Using bespoke fluorescence microscopy to study the soft condensed matter of living cells at the single molecule level
Journal of Physics: Conference Series 286:1 (2011)
Abstract:
The use of bespoke imaging tools and analysis can offer significant insight into the living counterpart of soft condensed matter. The soft matter of biological systems consists of molecular building blocks, a staple of which is protein. Protein molecules, so small that 1 billion would fit on the full-stop at the end of this sentence, carry out most of the vital activities in living cells. Many of these processes require the assembly of multiple proteins into remarkable biological machines. Obtaining the blueprints for the architecture of these machines is essential for understanding the workings of the cell. Here, we discuss recent biological physics experiments on functional single-celled organisms in which one can apply bespoke fluorescence microscopy imaging and analysis to monitor the number and dynamics of several different proteins at the nanometre length scale to a precision of single molecules. © Published under licence by IOP Publishing Ltd 2011.Functioning nanomachines seen in real-time in living bacteria using single-molecule and super-resolution fluorescence imaging.
Int J Mol Sci 12:4 (2011) 2518-2542
Abstract:
Molecular machines are examples of "pre-established" nanotechnology, driving the basic biochemistry of living cells. They encompass an enormous range of function, including fuel generation for chemical processes, transport of molecular components within the cell, cellular mobility, signal transduction and the replication of the genetic code, amongst many others. Much of our understanding of such nanometer length scale machines has come from in vitro studies performed in isolated, artificial conditions. Researchers are now tackling the challenges of studying nanomachines in their native environments. In this review, we outline recent in vivo investigations on nanomachines in model bacterial systems using state-of-the-art genetics technology combined with cutting-edge single-molecule and super-resolution fluorescence microscopy. We conclude that single-molecule and super-resolution fluorescence imaging provide powerful tools for the biochemical, structural and functional characterization of biological nanomachines. The integrative spatial, temporal, and single-molecule data obtained simultaneously from fluorescence imaging open an avenue for systems-level single-molecule cellular biophysics and in vivo biochemistry.Using bespoke fluorescence microscopy to study the soft condensed matter of living cells at the single molecule level
CONDENSED MATTER AND MATERIALS PHYSICS CONFERENCE (CMMP10) 286 (2011) ARTN 012001
Shining the spotlight on functional molecular complexes: The new science of single-molecule cell biology.
Commun Integr Biol 3:5 (2010) 415-418
Abstract:
Single-molecule research is emerging as one of the fastest growing fields within the biosciences. Historically, most of the techniques employed have operated largely in the world of the test tube in which the components of the biological system under investigation have been extracted and purified from cells to reduce them to just the key ingredients under study, and this research has involved novel, pioneering methods of biophysics to obtain single-molecule measurements. What has emerged recently is the technical ability to now perform key single-molecule experiments whilst retaining the native biological context-namely to do single-molecule experiments on functional living cells. This presents essentially a new science of "single-molecule cell biology", which combines classical cell biology approaches with modern single-molecule biophysics. Here, key recent studies which have pushed back the boundaries of this field are discussed.A general approach for segmenting elongated and stubby biological objects: Extending a chord length transform with the radon transform
2010 7th IEEE International Symposium on Biomedical Imaging: From Nano to Macro, ISBI 2010 - Proceedings (2010) 161-164