fig. 1(a)).
