A bioinformatics approach for the interrogation of molecular events in single cells: transforming fluorescent time-lapse microscopy images into numbers
https://doi.org/10.1186/1752-0509-1-S1-P31
© Khan et al; licensee BioMed Central Ltd. 2007
Published: 8 May 2007
Keywords
Exploring and exploiting the enormous potential for pharmacological modulation of the mammalian cell cycle are key goals for basic research and drug discovery. We have developed a critical advance – the high temporal resolution monitoring of cell cycle progression enabling the tracking of single cell checkpoint transitions in a non-invasive manner even within heterogeneous populations. The green fluorescent protein (GFP)-based probe has expression, location and destruction characteristics that shadow cyclin B1 dynamics in living cells [1]. The non-perturbing stealth reporter performance has been validated on high content to high throughput detection platforms comprising multi-well high-throughput screen (HTS) imaging, single cell kinetic-tracking and multi-parameter flow cytometry [1, 2]. Cyclin B1-GFP tracking provides sub-phase information on cell cycle progression, cell-cycle regulator dynamics in parallel with morphological landmarks and DNA content analysis. We have developed a bioinformatics software – FluroTRAK that semi automatically tracks the continuous progression of cell cycle traverse and encodes molecular readouts in bifurcating lineages. Single cell lineages which underlie the basic concept provide an elegant assay for determining the evolving and complex interplay for tumour survival at the single cell level. Our primary premise is that a bioinformatics approach dedicated to cell-based measurements provides an essential route to determining inter-event relationships revealing novel cellular and molecular event patterns.
Therefore our overall aim is to develop an integrated bioinformatics environment which encompasses the analysis tools to encode microscopy images into numbers and deposits the encoded data into relational databases. The intention is to provide a web-based interface with access to a suite of databases called CyMART http://biodiversity.cs.cf.ac.uk/cymart/ these provide databases enable robust hypothesis-driven data-mining and drug signature queries.
Conclusion
A typical encoded cell lineage. A cell lineage encoded from the progenitor cell (B), where the cell divides into two daughter cells 5 hours after the start of the experiment. The north daughter (BN) again divides at 28 hours into two daughter cells BNN and BNS while the south daughter BS divides at 44.66 hours into two daughter cells BSN and BSS. Four living cells (BNN,BNS, BSN and BSS) at the end of the experiment yielded four tracks labelled as track 1 2 3 and 4 respectively. Molecular readouts for each track encoded by the software and novel data format provides intra and inter lineage data relationship.
Authors’ Affiliations
References
- Thomas N, Goodyear ID: Stealth sensors: real-time monitoring of the cell cycle. Targets. 2003, 2: 26-23. 10.1016/S1477-3627(02)02282-1.View ArticleGoogle Scholar
- Thomas N: Lighting the circle of life. Cell Cycle. 2003, 2 (6): 545-549.PubMedView ArticleGoogle Scholar
Copyright
This article is published under license to BioMed Central Ltd.