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Note that the Sec8 antibody recently described in [ 21] as not showing immunoreactivity at the fly NMJ, actually does so (and shows staining very similar to our antibody) when preparations are fixed in Bouin's fixative.
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Preparations were fixed with Quick Dip Fixative (Fronine Australia) and stained with Quick Dip (Fronine).
When antibodies against any of the glutamate receptor subunits were used, preparations were fixed 30 min in Bouin's fixative.
For these stainings, preparations were fixed for 15 min. in Bouin's fixative (Sigma, L'isle d'Abeau, France).
All cell preparations were fixed with Cytofix (BD Biosciences).
Single cell preparations were fixed in 4% paraformaldehyde in 0.1 M PBS for 20 minutes.
Cells were washed and all cell preparations were fixed with Cytofix (BD Biosciences).
Flattened cochlear preparations were fixed in 4% paraformaldehyde (in phosphate-buffered solution, PBS) for 30 minutes.
Whole-mount preparations were fixed by incubation with 0.4% PFA and washed in 0.1% Tween 20 in 1× PBS.
The preparations were fixed with 0.4% paraformaldehyde and mounted in a coverslip with an anti-bleaching solution (Dako, Denmark).
To count the number of macrophages, eosinophils and lymphocytes in BALF, cytocentrifuged preparations were fixed on slide and stained with Wright-Giemsa differential staining.
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