Question

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Histology, immunohistochemistry, and electron microscopy.
To obtain fetal lung tissue, the fetuses were removed by
hysterotomy after lethal injection of pentobarbital sodium to
the dam. The chest of fetal animals was opened, and the
tissue was fixed with 4% paraformaldehyde at 4°C. Lungs
from postnatal animals were inflation fixed at 25 cmH2O
pressure via a tracheal cannula with the same fixative. TisTissue
was fixed overnight, washed in PBS, dehydrated through
a series of alcohols, and embedded in paraffin. Tissue sections
were stained for SP-B, proSP-B, thyroid transcription
factor (TTF)-1, proSP-C, CCSP, and 5-bromo-29-deoxyuridine
(BrdU) as previously described (10). For electron microscopy,
tissue was postfixed in 1% osmium tetroxide and evaluated
as previously described (18).
In situ hybridization. Expression of FGF-10 mRNA was
assessed by in situ hybridization using 35S-labeled riboprobes
as previously described for fetal and adult lungs (40),
the latter after inflation fixation at 25 cmH2O pressure.
Sense and antisense FGF-10 RNA probes were generated in
pGEM32. Tissue was hybridized overnight at 50°C. Slides
were coated with Kodak NTB-2 emulsion, exposed for 3–7
days, and developed with Kodak D19.

Answer 1

组织学，免疫组织化学，电镜。
要获得胚胎肺组织，胎儿被删除
子宫切开术后注射致死戊巴比妥钠以
大坝。胸部胎动物开幕式，并
组织固定， 4 ％甲醛在4 ℃的温度肺
从产后的动物通货膨胀率定为25 cmH2O
压力通过气管插管同固定。 TisTissue
固定一夜之间，在磷酸缓冲液洗涤，脱水通过
一系列的酒精，和石蜡包埋。切片
染色的螺一B proSP一B ，甲状腺转录
因子（转录因子） -1 ， proSP - C和CCSP ，以及5 -溴- 29 -脱氧
（尿嘧啶）正如先前所描述（ 10 ） 。对于电子显微镜，
组织postfixed在1 ％锇酸和评价
正如先前所描述（ 18 ） 。
原位杂交。表达的成纤维细胞生长因子- 10表达
评估原位杂交利用35S启动子标记riboprobes
正如先前所描述的胎儿和成人肺部（ 40 ） ，
后者扣除通货膨胀后的固定在25 cmH2O压力。
义成纤维细胞生长因子- 10的RNA探针产生
pGEM32 。杂交组织是一夜之间，在50摄氏度相片
被涂层与柯达的非关税壁垒- 2乳液，暴露3-7
天，和发达国家与柯达的D19 。