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Cell culture confluency estimation, transfection efficiency and total cell counting applications on the InCellis ® Smart Cell Imaging System
Author(s) -
Antanaviciute Laima,
Dubacq Sophie,
Varet Olivier
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.lb585
Subject(s) - cell culture , cell counting , cell growth , cell , hela , cell cycle , biology , genetics
Cell biology studies of cell proliferation, invasive behaviour of cells, and cell line expression require a rapid and robust cell line confluency estimation, transfection efficiency investigation, and accurate total cell counting. Accuracy and efficiency are extremely important in cell culture quality check applications. Incorrect observations might lead not only to inaccurate results but also to incorrect conclusions and recommendations. In this study, breast cancer cell line (MCF‐7) was used to accurately estimate cell line confluency and total cell counting in an automated manner using embedded applications on the InCellis ® Smart Cell Imaging System (Bertin Technologies). A series of images of the cell line were taken in phase contrast mode using 10× and 20× objectives on day‐1, day‐2 and day‐3 at the same time in order to check the accuracy of cell confluency. The cell line confluency ranged from 12% – 76% on day‐1 through day‐3 with the 10× objective, and ranged from 14% to 77% with the 20× objective respectively. Furthermore, the results obtained from the cell counting application embedded on the InCellis ® were compared to the manual approach performed with Malassez cell counting method that was treated as a control across the 3‐day experiment. The results demonstrated that both methods are correlated and showed an increase of the total cell number as expected (40K on day‐1 ‐ 270K on day‐3). To validate the automated transfection efficiency application, Hela cell culture lines and GFP label were used. The 3‐step process allowed easy and spontaneous observation of the percentage of transfected cells and the overall quality inspection of the cell line. The confluency and cell counting applications supply robust results with a stain‐free method in comparison to the standard method. The transfection application provided a rapid and user‐friendly method for estimation of transfected cells before use in further analysis. The automated applications not only provide consistent results but also significantly reduce the hands‐on time for all cell based assays. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .