Live Cell Imaging of the Cytoskeleton

Author(s):  
Eve G. Stringham ◽  
Nancy Marcus-Gueret ◽  
Laura Ramsay ◽  
Kristopher L. Schmidt
2019 ◽  
Vol 91 (15) ◽  
pp. 10095-10101 ◽  
Author(s):  
Palanisamy Ravichandiran ◽  
Sivakumar Allur Subramaniyan ◽  
Antony Paulraj Bella ◽  
Princy Merlin Johnson ◽  
Ae Rhan Kim ◽  
...  

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.


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