Optimization and application of microwave assisted rapid ultrathin section staining
10.3760/cma.j.cn112866-20200724-00215
- VernacularTitle:微波辅助超薄切片快速染色技术的优化与应用
- Author:
Hongli FENG
1
;
Hao XIAO
;
Haibin YU
;
Wenzhe HOU
;
Jingdong SONG
;
Hong TAO
Author Information
1. 中国疾病预防控制中心病毒病预防控制所 传染病预防控制国家重点实验室，北京 100052
- Keywords:
Ultrathin section;
Microwave;
Rapid staining;
Transmission electron microscope
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(5):556-561
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To optimize key parameters based on microwave assisted rapid staining technique for ultrathin section and establish a rapid ultrathin section preparing method .Methods:Ultrathin sections were stained respectively with 1% uranium acetate (UA) and lead citrate (LC) for different duration at various microwave power using microwave tissue processor. Then transmission electron microscope (TEM) photographs of cellular ultrastructure were taken and analyzed. The optimized single staining parameters were decided and combined to investigate the optimal microwave assisted UA and LC double staining conditions. The rapid staining effects of ultrathin sections were verified in different viruses including human adenovirus 5 (HAd5), herpes simplex virus (HSV), H1N1 influenza virus, enterovirus 71(EV-A71)human infected cells samples.Results:The optimized microwave assisted rapid ultrathin section staining parameters are: UA for 30 s and LC for 20 s at power 200 W or UA for 30 s and LC for 30 s at 300 W using microwave tissue processor.1∶6 dilution of original LC concentration could still work well through microwave assistance. The parameters can be extended and applied to domestic microwave ovens, and the optimized staining parameters are UA for 30 s and LC for 30 s at 320 W.Conclusions:The optimized parameters of microwave assisted rapid ultrathin section staining were obtained and can be applied in not only cell samples but also different virus ultrathin sections.
