Moflo XDP高速细胞分选技巧与参数优化研究

doi:10.5246/jcps.2020.05.033

中国药学(英文版) ›› 2020, Vol. 29 ›› Issue (5): 355-363.DOI: 10.5246/jcps.2020.05.033

Moflo XDP高速细胞分选技巧与参数优化研究

孙玉芳¹, 郑晨², 徐波^1*

1. State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2. Beckman Coulter Commercial Enterprise (China) Co., LTD, Beijing 100004, China

收稿日期:2020-02-18 修回日期:2020-04-05 出版日期:2020-05-31 发布日期:2020-04-12
通讯作者: Tel.: +86-10-82805217, E-mail: xubo@bjmu.edu.cn

Research on Moflo XDP high speed cell sorting techniques and parameter optimization

Yufang Sun¹, Chen Zheng², Bo Xu^1*

1. 北京大学医学部药学院天然药物及仿生药物国家重点实验室, 北京 100191
2. 贝克曼库尔特商贸(中国)有限公司, 北京 100004

Received:2020-02-18 Revised:2020-04-05 Online:2020-05-31 Published:2020-04-12
Contact: Tel.: +86-10-82805217, E-mail: xubo@bjmu.edu.cn

摘要/Abstract

摘要：

本文主要探讨调校Moflo XDP高速分选设备进行细胞分选实验的方法, 优化仪器进入最佳运行状态, 实现高活性高效率分选。以荧光微球快速校准主液流与激光束的位置, 根据不同大小喷嘴调整振幅和频率, 找到液滴震荡最佳断点, 调整四路侧液流稳定性, 计算液滴延迟, 实现设备稳定的维持状态, 进而快速实现样本的高活性高效率分选。鞘液桶与液路中没有气泡是实现稳定分选的关键, 断点和液滴延迟的准确调校, 是提高分选效率和细胞质量的关键。

关键词: 细胞分选, 气泡, 维持, CV条件优化

Abstract:

This article aims to discuss strategies to operate Moflo XDP high-speed sorting equipment for cell sorting and optimize the instrument to run in the best operating state. Using fluorescent microspheres to quickly calibrate the position of the main liquidflow and the laser beam, adjust the amplitude and frequency according to different size CytoNozzles to find the best breakoff point, and adjust the four-way side flow to achieve the stable maintaining of the equipment, so as to quickly sort sample at high-speed. The absence of air bubbles in the sheath liquid barrel and the liquid path is the key to achieving a stable sorting-maintaining in the sorting process. The accurate calculation of the breakoff point and drop delay is the key to improve sorting efficiency and cell activity.

Key words: Cell sort, Bubble, Maintaining, CV optimization

中图分类号:

R917

Supporting:

孙玉芳, 郑晨, 徐波. Moflo XDP高速细胞分选技巧与参数优化研究[J]. 中国药学(英文版), 2020, 29(5): 355-363.

Yufang Sun, Chen Zheng, Bo Xu. Research on Moflo XDP high speed cell sorting techniques and parameter optimization[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(5): 355-363.

参考文献

[1] Laboratory medicine branch of Chinese Medical Association; Clinical test center of the Ministry of Health; Editorial board of Chinese Journal of laboratory medicine. Suggestions for clinical application of flow cytometry. Chin. J. Lab. Med. 2013, 36, 1064-1073.

[2] Saeys, Y.; van Gassen, S.; Lambrecht, B.N. Computational flow cytometry: helping to make sense of high-dimensional immunology data. Nat. Rev. Immunol. 2016, 16, 449-462.

[3] Oliver, O.; Philipp, R.; Alexander, M.; Stefan, G.; Christoph, H.; Daniel, K.; Salvatore, G.; Stefano, P.; Andrew, E.; Angela, J.; Manja, W.; Nicole, T.; Ulrich, F.; Mansfeld, J.; Elisabeth, F.; Jochen, G. Real-time deformability cytometry: on-the-fly cell mechanical phenotyping. Nat. Methods. 2015, 12, 199-202.

[4] Ekaterina, I.; Mark, G.; Taras, I.; Alexander, V.; Mazen, A.; Mustafa, S.; Dmitry, A.; Vladimir, P. In vivo acoustic and photoacoustic focusing of circulating cells. Sci. Rep. 2016, 6, 21531.

[5] Shi, Y.P.; Chong, Y.B.; Wang, Q. Selection and evaluation of high-end flow cytometer. Chin. Med. Equip. J. 2010, 31, 122-123, 125.

[6] Zhang, Y.X.; Zhang, L.Y.; Zhao, Q. Current status and prospects of medical application of flow cytometry. Chin. Modern Med. 2019, 26, 28-30.

[7] Chen, Z.B; Cao, X.T. Flow cytometry: principle, operation and Application. China Sci. Press. 2010, 1-3.

[8] Osborne, G.W. Recent advances in flow cytometric cell sorting. Methods Cell Biol. 2011, 102, 533-556.

[9] Du, L.Y. MoFlo high - performance cell sorter produced by DakoCytomation. Mod. Instruments. 2004, 10, 46-47.

[10] Liu, Y.; Wang, L.T.; Liu, F.; Ran, H.Y. Discussion on the experience of flow cell separation and optimization. Tech. Mana. 2018, 1, 55-56.

[11] Arnold, L.W.; Lannigan, J. Practical issues in high-speed cell sorting. Curr. Protoc. Cytom. 2010, Chapter 1, Unit 1.24.1-Unit 1.24.30.

[12] Zhou, M.; Zhang, Q.S.; Bao, M.Z. Study on parameter adjustment of flow cytometrer in cell sorting experiment. Med. Equip. 2016, 29, 58-61.

Moflo XDP高速细胞分选技巧与参数优化研究

Research on Moflo XDP high speed cell sorting techniques and parameter optimization

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