Approach for patch-clamping using an upright microscope with z-axis movable stage
Нема приказа
Аутори
Perić, MinaBataveljić, Danijela
Bijelić, Dunja
Milicević, Katarina
Andjus, Pavle R.
Pristov, Jelena Bogdanovic
Nikolić, Ljiljana
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 eas...ily 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.
Кључне речи:
patch-clamp / microscopy / life science / glial cells / cellular physiologyИзвор:
Microscopy Research and Technique, 2022, 85, 6, 2095-2104Издавач:
- Wiley, Hoboken
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200178 (Универзитет у Београду, Биолошки факултет) (RS-MESTD-inst-2020-200178)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200007 (Универзитет у Београду, Институт за биолошка истраживања 'Синиша Станковић') (RS-MESTD-inst-2020-200007)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200053 (Универзитет у Београду, Институт за мултидисциплинарна истраживања) (RS-MESTD-inst-2020-200053)
DOI: 10.1002/jemt.24066
ISSN: 1059-910X
PubMed: 35088507
WoS: 000747718200001
Scopus: 2-s2.0-85123799490
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Perić, Mina AU - Bataveljić, Danijela AU - Bijelić, Dunja AU - Milicević, Katarina AU - Andjus, Pavle R. AU - Pristov, Jelena Bogdanovic AU - Nikolić, Ljiljana PY - 2022 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1531 AB - 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. PB - Wiley, Hoboken T2 - Microscopy Research and Technique T1 - Approach for patch-clamping using an upright microscope with z-axis movable stage EP - 2104 IS - 6 SP - 2095 VL - 85 DO - 10.1002/jemt.24066 ER -
@article{ author = "Perić, Mina and Bataveljić, Danijela and Bijelić, Dunja and Milicević, Katarina and Andjus, Pavle R. and Pristov, Jelena Bogdanovic and Nikolić, Ljiljana", year = "2022", abstract = "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.", publisher = "Wiley, Hoboken", journal = "Microscopy Research and Technique", title = "Approach for patch-clamping using an upright microscope with z-axis movable stage", pages = "2104-2095", number = "6", volume = "85", doi = "10.1002/jemt.24066" }
Perić, M., Bataveljić, D., Bijelić, D., Milicević, K., Andjus, P. R., Pristov, J. B.,& Nikolić, L.. (2022). Approach for patch-clamping using an upright microscope with z-axis movable stage. in Microscopy Research and Technique Wiley, Hoboken., 85(6), 2095-2104. https://doi.org/10.1002/jemt.24066
Perić M, Bataveljić D, Bijelić D, Milicević K, Andjus PR, Pristov JB, Nikolić L. Approach for patch-clamping using an upright microscope with z-axis movable stage. in Microscopy Research and Technique. 2022;85(6):2095-2104. doi:10.1002/jemt.24066 .
Perić, Mina, Bataveljić, Danijela, Bijelić, Dunja, Milicević, Katarina, Andjus, Pavle R., Pristov, Jelena Bogdanovic, Nikolić, Ljiljana, "Approach for patch-clamping using an upright microscope with z-axis movable stage" in Microscopy Research and Technique, 85, no. 6 (2022):2095-2104, https://doi.org/10.1002/jemt.24066 . .