THE INFLUENCING ELEMENTS AND OPTIMIZATION   OF X-gal STAINING METHOD IN THE STUDY  OF TRANSPLANTATION

VernacularTitle:移植研究中X-gal染色的影响因素及优化条件
Author: Qi ZHANG ; Jinhong MENG ; Na LUO ; Jingwen LI ; Gong JU
Publication Type:Journal Article
Keywords: galactosidase; LacZ gene; Transplantation; Brain; Rat; Mouse
From: Acta Anatomica Sinica 1955;0(03):-
CountryChina
Language:Chinese
Abstract: Objective To study the influencing elements in x-Gal staining method and optimize the reactive conditions so that nonspecific background can be eliminated and grafted exogenous cells carrying LacZ gene can be discerned correctly. Methods C17 2 cells (carrying lzcZ gene) were injected into the right lateral ventricle both in the adult and newborn animals. After one week they were perfused using two methods, then the slices were stained at different pH and incubating time respectively. The X-gal positive cells in hippocampus were counted under light microscope. Results Background staining in this method has close correlation with the species and age of the host animals and it decreases when pH is higher or incubating time is shorter. The results are the best when pH 9 5 and the incubating time is 1*!h.Conclusion The reliability of X-gal staining method depends on optimization of several parameters, including pH, incubating time, perfusion etc. It is necessary to establish the correspondent controls.;