Microscopy Madness in Bangalore

Published in Protocols & Methods

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Editorial note: Welcome (and a big thank you) to our first ever guest bloggers – Professor Hari Shroff and Dr Abhishek Kumar! Their post perfectly illustrates why we at Nature Protocols are so passionate about helping to ensure detailed technical knowledge and experience is transferred from one lab bench to another, from senior to junior researchers, across the globe. Enjoy their story – and be inspired by the achievements of the students!

A guest blog post by Hari Shroff and Abhishek Kumar

Optical imaging is experiencing an explosion, and not a month goes by without the publication of some new microscopy method. This can make it difficult to keep up, even for a lab like ours1, whose business is to create new imaging tools for use in biology. Even more challenging is the lag between the development or prototype of a new technology, and deployment of the tool in a form that biologist can easily use. Technology translation can take years and commercial adoption of a new method even longer- if it happens at all.

We are thus grateful for the opportunity to publish a do-it-yourself, easier-to-use implementation of our dual-view selective plane illumination microscope (diSPIM)2, a high-resolution light-sheet system, in Nature Protocols3. Apart from employing fiber-coupled excitation (making it easier to use a variety of commercial laser sources, and improving the flexibility and alignment of our device) and describing the use of GPU-ready processing software (speeding up image processing tremendously compared to our earlier efforts2), the protocol provides step-by-step instructions on construction, alignment, and imaging at a level of detail impossible to provide in our previous publication.

Of course, there’s no better test of a method than trying to put it into practice – which we were lucky to do recently at the 6th annual microscopy course in Bangalore, India at the National Center for Biological Sciences (NCBS). The course runs for one week, and students alternate between lectures and labs. Working days are long, with most days 12-14 hours. At the end of the course, students pick up a short independent project, putting what they learned at the course into practice.

When we arrived in Bangalore, NCBS staff had pieced together much of a diSPIM, based on an earlier version of the Protocol that we had sent them. We spent most of the week checking various components and fixing small issues. By the time the final weekend rolled around, we decided to rip it up and build it again from scratch – a great student project. In about 6 hours, course participants were able to assemble and align a functional microscope (well, almost functional as we had some trouble with the piezos…) by following our Protocol – perhaps the first light-sheet system in India but certainly the first diSPIM.

The results of their labors (a total time of ~6 hours) are entertaining (sped up ~65x and condensed to ~4 minutes), and can be viewed here.  We hope you enjoy it – for maximizing the audio-visual experience, we recommend turning your speakers up!

  1. http://www.nibib.nih.gov/about-nibib/staff/hari-shroff
  2. Wu, Y. et al. Spatially isotropic four-dimensional imaging with dual-view plane illumination microscopy. Nat Biotechnol. 31, 1032-1038 (2013). DOI: 10.1038/nbt.2713
  3. Kumar, A. et al. Dual-view plane illumination microscopy for rapid and spatially isotropic imaging. Nature Protocols 9(11), 2555 – 2573 (2014). DOI: 10.1038/nprot.2014.172

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