GFP tagging virus was injected in the rat brain and after 3 weeks, rat was purfused.
I sliced brain tissue in 1mm thickness and
cleared with Rapiclear1.47 for several days.
Also, mounted the sample with iSpacer and Rapiclear solution.
However, when I tried to image the sampel with confocal microscopy (olympus) 100x in depth,
focus was not fixed and the result was blurry-greenish image.
I am not sure whether the sample was a problem(not cleared enough) or virus was the problem.
White matter was cleared in a yellowish color but the gray matter was very evident.
Could you share your opinion on this?