Nature Method has recently sorted out the most noteworthy technologies in 2016, including protein labeling in cells, unraveling nuclear architecture, protein structure though time, precision optogenetics, highly multiplexed imaging, deep learning, subcellular maps and integrated single-cell profiles.

 
Florescent dyes are relatively small, having good photophysical properties and spectral span. Those features make fluorescent dyes attractive, and they are expected to replace fluorescent protein for protein labeling. Researchers are actively developing tools to label protein by dyes in living cells.

 
For most applications, fluorescent dye need to be able to achieve specific markers. There are already a number of tools that can do this, such as SNAP and Halo label, FlAsH and ReAsh and hexahistidine. Another method is the incorporation of unnatural amino acids in the protein translation process. Those non-natural amino acids themselves are fluorescent, or can be fluorescent through click chemistry.

 
Although these methods are increasingly popular, they also encountered some problems. For example, fluorescent dye has limited use in a multiple imaging. The development of such dyes is currently a very active research area. There is no doubt that in the future, the efficiency of the fluorescent dye marker will be greatly enhanced, which will further improve the ability of quantitative imaging.

 
Specific protein marker facilitates ultra-high resolution imaging in fixed cells and living cells. When the resolution is near tens of nanometers, the questions caused by labeling would stand out. For example, labeling with antibody will increase the target structure by about 10nm, while Secondary Antibodies will further increase the detection target. To solve this problem, many researchers are developing nanobodies. Nanobodies are small antibody fragment from camel; compared to traditional IgG antibodies, nanobodies have the features of smaller molecular weight, easy to produce, good stability, etc.