The influence of microinjection parameters on cell survival and procedure efficiency
Microinjection is a method commonly used to deliver various substances into cells. The procedure is performed on a widefield microscope stage using fine glass needle to penetrate the cell membrane. Microinjection can be carried out using a manual or semi-automatic mode. For commercially available eq...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016123001097 |
| _version_ | 1797796489503703040 |
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| author | Joanna Hajduk Konrad Szajna Bartosz Lisowski Zenon Rajfur |
| author_facet | Joanna Hajduk Konrad Szajna Bartosz Lisowski Zenon Rajfur |
| author_sort | Joanna Hajduk |
| collection | DOAJ |
| description | Microinjection is a method commonly used to deliver various substances into cells. The procedure is performed on a widefield microscope stage using fine glass needle to penetrate the cell membrane. Microinjection can be carried out using a manual or semi-automatic mode. For commercially available equipment currently reported microinjection success rate and cell viability are relatively low (around 50% for both indicators). Here, for the first time, we systematically show how the microinjection effectiveness and cell viability are influenced by needle diameter and chosen microinjection mode. We found that manual mode entailed a higher injection rate, reducing cell viability at the same time. The reduction in needle diameter caused a significant increase in cell survival rate (from 43 to 73% for manual mode and from 58% to 86% for semi-automatic mode) and did not affect significantly the success rate. Our findings will help optimize this method in the context of cell biology research. • This study shows how to improve microinjection parameters, such as procedure efficiency and cell survival rate, for commercially available equipment. • Manual mode, in comparison with semi-automatic mode, results in higher microinjection efficiency, but lower cell survival rate. • The increase in micropipette diameter causes lower cell viability and a higher microinjection success rate. |
| first_indexed | 2024-03-13T03:33:52Z |
| format | Article |
| id | doaj.art-e12625f5f9404a4ba50db78f3067e02c |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-03-13T03:33:52Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-e12625f5f9404a4ba50db78f3067e02c2023-06-24T05:17:22ZengElsevierMethodsX2215-01612023-01-0110102107The influence of microinjection parameters on cell survival and procedure efficiencyJoanna Hajduk0Konrad Szajna1Bartosz Lisowski2Zenon Rajfur3Doctoral School of Exact and Natural Sciences, Jagiellonian University, Łojasiewicza 11, Krakow 30-348, Poland; Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Łojasiewicza 11, Krakow 30-348, PolandFaculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Łojasiewicza 11, Krakow 30-348, PolandFaculty of Pharmacy, Jagiellonian University Medical College, Medyczna 9, Kraków 30-688, Poland; Corresponding authors.Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Łojasiewicza 11, Krakow 30-348, Poland; Jagiellonian Center of Biomedical Imaging, Jagiellonian University, Kraków 30-348, Poland; Corresponding author at: Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, Łojasiewicza 11, Krakow 30-348, PolandMicroinjection is a method commonly used to deliver various substances into cells. The procedure is performed on a widefield microscope stage using fine glass needle to penetrate the cell membrane. Microinjection can be carried out using a manual or semi-automatic mode. For commercially available equipment currently reported microinjection success rate and cell viability are relatively low (around 50% for both indicators). Here, for the first time, we systematically show how the microinjection effectiveness and cell viability are influenced by needle diameter and chosen microinjection mode. We found that manual mode entailed a higher injection rate, reducing cell viability at the same time. The reduction in needle diameter caused a significant increase in cell survival rate (from 43 to 73% for manual mode and from 58% to 86% for semi-automatic mode) and did not affect significantly the success rate. Our findings will help optimize this method in the context of cell biology research. • This study shows how to improve microinjection parameters, such as procedure efficiency and cell survival rate, for commercially available equipment. • Manual mode, in comparison with semi-automatic mode, results in higher microinjection efficiency, but lower cell survival rate. • The increase in micropipette diameter causes lower cell viability and a higher microinjection success rate.http://www.sciencedirect.com/science/article/pii/S2215016123001097Adherent cells manual and semi-automatic microinjection |
| spellingShingle | Joanna Hajduk Konrad Szajna Bartosz Lisowski Zenon Rajfur The influence of microinjection parameters on cell survival and procedure efficiency MethodsX Adherent cells manual and semi-automatic microinjection |
| title | The influence of microinjection parameters on cell survival and procedure efficiency |
| title_full | The influence of microinjection parameters on cell survival and procedure efficiency |
| title_fullStr | The influence of microinjection parameters on cell survival and procedure efficiency |
| title_full_unstemmed | The influence of microinjection parameters on cell survival and procedure efficiency |
| title_short | The influence of microinjection parameters on cell survival and procedure efficiency |
| title_sort | influence of microinjection parameters on cell survival and procedure efficiency |
| topic | Adherent cells manual and semi-automatic microinjection |
| url | http://www.sciencedirect.com/science/article/pii/S2215016123001097 |
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