双相水凝胶组织固定联合脂肪酶浸泡对小鼠肝组织透明化的影响

Effects of biphasic hydrogel tissue fixation combined with lipase immersion on mice hepatic tissue transparency

目的:探讨双相水凝胶组织固定联合脂肪酶浸泡对小鼠肝组织透明化的影响。方法:取10只昆明小鼠的肝组织,利用双相水凝胶进行固定,切取1mm厚的肝组织10片,将其随机分为十二烷基硫酸钠(SDS)组和SDS+脂肪酶组。SDS组采用SDS清除液进行被动脂质清除9d,SDS+脂肪酶组先采用SDS清除液被动脂质清除5d,随后浸泡于脂肪酶溶液中4d。两组于清除前、清除后第1天至第9天均进行透明灰度值测定,于脂质清除完毕后予以细胞核DAPI染色检测标记深度。结果:在被动脂质清除过程中,第1天至第5天肝组织薄片的透明灰度值逐渐提升,但之后肝组织片清除速度缓慢,在第10天的清除仍然是不透明的;在第7、第9天,两组相对透明灰度值比较,差异有统计学意义(P<0.01);SDS+脂肪酶组DAPI染色深度达115μm,而SDS组DAPI染色深度达60μm,SDS+脂肪酶组DAPI渗透深度是SDS组的1.92倍。结论:双相水凝胶组织固定联合脂肪酶浸泡脂质清除技术可以快速实现小鼠肝组织透明化,为肝组织生物学研究提供技术支持。

Objective： To investigate the biphasic hydrogel tissue fixation combined with lipase immersion on mice hepatic tissue transparency.Methods： The liver tissues of mice were fixed with biphasic hydrogels and then cut into 1 mm slices.The slices were divided into sodium dodecyl sulfate （SDS） group （cleared with SDS for 9 days） and SDS＋lipase group （cleared with SDS for 5 days then immersed in lipase solution for 4 days）.The transparent gray value was measured before clearance and from day 1 to day 9 after clearance.After the lipid removal was finished,the nuclear DAPI staining was performed to detect the labeling depth.Results： In the process of passive lipid clearance,the transparent gray scale values of liver tissue slices increased gradually from day 1 to day 5,but after that,the clearance rate of liver tissue slices was slow,and the clearance was still opaque on the 10th day.The relative transparent gray values between the two groups were significantly different （ P 〈0.01）.The depth of DAPI staining in SDS＋lipase group and SDS group was 115 μm and 60 μm,respectively.The depth of DAPI penetration in SDS＋lipase group was 1.92 times higher than that in SDS group.Conclusion： Biphasic hydrogel tissue fixation combined with lipase passive clearance technology could rapidly obtain the transparency of hepatic tissue in mice.It may provide a promising technical support in the liver tissue biology research field.