Do you spend your photon budget wisely? This is a question that a recent paper asks, and answers back with a new technique that produces images that will blow your socks off. Once you put your socks back on, check out today’s image.
Cellular imaging is a constantly evolving field made of biologists on a never-ending quest for higher resolution of structures and faster image acquisition of a living cell. There are several challenges to these demands. For example, cells are not pancake-thin. Current techniques use illumination that leads to background noise in an image due to excited out-of-focus light. In addition, these techniques can cause phototoxic effects on cells, and can photobleach the fluorescent tags used to mark structures. Biologists have addressed these problems by using plane-illumination microscopy, which uses a separate excitation lens positioned orthogonally to the detection objective lens, leading to a more confined excitation of the focal plane. Planchon and colleagues recently improved this technique by using thinner sheets of light to illuminate the sample. The images produced using this Bessel beam plane illumination are remarkable, and allow for very fast 3D imaging of living cells. Images above show filopodia on a HeLa cell (left column), and the membrane ruffles on a kidney cell (right group of images). Purple arrowheads point to vacuole formation by macropinocytosis.
BONUS!! For a movie of the filopodia in the image above, click here. For a movie of the membrane ruffles and vacuole formation, click here. For many other knock-your-socks-off movies, click here.
Planchon, T., Gao, L., Milkie, D., Davidson, M., Galbraith, J., Galbraith, C., & Betzig, E. (2011). Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination Nature Methods, 8 (5), 417-423 DOI: 10.1038/nmeth.1586
Adapted by permission from Macmillan Publishers Ltd, copyright 2011