PMID- 34799183 OWN - NLM STAT- MEDLINE VI - 40 IP - 6 TI - Microscopy imaging of living cells in metabolic engineering. PG - 752-765 CI - Copyright © 2021 Elsevier Ltd. All rights reserved. LA - eng PT - Journal Article PT - Review PT - Research Support, Non-U.S. Gov't PL - England TA - Trends Biotechnol JT - Trends in biotechnology JID - 8310903 IS - 1879-3096 (Electronic) LID - S0167-7799(21)00256-0 [pii] LID - 10.1016/j.tibtech.2021.10.010 [doi] FAU - Lv, Xueqin AU - Lv X AD - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, China. FAU - Jin, Ke AU - Jin K AD - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, China. FAU - Sun, Guoyun AU - Sun G AD - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, China. FAU - Ledesma-Amaro, Rodrigo AU - Ledesma-Amaro R AD - Department of Bioengineering and Imperial College Centre for Synthetic Biology, Imperial College London, London SW72AZ, UK. FAU - Liu, Long AU - Liu L AD - Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China; Science Center for Future Foods, Jiangnan University, Wuxi 214122, China. Electronic address: longliu@jiangnan.edu.cn. IS - 0167-7799 (Linking) SB - IM MH - *Metabolic Engineering MH - Microscopy, Confocal MH - Microscopy, Fluorescence/methods OTO - NOTNLM OT - *fluorescence microscopy OT - *live-cell imaging OT - *metabolic engineering OT - *super-resolution microscopy OT - *synthetic biology LR - 20220718 DP - 202206 DEP - 20211116 AB - Microscopy imaging of living cells is becoming a pivotal, noninvasive, and highly specific tool in metabolic engineering to visualize molecular dynamics in industrial microorganisms. This review describes the different microscopy methods, from fluorescence to super resolution, with application in microbial bioengineering. Firstly, the role and importance of microscopy imaging is analyzed in the context of strain design. Then, the advantages and disadvantages of different microscopy technologies are discussed, including confocal laser scanning microscopy (CLSM), spatial light interference microscopy (SLIM), and super-resolution microscopy, followed by their applications in synthetic biology. Finally, the future perspectives of live-cell imaging and their potential to transform microbial systems are analyzed. This review provides theoretical guidance and highlights the importance of microscopy in understanding and engineering microbial metabolism.