Approach for patch-clamping using an upright microscope with z-axis movable stage.
2022
Authors:
Perić, MinaBataveljić, Danijela
Bijelić, Dunja
Milićević, Katarina
Andjus, Pavle R.
Bogdanović Pristov, Jelena
Nikolić, Ljiljana
Document Type:
Article (Published version)
,
© 2022 Wiley Periodicals LLC.
Metadata
Show full item recordAbstract:
We describe an approach for studying the physiology of single live cells using the conceptionally novel upright microscope/patch-clamp configuration. Electrophysiology experiments typically require a microscope with the fixed stage position and the motion control of the microscope objective. Here, we demonstrate that a microscope with a z-axis movable stage and a fixed objective can also be efficiently used in combination with the patch-clamp technique. We define a set of underlying principles governing the operation of this microscope/patch-clamp configuration and demonstrate its performance in practice using cultured astrocytes, microglia, and oligodendrocytes. Experimental results show that our custom configuration provides stable recordings, has a high success rate of the whole-cell patch-clamp trials, can be effectively applied to study cellular physiology of glial cells, and provides comparable performance and usability to the commercially available systems. Our system can be easily replicated or adapted to suit the needs of the research groups and can be cost-effective in reducing the investments in purchasing additional equipment. We provide step-by-step instructions on implementing an upright microscope with z-axis movable stage as a routine workhorse for patch-clamping.
Keywords:
Cellular physiology; Glial cells; life science; Microscopy; Patch-clampSource:
Microscopy Research and Technique, 2022, 85, 6, 2095-2104Funding / projects:
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200007 (University of Belgrade, Institute for Biological Research 'Siniša Stanković') (RS-MESTD-inst-2020-200007)
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200178 (University of Belgrade, Faculty of Biology) (RS-MESTD-inst-2020-200178)
- Ministry of Science, Technological Development and Innovation of the Republic of Serbia, institutional funding - 200053 (University of Belgrade, Institute for Multidisciplinary Research) (RS-MESTD-inst-2020-200053)
DOI: 10.1002/jemt.24066
ISSN: 1059-910X
PubMed: 35088507
WoS: 000747718200001
Scopus: 2-s2.0-85123799490
URI
https://onlinelibrary.wiley.com/doi/10.1002/jemt.24066http://www.ncbi.nlm.nih.gov/pubmed/35088507
http://radar.ibiss.bg.ac.rs/handle/123456789/4790