对地高辛（ｄｉｇｏｘｉｇｅｎｉｎ，ＤＩＧ）标记的反义ＲＮＡ探针用于原位杂交的技术问题进行研究。结果表明：①杂交标本取材后应在１ｈ内放入固定液，时间过长，杂交信号明显降低；②探针工作浓度以０．４ｍｇ／Ｌ为宜，过高或过低影响杂交背景或阳性率；③防止ＲＮａｓｅ的污染是技术成败的关键之一；④杂交温度和时间以４３℃，１６ｈ为宜；⑤防止脱片是原位杂交的基本技术需要。 The technical problems of digoxigenin (DIG) labeled antisense RNA probe for in situ hybridization were studied. The results showed that: (1) After fixation, the immobilized liquid should be placed within 1 h after the sample was taken, and the hybridization signal was obviously decreased. (2) The working concentration of the probe should be 0.4 mg / L, too high or too low to affect the hybridization background or positive Rate; ③ prevent RNase pollution is one of the key to the success of the technology; ④ hybridization temperature and time to 43 ℃, 16h is appropriate; ⑤ prevent delamination is the basic technical needs of in situ hybridization.