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Looking for help with specimen contrast / lighting
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JohnyM



Joined: 24 Dec 2013
Posts: 360

PostPosted: Thu May 17, 2018 6:20 am    Post subject: Reply with quote

manu de hanoi wrote:
-are the angled parts blurry just in the final stack or are they blurry too in individual layers ?
-DSLR being closer to the objective as pictured should help you getting good depth of field at the expense of magnification/resolution.
-250 pics for a stack sound like a lot, sometimes less is more (but that depends on you answer to my 1st question)

-Blury in individual layers
-DSLR is literally as close as possible, way below required 160mm
-Number of steps is result of 20x/0.4 providing rougly 3um DOF/slice and subject being almost 1mm thick. According to my math, DOF goes to ~4um with 5x magnification and then this lens is working like a bottle bottom.

[quote]

@ChrisR: Thanks for the ideas.
Gobos sounds interesting. I think i need more magic arms...
Crossed polars might do wonders as they do with epi illumination. Worth a try, but setting up cross pol on 4 jansjos might be a nightmare. Let's see what my shaky hands can do about it.
Coloured? You mean rheinberg? Sadly fancy colors are out of question, unless i end up with converting it to grayscale after (if they help with contrast). I have some Lee gel filters, i'll give it a try.
If you mean hoffman, isn't it basically what angled (oblique) light do in this case?
UV does work, i dont know how well, because specimen degrade blindingly fast, and whole point in going optical instead of SEM is to preserve it.
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