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• Imaging Data Club- Thursday (April 24th @ 6pm)

  Date:

  21 April 2008

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  Dear Group Members,

  Dr Jennifer Waters will lead the Systems Biology Imaging Data Club on Thursday (April 24th, 6pm, in WAB436). You are invited to join us.

  Here is the title and abstract for her talk:

  “Evaluating performance in 3-D fluorescence microscopy”

  Murray, Appleton, Swedlow & Waters
  J. Microscopy Vol 228:390-405

  ABSTRACT

  In biological fluorescence microscopy, image contrast is often degraded by a high background arising from out of focus regions of the specimen. This background can be greatly reduced or eliminated by several modes of thick specimen microscopy, including techniques such as 3-D deconvolution and confocal. There has been a great deal of interest and some confusion about which of these methods is “better”, in principle or in practice. The motivation for the experiments reported here is to establish some rough guidelines for choosing the most appropriate method of microscopy for a given biological specimen. The approach is to compare the efficiency of photon collection, the image contrast, and the signal to noise ratio achieved by the different methods at equivalent illumination, using a specimen in which the amount of out of focus background is adjustable over the range encountered with biological samples. We compared spot scanning confocal, spinning disk confocal, and widefield/deconvolution microscopes and find that the ratio of out of focus background to infocus signal can be used to predict which method of microscopy will provide the most useful image. We also find that the precision of measurements of net fluorescence yield is very much lower than expected for all modes of microscopy. Our analysis enabled a clear, quantitative delineation of the appropriate use of different imaging modes relative to the ratio of out-of-focus background to in-focus signal, and defines an upper limit to the useful range of the three most common modes of imaging.

  Pizza will be served.

  Take care,

  Komal