A quick guide to light microscopy in cell biology

A few months ago, I was asked to write a introduction for newcomers to light microsopy for Molecular Biology of the Cell (MBOC). That paper is now out, and I’m pretty pleased with how it came out. It’s designed to provide a brief introduction to a graduate student or postdoc in cell biology who’s new to microscopy and wants a brief orientation to the field.

I’ve taken to turning down offers to write reviews because in general, I don’t think the impact is worth the effort required to write them (I also don’t like signing my copyright over to the publisher). I agreed to write this one because I think MBOC is a good journal – they focus on good science and all papers are freely accessible two months after publication and the Technical Perspective fills a useful role of providing orientation to newcomers in a field. I actually had a lot of fun writing the paper – it was nice to be able to sit down and write without having constantly refer to the literature or to data.

4 thoughts on “A quick guide to light microscopy in cell biology

  1. Hi Kurt,

    I would like to read it. The article is not available as open source. Can I get a copy please.

    • Hi Stan –
      I sent a copy to you separately; the paper will also be freely available in two months.

  2. I love your website. I enjoyed the guide to light microscopy as well. I have a question about table 1. The table suggests that widefield microscopy is slower and less sensitive than spinning disk microscopy. Why? I thought that the major advantage of widefield microscopy is speed and sensitivity.

    • That table was put together based on my experience with our scopes, and so applies to scopes as you see them usually configured. You certainly could put together a widefield system as fast as a spinning disk system, for example. Sensitivity is a bit of a trickier issue. In principle, widefield plus deconvolution is more sensitive than spinning disk (see http://www.ncbi.nlm.nih.gov/pubmed/18045334). However, you seldom see widefield scopes equipped with EMCCDs, whereas spinning disks often are, and spinning disk seems to be more sensitive than widefield without deconvolution (at least that is what we observe, though we have not carefully measured it). The proper comparison is fairly complicated, because in order to get good deconvolution you need to take a fair number of out of focus planes, which slows down your acquisition rate, so we have had users who have chosen spinning disk over deconvolved widefield because the combination of speed and sensitivity of spinning disk confocal was tough to beat.

      I probably should have made that table look less definitive, because configuration matters a great deal. Unfortunately, in such a short introduction there wasn’t time to go into all these details.

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