The invention relates to an mRNA in-situ hybridization technology, in particular to a method suitable for bastard
halibut gonad frozen section mRNA in-situ hybridization. The method includes the steps that bastard
halibut gonad is immobilized with an excess PFA solution with the concentration of 4% and permeated with an excess
methanol solution with the concentration of 100% in sequence and then is preserved for a long term; the sample which is immobilized and then preserved is subjected to rehydration and 20-30% saccharose
sedimentation, then is subjected to OCT embedding and frozen section preparing and is dried for 2-3 h at 55 DEG C, and a frozen section is obtained to serve as an in-situ hybridization sample; the sample is washed for 3-5 min with excess DEPC water to remove an OCT
embedding agent in the sample frozen section; the sample frozen section is subjected to washing,
proteinase K digestion and immobilization, then is subjected to pre-hybridization and then is mixed with a hybridization solution containing a bastard
halibut RNA probe at 50-70 DEG C for overnight hybridization; after hybridization treatment, washing,
antibody incubation, rewashing, light-proof color development, termination color development, re-immobilization and section encapsulation are carried out, and thus expression localization of bastard halibut
gonad related genes at the
mRNA level is achieved. By means of the method, the problem that direct gonad in-situ hybridization of bastard halibut gonadal tissue is difficult is solved, and expression localization of bastard halibut related genes in gonad at the
mRNA level can be clearly described.