Abstract
Laser interference microscopy (LIM) is a promising label-free method for single-cell research applicable to cell viability assessment in the studies of mammalian cells. This paper describes the development of a sensitive and reproducible method for assessing cell viability using LIM. The method, based on associated signal processing techniques, has been developed as a result of real-time investigation in phase thickness fluctuations of viable and non-viable MCF-7 cells, reflecting the presence and absence of their metabolic activity. As evinced by the values of the variable vc, this variable determines the viability of a cell only in the attached state (vc exceeds 20 nm2 for viable attached cells). The critical value of the power spectrum slope βc of the phase thickness fluctuations equals 1.00 for attached MCF-7 cells and 0.71 for suspended cells. The slope of the phase fluctuations' power spectrum for MCF-7 cells was determined to exceed the threshold value of βc for a living cell, otherwise the cell is dead. The results evince the power spectrum slope as the most appropriate indicator of cell viability, while the integrated evaluation criterion (vc and βc values) can be used to assay the viability of attached cells.
Highlights
MCF-7 cells are commonly used adherent breast cancer cells isolated from the pleural effusion of a 69-year-old woman with metastatic disease in 1973 by Dr Soule and colleagues at the Michigan Cancer Foundation [38,39]
MCF-7 cells have been used as a model in cell
To monitor the viability of cells in the suspended and attached states in real-time and by a non-invasive mode, we described a new label-free Laser interference microscopy (LIM)-based method that can be multiplexed with other cell assays
Summary
The assessment of viability plays an important role in most mammalian cell-based biological, biomedical, and pharmacological studies [1]. In this case, viability means the physiological state of cells that allows carrying out their regular functions and ensuring tissue-specific activities and the possibility of cell division [2]. Most extant methods for cell viability determination rely on assessing the morphological integrity of cells or their proliferative or metabolic activities [3,4]. To determine the number of viable cells, detection/visualization methods, such as bright-field and fluorescence microscopy, flow cytometry, and spectrophotometry by a plate reader, have often been used
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