Commentary in 2014

Developments in electrical and optical recording technology are scaling up the size of neuronal populations that can be monitored simultaneously. Light-sheet imaging is rapidly gaining traction as a method for optically interrogating activity in large networks and presents both opportunities and challenges for understanding circuit function.

In light sheet–based fluorescence microscopy (LSFM), optical sectioning in the excitation process minimizes fluorophore bleaching and phototoxic effects. Because biological specimens survive long-term three-dimensional imaging at high spatiotemporal resolution, LSFM has become the tool of choice in developmental biology.

Ten years of development in light-sheet microscopy have led to spectacular demonstrations of its capabilities. The technology is ready to assist biologists in tackling scientific problems, but are biologists ready for it? Here we discuss the interdisciplinary challenges light-sheet microscopy presents for biologists and highlight available resources.

The ability to convert somatic cells to induced pluripotent stem cells has immense potential to further our understanding of development and disease mechanisms, and for cellular therapy. Before researchers can achieve these goals, they must expand current methodology to incorporate animal models and quantitative descriptions of the essential phenomena driving reprogramming.

A decade of advances in genome engineering technologies has enabled the editing of genome sequences much like one edits computer code; many more applications for precisely manipulating genome structure and function are on the horizon.

Our choice, among many candidates, of the ten areas of methods development with the most impact on biological research over the last decade. Visit Methagora to browse Nature Methods papers in some of these areas.

A revolution in DNA sequencing technology has enabled new insights from thousands of genomes sequenced across taxa.

Much of our knowledge about biological systems has been obtained by examining ensembles of molecules. However, this has begun to change because of the unprecedented precision and clarity afforded by single-molecule measurements. The last decade has seen amazing advances in the resolution and complexity of these methods, making it possible to ask and answer entirely new types of biological questions.

The optogenetic revolution is transforming neuroscience. The dramatic recent progress in using light to both control and read out neural activity has highlighted the need for better probes, improved light delivery and more careful interpretation of results, which will all be required for optogenetics to fully realize its remarkable potential.

We argue that standard thermodynamic considerations and scaling laws show that a single cell cannot substantially raise its temperature by endogenous thermogenesis. This statement seriously questions the interpretations of recent work reporting temperature heterogeneities measured in single living cells.

'Irreproducibility' is symptomatic of a broader challenge in measurement in biomedical research. From the US National Institute of Standards and Technology (NIST) perspective of rigorous metrology, reproducibility is only one aspect of establishing confidence in measurements. Appropriate controls, reference materials, statistics and informatics are required for a robust measurement process. Research is required to establish these tools for biological measurements, which will lead to greater confidence in research results.

Authors discuss how synthetic biology approaches could be applied to assemble synthetic quasibiological systems able to replicate and evolve, illuminating universal properties of life and the search for its origins.

Single-molecule super-resolution techniques emerged only several years ago but have revolutionized fluorescence microscopy of cellular structures. We discuss some key principles of these techniques, point out pitfalls, highlight recent developments and identify opportunities for the future.