BioTek July 2019 - 21

Paul Held, J.C., Peter Banks, High Contrast Brightfield:
Enabling microplate-based automated label-free cell
counting BioTek Resources, 2016.

Joe Clayton, P.B., A Guide to Label-free Cell Counting
using High Contrast Brightfield. BioTek Resources 2016.

Tacar, O., P. Sriamornsak, and C.R. Dass, Doxorubicin:
an update on anticancer molecular action, toxicity and
novel drug delivery systems. J Pharm Pharmacol, 2013.
65(2): p. 157-70.

Fornari, F.A., et al., Interference by doxorubicin with
DNA unwinding in MCF-7 breast tumor cells. Mol
Pharmacol, 1994. 45(4): p. 649-56.

Konicek, B.W., et al., Therapeutic inhibition of MAP
kinase interacting kinase blocks eukaryotic initiation
factor 4E phosphorylation and suppresses outgrowth of
experimental lung metastases. Cancer Res, 2011. 71(5):
p. 1849-57.

Altman, J.K., et al., Inhibition of Mnk kinase activity
by cercosporamide and suppressive effects on acute
myeloid leukemia precursors. Blood, 2013. 121(18): p.
3675- 81.

Figure 8. Qualitative analysis of kinetic cell proliferation provides valuable
insight into phenotypic response to drug treatment. NIH3T3 proliferation
images 36 hours after treatment with indicated concentration of doxorubicin. At
10 nM doxorubicin cell division is inhibited without causing overt cytotoxicity.
At 100 nM and higher concentrations signs of cytotoxicity are evident.

Conclusions
Coupling the cell-handling abilities of the BioSpa™
8 with the imaging capabilities of the Cytation™ 5
provides a fully automated system to conduct accurate and reproducible long-term proliferation studies. High contrast brightfield cell counting enables
quantitative analysis of cell growth without the need
for disruptive labels. In addition to quantitative measures, this system can be used to conduct qualitative
analysis of cell phenotypes over time to further characterize drug treatment response or conduct targeted gene disruption studies. Proliferation assays can
be run in 96- or 384-well microplates for mediumto high-throughput screening. Together, the BioSpa
8 and Cytation 5, along with the powerful Gen™ 5
image analysis tools, provide an elegant and robust
solution for a broad range of kinetic cell proliferation
applications.

About the Author
Joe Clayton, Ph.D., is Principal Scientist, BioTek
Instruments, Inc.

References
1.

Drey, L.L., M.C. Graber, and J. Bieschke, Counting
unstained, confluent cells by modified bright-field
microscopy. Biotechniques, 2013. 55(1): p. 28-33.



Movie 1: NIH3T3 treated with

Movie 2: NIH3T3 treated with vehicle

0.1 µM doxorubicin

https://www.biotek.com/videos/Applications/Movie%201%20NIH%203T3%200.1%20uM%20doxorubicin%20Proliferation%20Movie%20high%20contrast%20brightfield.mp4 https://www.biotek.com/videos/Applications/Movie%202%20NIH%203T3%20Proliferation%20Movie%20high%20contrast%20brightfield.mp4

BioTek July 2019

Table of Contents for the Digital Edition of BioTek July 2019