A randomised approach for enumerating migrated cells in a transwell migration assay

The transwell migration assay is commonly used for assessing cell migration. It involves the enumeration of cells that have migrated across a pore-containing membrane. We describe a randomised approach to quantifying migrated cells and compare it to a conventional full cell count. We used ATP as a c...

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Main Authors: Kanesan, Livashini, Omar Zaki, Siti Sarah, Vidyadaran, Sharmili
Format: Article
Language:English
Published: Faculty of Medicine and Health Sciences, Universiti Putra Malaysia 2018
Online Access:http://psasir.upm.edu.my/id/eprint/66176/
http://psasir.upm.edu.my/id/eprint/66176/1/2018121312121808_MJMHS_December_2018.pdf
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author Kanesan, Livashini
Omar Zaki, Siti Sarah
Vidyadaran, Sharmili
author_facet Kanesan, Livashini
Omar Zaki, Siti Sarah
Vidyadaran, Sharmili
author_sort Kanesan, Livashini
building UPM Institutional Repository
collection Online Access
description The transwell migration assay is commonly used for assessing cell migration. It involves the enumeration of cells that have migrated across a pore-containing membrane. We describe a randomised approach to quantifying migrated cells and compare it to a conventional full cell count. We used ATP as a chemoattractant and automatic cell quantification performed on all fields (Full count; FC) or 10 randomly selected fields (Randomised count; RC). The two methods were compared by evaluating standard deviations (SD), coefficient of variation (CV) and using the Bland-Altman analysis. The dispersion of data is higher with the RC approach (3.77-6.66% CV for control; 3.89-4.48% CV for ATP-treated wells) compared to FC (0.27-0.46% CV for control; 0.05-0.09% CV for ATP-treated wells), but are acceptable considering that the number of migrated cells are in the thousands. Both methods verified that an ATP migration assay for BV2 microglia was established, demonstrating that the RC approach is reliable and comparable to a full count.
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spelling upm-661762019-02-11T03:59:47Z http://psasir.upm.edu.my/id/eprint/66176/ A randomised approach for enumerating migrated cells in a transwell migration assay Kanesan, Livashini Omar Zaki, Siti Sarah Vidyadaran, Sharmili The transwell migration assay is commonly used for assessing cell migration. It involves the enumeration of cells that have migrated across a pore-containing membrane. We describe a randomised approach to quantifying migrated cells and compare it to a conventional full cell count. We used ATP as a chemoattractant and automatic cell quantification performed on all fields (Full count; FC) or 10 randomly selected fields (Randomised count; RC). The two methods were compared by evaluating standard deviations (SD), coefficient of variation (CV) and using the Bland-Altman analysis. The dispersion of data is higher with the RC approach (3.77-6.66% CV for control; 3.89-4.48% CV for ATP-treated wells) compared to FC (0.27-0.46% CV for control; 0.05-0.09% CV for ATP-treated wells), but are acceptable considering that the number of migrated cells are in the thousands. Both methods verified that an ATP migration assay for BV2 microglia was established, demonstrating that the RC approach is reliable and comparable to a full count. Faculty of Medicine and Health Sciences, Universiti Putra Malaysia 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/66176/1/2018121312121808_MJMHS_December_2018.pdf Kanesan, Livashini and Omar Zaki, Siti Sarah and Vidyadaran, Sharmili (2018) A randomised approach for enumerating migrated cells in a transwell migration assay. Malaysian Journal of Medicine and Health Sciences, 14 (SP3). pp. 50-53. ISSN 1675-8544; ESSN: 2636-9346 http://www.medic.upm.edu.my/upload/dokumen/2018121312121808_MJMHS_December_2018.pdf
spellingShingle Kanesan, Livashini
Omar Zaki, Siti Sarah
Vidyadaran, Sharmili
A randomised approach for enumerating migrated cells in a transwell migration assay
title A randomised approach for enumerating migrated cells in a transwell migration assay
title_full A randomised approach for enumerating migrated cells in a transwell migration assay
title_fullStr A randomised approach for enumerating migrated cells in a transwell migration assay
title_full_unstemmed A randomised approach for enumerating migrated cells in a transwell migration assay
title_short A randomised approach for enumerating migrated cells in a transwell migration assay
title_sort randomised approach for enumerating migrated cells in a transwell migration assay
url http://psasir.upm.edu.my/id/eprint/66176/
http://psasir.upm.edu.my/id/eprint/66176/
http://psasir.upm.edu.my/id/eprint/66176/1/2018121312121808_MJMHS_December_2018.pdf