| Rosemary, Thank you for the rapid reply! Often when GFP is weak (ex488/em522), such as in mitochondrial targetting, the chloroplasts often in overlay become yellow as the autofluorescence becomes very noticeable. Furthermore, if there is GFP in the chloroplasts which is not quite bright, it can be very difficult to demonstrate its presence to reviewers. I always set up against wild type material, and I'm always looking for ways to explain to users what is going on with their "green" chloroplasts which should be red! Thanks. Christian |
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