Crosstalk (aka bleed-through or crossover) is a serious problem when analysing multichannel microscopy images, because you record some signal as a certain dye when it really comes from a different one. With Huygens CrossTalk Corrector you can quickly estimate and correct crosstalk between multiple fluorescent channels simultaneaously. We highly recommend to correct for crosstalk before performing deconvolution, and to consider also chromatic aberration correction. Especially, if you continue with image analysis that involves multiple channels, like for example colocalization analysis.
Mitochondrial (TMRM) red signal is also detected as signal in the green Lipid Droplet (BODIPY) channel due to crosstalk (left side of the image). This crosstalk gives a wrong representation of the lipid droplet structures and will have a serious impact on subsequent analysis. The right side of this image has been corrected for crosstalk with Huygens Crosstalk Corrector and shows both channels or only the green lipid droplets. This spinning disk image was kindly provided by Kevin Knoops (Microscopy CORE Lab), Sabine Daemen and Matthijs Hesselink, Maastricht University, The Netherlands.
Automatic and Easy
Quickly access if your image suffers from crosstalk using automated estimation, and correct the crosstalk with one additional click of a button.
Use the coefficient matrix to see and change the crosstalk coefficients of all channels (up to 32).
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See more: Images in the field of NeuroBiology
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