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Less damaging laser powers and higher detection dynamics yield more valid results in live cell imaging.
High dynamic range (HDR) imaging is well known from still photography. In a live imaging system, it seems to be a bad idea to obtain multiple images and expose your sample to repeated laser illumination. However, in biological samples the fluorescence dynamics are often bigger than the capacity of the detection system.
 Non-neuronal retina structures labelled with Cyano-dyes, very high signal dynamics; left conventional, right HDR acquisition. Sample: F. Tatin, Cancer Research, London, UK. |
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| Non-neuronal retina structures labelled with Cyano-dyes, very high signal dynamics; left conventional, right HDR acquisition. Sample: F. Tatin, Cancer Research, London, UK. |
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The ROI-HDR mode of the LSM 780 and LSM 710 allows you to image the weak and bright portions of a frame in the first shot (or line) and then acquire the weak signal subregions (ROIs) with a second shot to amplify them. The software lets you choose between an adapted mode or a linear mode which even allows quantitative analysis of the signals.
The result is an increased dynamics in detection, without superfluous laser exposure to the sample. Without saturation, for example, a bright soma of a nerve cell can be imaged together with the weak and faint dendrites so it’s altogether easier on your specimen.
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| The number of frames and illumination variations can be set freely for the HDR acquisition. |
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