Analysis of microglial migration by a micropipette assay.
Microglial cells have important roles in maintaining brain homeostasis, and they are implicated in multiple brain diseases. There is currently interest in investigating microglial migration that results in cell accumulation at focal sites of injury. Here we describe a protocol for rapidly triggering and monitoring microglial migration by using a micropipette assay. This protocol is an adaptation of the axon turning assay using microglial cells. Chemoattractants released from the micropipette tip produce a chemotactic gradient that induces robust microglial migration. In combination with microscopic imaging, this assay allows simultaneous recording of cell movement and subcellular compartment trafficking, along with quantitative analysis. The actual handling time for the assay takes ∼2-3 h in total. The protocol is simple, inexpensive and convenient to set up, and it can be adopted to examine cell migration in multiple cell types, including cancer cells with a wide range of chemical signals.
1] Britton Chance Center for Biomedical Photonics, The Key Laboratory of Biomedical Photonics of The Ministry of Education, Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, China. [2] Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Hang-jun
HJ
Wu
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Yi-jun
YJ
Liu
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Hui-quan
HQ
Li
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Cong
C
Chen
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Ying
Y
Dou
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Hui-fang
HF
Lou
Department of Neurobiology and Anatomy, School of Medicine, Tongji University, Shanghai, China.
Margaret S
MS
Ho
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Xiao-ming
XM
Li
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Zhihua
Z
Gao
Department of Neurobiology, The Key Laboratory of Medical Neurobiology, The Ministry of Health of China, Zhejiang Provincial Key Laboratory of Neurobiology, Zhejiang University School of Medicine, Hangzhou, China.
Shumin
S
Duan
D000255:Adenosine Triphosphate / Q000378:metabolism; D001921:Brain / Q000166:cytology*; D001927:Brain Diseases / Q000276:immunology*; D054443:Cell Migration Assays / Q000379:methods*; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D002630:Chemotactic Factors / Q000494:pharmacology; D006801:Humans; D017628:Microglia / Q000276:immunology / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*
D002630:Chemotactic Factors; D000255:Adenosine Triphosphate