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However, the embryo wash fluid from two mares was still positive for EAV after all 10 washes and the two trypsin treatments, and one embryo was positive for EAV.
Next they were washed in embryo wash buffer (0.7% NaCl, 0.05% Triton X-100) and fixed by vigorous shaking in a 1∶1 heptane/methanol mix for 30s.
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Briefly, the hippocampi were obtained from E17 mouse embryos, washed in Ca2+/Mg2+-free HBSS, digested with 0.25% trypsin for 15 minutes at 37°C and dissociated.
At day 7 (blastocyst) and day 8 (fully expanded blastocyst) of embryo culture, a total of 5 8 embryos (washed in PBS) were freezed in liquid nitrogen.
(B ) Application of NA leads to aggregation of melanosomes in kif5aa mutant embryos; washing out of the drug results in their re-expansion.
Subsequently, embryos were washed three times in washing buffer and incubated in 1 200 goat anti-rabbit Alexa Fluor 488 (Molecular Probes) solution in blocking buffer overnight at 4 °C.
Secondary antibody was aspirated and embryos were washed in 4 15 minute PBT washes at RT.
Antibody in blocking buffer was aspirated and embryos were washed in 4 15 minute PBT washes at RT.
Embryos were washed twice in PBST for 15 min, at room temperature, followed by five 1 hr washes.
Embryos were washed through three drops of M2 medium.
Embryos were washed through two more dishes of M2 medium.
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