[1]赵方新,张 烜,陆景坤,等.斑马鱼肝脏组织油红O染色方法比较优化[J].医学信息,2022,35(17):27-30,36.[doi:10.3969/j.issn.1006-1959.2022.17.006]
　ZHAO Fang-xin,ZHANG Xuan,LU Jing-kun,et al.Comparison and Optimization of Oil Red O Staining Methods in Liver Tissue of Zebrafish[J].Journal of Medical Information,2022,35(17):27-30,36.[doi:10.3969/j.issn.1006-1959.2022.17.006]
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斑马鱼肝脏组织油红O染色方法比较优化()
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医学信息[ISSN:1006-1959/CN:61-1278/R]

卷:
35卷
期数:
2022年17期
页码:
27-30,36
栏目:
论著
出版日期:
2022-09-01

文章信息/Info

Title:
Comparison and Optimization of Oil Red O Staining Methods in Liver Tissue of Zebrafish
文章编号:
1006-1959（2022）17-0027-05
作者:
赵方新; 张 烜; 陆景坤; 等
（内蒙古医科大学基础医学院医学实验中心，内蒙古 呼和浩特 010110）
Author(s):
ZHAO Fang-xin; ZHANG Xuan; LU Jing-kun; et al.
(Medical Experiment Center of Basic Medical College,Inner Mongolia Medical University,Hohhot 010110,Inner Mongolia,China)
关键词:
斑马鱼; 油红O染色; 肝脏; 冷冻切片
Keywords:
Zebrafish; Oil red O staining; The liver; Frozen sections
分类号:
R446.8
DOI:
10.3969/j.issn.1006-1959.2022.17.006
文献标志码:
A
摘要:
目的 通过设置不同染色步骤、条件，探索适合处理斑马鱼肝脏的冷冻切片油红O染色方法。方法 取3月龄斑马鱼肝脏OCT样品，设置6、8、10、12 μm四种不同厚度切片各两片，其他各组切片厚度均为8 μm，采用不同的处理方法处理两张切片。染色后置于显微镜下观察视野内组织完整度、背景情况、橘红色脂滴的染色面积、位置以及深染的细胞和位置形态。结果 以4%多聚甲醛固定、10%~30%蔗糖梯度脱水，切片厚度设置８~10 μm，室温晾干1 h，甲醛（4%）固定10 min，油红O染色10 min，60%异丙醇分化2 s，ddH2O浸洗浸泡5 s后，苏木素染色3 min，ddH2O浸泡30 s去浮色，1%盐酸酒精分化2 s，流水冲洗2 min返蓝，甘油明胶封片为条件进行油红O染色的效果较好，斑马鱼肝组织组织完整、背景清晰，脂滴染色效果较好。结论 优化的斑马鱼油红O染色方法可以较为清晰地显示肝脏脂滴，可辨别细胞核轮廓位置，进而对斑马鱼肝病模型中的脂肪变性情况进行定位与半定量的分析。
Abstract:
Objective To explore the suitable oil red O staining method for frozen section of zebrafish liver by setting different staining steps and conditions.Methods The OCT samples of 3-month-old zebrafish liver were taken. Four different thickness slices ( 6, 8, 10, 12 μm ) were set, and the thickness of other slices was 8 μm. After staining, the tissue integrity, background, staining area and location of orange lipid droplets, and deeply stained cells and location morphology were observed under a microscope.Results Zebrafish liver tissue was fixed with 4% paraformaldehyde, dehydrated with 10%-30% sucrose gradient, slice thickness was set at 8-10 μm, dried at room temperature for 1 h, fixed with formaldehyde (4%) for 10 min, stained with oil red O for 10 min, differentiated with 60% isopropanol for 2 s, soaked in ddH2O for 5 s, stained with hematoxylin for 3 min, soaked in ddH2O for 30 s to remove the floating color, differentiated with 1% hydrochloric acid alcohol for 2 s, washed with water for 2 min to return to blue, and sealed with glycerin gelatin. The effect of oil red O staining was better, the liver tissue of zebrafish was complete, the background was clear, and the effect of lipid droplet staining was better.Conclusion The optimized zebrafish oil red O staining method can clearly show the liver lipid droplets and identify the position of the nuclear contour, and then locate and semi-quantitatively analyze the steatosis in the zebrafish liver disease model.

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更新日期/Last Update: 1900-01-01