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Author(s):: YU Zixuan ; ZHANG Xiaomin ; ZHOU Xinli; School of Medical Instrument and Food Engineering，University of Shanghai for Science and Technology，Shanghai 200093，China

Keywords:: Microdroplet spray; Spray parameters; Nitrogen flow rate; Injection speed; Suspension concentration; Receiving position; Needle type; Vitrification

摘要:: 微滴喷射玻璃化是指将细胞悬液离散成微滴后直接喷入液氮中进行玻璃化保存。该方法可有效提升降温速率，提高细胞玻璃化保存效果。微滴喷射过程中，喷射参数对微滴喷射后的细胞活性有较大影响。本研究对HepG2细胞进行微滴喷射实验，研究氮气流速、悬浮液注射速度、悬浮液浓度、微滴接收位置和针头型号等五个喷射参数对微滴大小及喷射后细胞存活率的影响。实验结果表明，当氮气流速为3.2 L/min、悬浮液注射速度为200 μL/min、悬浮液浓度为10% Me2SO、接收位置为距离喷嘴6 cm处且针头型号为30G时，微滴喷射后HepG2细胞活性最高。优化后的喷射参数可以作为其他小体积细胞微滴喷射玻璃化保存的参考。

Abstract:: Micro-droplet spray vitrification is the process of separating cell suspension into micro-droplets and injecting directly into liquid nitrogen for vitrification, this method can effectively increase the cooling rate. In the process of micro-droplet spray, the spray parameters have great influence on the cell activity after micro-droplet spraying. In this paper, HepG2 cells were subjected to micro-droplet spray, the effects of five spray parameters，such as nitrogen flow rate, suspension injection speed, suspension concentration, receiving position and needle type on the droplet size and cell viability were investigated. The results showed that when the nitrogen flow rate was 3.2 L/min, the injection rate of suspension was 200 μL/min, the concentration of suspension was 10% Me2SO, the receiving position was 6 cm from the nozzle and the needle type was 30G, the viability of HepG2 was the highest. The optimized spray parameters can be used as a reference for other small cell micro-droplet spray vitrification preservation.