Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines

Article

Busschots, S., O'Toole, S., O'Leary, J. and Stordal, B. 2015. Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines. MethodsX. 2, pp. 8-13. https://doi.org/10.1016/j.mex.2014.11.002
TypeArticle
TitleNon-invasive and non-destructive measurements of confluence in cultured adherent cell lines
AuthorsBusschots, S., O'Toole, S., O'Leary, J. and Stordal, B.
Abstract

Many protocols used for measuring the growth of adherent monolayer cells in vitro are invasive, destructive and do not allow for the continued, undisturbed growth of cells within flasks. Protocols often use indirect methods for measuring proliferation. Microscopy techniques can analyse cell proliferation in a non-invasive or non-destructive manner but often use expensive equipment and software algorithms. In this method images of cells within flasks are captured by photographing under a standard inverted phase contract light microscope using a digital camera with a camera lens adaptor. Images are analysed for confluence using ImageJ freeware resulting in a measure of confluence known as an Area Fraction (AF) output. An example of the AF method in use on OVCAR8 and UPN251 cell lines is included.
•Measurements of confluence from growing adherent cell lines in cell culture flasks is obtained in a non-invasive, non-destructive, label-free manner.
•The technique is quick, affordable and eliminates sample manipulation.
•The technique provides an objective, consistent measure of when cells reach confluence and is highly correlated to manual counting with a haemocytometer. The average correlation co-efficient from a Spearman correlation (n = 3) was 0.99 ± 0.008 for OVCAR8 (p = 0.01) and 0.99 ± 0.01 for UPN251 (p = 0.01) cell lines.

KeywordsConfluence; Cell count; Phase-contrast microscopy; ImageJ; Area fraction; Non-destructive
Research GroupBiomarkers for Cancer group
LanguageEnglish
PublisherElsevier
JournalMethodsX
ISSN
Electronic2215-0161
Publication dates
Online25 Nov 2014
Print2015
Publication process dates
Deposited08 Jan 2016
Accepted14 Nov 2014
Output statusPublished
Publisher's version
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Copyright Statement

© 2015 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/3.0/).

Web address (URL)https://www.sciencedirect.com/science/article/pii/S2215016114000260
Digital Object Identifier (DOI)https://doi.org/10.1016/j.mex.2014.11.002
Web of Science identifierWOS:000370471500002
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