Abstract
One of the main aspects investigated in potential therapeutic compounds is their effect on cells viability and proliferative ability. Although various methods have been developed to investigate these aspects, these methods present with shortcomings in terms of either cost, availability, accuracy, precision, or throughput. This study describes a simple, economic, reproducible, and high-throughput assay to quantify cell death and proliferation. In this assay, adherent cells are fixed, stained with trypan blue, and measured for trypan blue internalization using a spectrophotometric absorbance plate reader. Corresponding cell counts to the absorbance measurements are extrapolated from a standard curve. This assay was used to measure the effect of dimethyl sulfoxide (DMSO) on the viability of breast and lung cancer cells. Decrease in cell count associated with the increase in DMSO percentage and exposure time. The assay’s results closely correlated with the conventional trypan blue exclusion assay (Pearson correlation coefficient (r) > 0.99; p < 0.0001), but with higher precision. The assay developed in this study can be used for various applications such as optimization, cell treatment investigations, proliferation, and cytotoxicity studies.
Highlights
Cell counting is a common, important practice in cell culture used on daily basis for different applications
Various alternative approaches were developed aiming at fulfilling these requirements and overcoming original methods’ shortcomings. These approaches were based on various concepts including automation, flow cytometry, impedance, tetrazolium salts, methylene blue, microscopy, fluorescence, and protein quantification
Various cell-counting approaches were developed to fulfil application-specific needs, these approaches are either low throughput or relatively cost-demanding
Summary
Cell counting is a common, important practice in cell culture used on daily basis for different applications. Various alternative approaches were developed aiming at fulfilling these requirements and overcoming original methods’ shortcomings. The use of haemocytometers and counting chambers was the standardized cell-counting approach Due to their various limitations (i.e., low accuracy, low precision, low throughput, high probability of variations, and susceptibility to various human error sources), the use of these methods became limited to simple applications (Ongena et al 2010; Tucker et al 1994). An improved, automated modification of these methods was developed aiming at increasing the cell-count throughput It failed to overcome the other shortcomings due to the similarities in sample preparation and mounting onto the special counting slides/chambers (Camacho-Fernandez et al 2018). Despite the various adjustment options, automated counters remain prone to fail in realizing varying cellular shapes and sizes pooled in the same sample as well as outof-focus cells in different focal planes (Camacho-Fernandez et al 2018)
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
