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A glass micropipette vacuum technique of cerebrospinal fluid sampling in C57BL/6 mice

ZHANG Lu-Lin¹, YUAN Yu¹, LIANG Mei-Yu¹, LIU Ming-Xin¹, WANG Dong-Xia^1,2, XIE Jun-Xia¹, SONG Ning^1,*

¹Department of Physiology and Pathophysiology, School of Basic Medicine, Institute of Brain Science and Disease, Qingdao University, Qingdao 266071, China；²CT Center, Qingdao Jimo District Hospital of Traditional Chinese Medicine, Qingdao 266200, China

Abstract

The purpose of this study was to establish a suitable method for extracting cerebrospinal fluid (CSF) from C57BL/6 mice. A patch clamp electrode puller was used to draw a glass micropipette, and a brain stereotaxic device was used to fix the mouse’s head at an angle of 135° from the body. Under a stereoscopic microscope, the skin and muscle tissue on the back of the mouse’s head were separated, and the dura mater at the cerebellomedullary cistern was exposed. The glass micropipette (with an angle of 20° to 30° from the dura mater) was used to puncture at a point 1 mm inboard of Y-shaped dorsal vertebral artery for CSF sampling. After the first extraction, the glass micropipette was connected with a 1 mL sterile syringe to form a negative pressure device for the second extraction. The results showed that the successful rate of CSF extraction was 83.33% (30/36). Average CSF extraction amount was (7.16 ± 0.43) μL per mouse. In addition, C57BL/6 mice were given intranasally ferric ammonium citrate (FAC) to establish a model of brain iron accumulation, and the CSF extraction technique established in the present study was used for sampling. The results showed that iron content in the CSF from the normal saline control group was not detected, while the iron content in the CSF from FAC-treated group was (76.24 ± 38.53) μmol/L, and the difference was significant. These results suggest that glass micropipette vacuum technique of CSF sampling established in the present study has the advantages of simplicity, high success rate, large extraction volume, and low bleeding rate, and is suitable for the research on C57BL/6 mouse neurological disease models.

Key words: cerebrospinal fluid; C57BL/6 mice; ferric ammonium citrate

Received: 　　Accepted:

Corresponding author: 宋宁　　E-mail: ningsong@qdu.edu.cn

DOI: 10.13294/j.aps.2023.0025

Citing This Article：

ZHANG Lu-Lin, YUAN Yu, LIANG Mei-Yu, LIU Ming-Xin, WANG Dong-Xia, XIE Jun-Xia, SONG Ning. A glass micropipette vacuum technique of cerebrospinal fluid sampling in C57BL/6 mice. Acta Physiol Sin 2023; 75 (2): 197-204 (in Chinese with English abstract).