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PostPosted: Fri Jul 19, 2019 9:04 pm 
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Hi all, I just had a quick try at stacking a 4-image set of images from the wonderful 60x collar-corrected objective, the result is very pleasing indeed. This is from a permanently-mounted & stained slide of sectioned (not whole) Sonchus.sp pollen-grains, which I find are a real struggle to stack well....

Anyway, the method I used, not sure why, just 'seeing what happens' I think, was to ste the collar to 0.17µ and focus the first layer of the stack. Then, instead of subsequently focusing the further 3 layers with the BX40's fine-focus in the usual way, I used the collar to 'drill-down' for the 3 further layers of focus. I doubt this method makes any difference but thought I practice with the 60x a little. I have no similar stack mad by using the 'scope's fine-focus for each layer - still, maybe I'll do that tomorrow!

A few images, including the 4 layers of the stac and of course the stack itself, made using Helicon Focus and the 'Depth-Map method of stacking with a width setting of 30 - always goos settings I find for my plaant slide stacks.
I also made what I call a 'pseudo SEM' image which I sometimes indulge-in as I really like the 3-D look, see what you think. It's made simply with PSE v9 by first inverting the colours (not the orientation!) of the 'normal' stacked image, then converting that to monochrome and that's all really - I love the effect, especially with pollen exines such as here.

The first 4 images are the constituent layers of the stack, each layer focused with the objective's collar,
Stack layer 1 of 4
Image
Layer 2 of 4
Image
Layer 3 of 4
Image
Layer 4 of 4
Image

This is the stack, edited lightly in PSE v9,
Image

This is my rather indulgent 'simulated SEM' edit.....
Image

A very enjoyable quick outing with the 60x - which I've also given a light clean as it was rather grimy around the collar.

Hope you like them! :D

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John B


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PostPosted: Fri Jul 19, 2019 9:19 pm 
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Hello John B.,

These are great photos.. I am glad your new microscope works well for you..

BillT


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PostPosted: Fri Jul 19, 2019 9:31 pm 
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One small step for Man ...
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MichaelG.

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PostPosted: Sat Jul 20, 2019 3:13 am 
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mrsonchus wrote:
Then, instead of subsequently focusing the further 3 layers with the BX40's fine-focus in the usual way, I used the collar to 'drill-down' for the 3 further layers of focus.


Hi John,
the 100% method:D would be to focus with the fine adjustment of the scope, then choose the best collar setting. Before you settle on the collar focussing method I would sugest to compare the result to fine focus adjustment and Fine focus adjustment + collar afjustment.
I would guess that the collar is handy but not ideal to focus through the stack. On my old Zeiss objectives the effect of the collar adjustment isn't obvious and it takes careful observation to find the best setting. So pictures might be the better proof.

Bob


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PostPosted: Sat Jul 20, 2019 6:44 am 
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MicroBob wrote:
mrsonchus wrote:
Then, instead of subsequently focusing the further 3 layers with the BX40's fine-focus in the usual way, I used the collar to 'drill-down' for the 3 further layers of focus.


Hi John,
the 100% method:D would be to focus with the fine adjustment of the scope, then choose the best collar setting. Before you settle on the collar focussing method I would sugest to compare the result to fine focus adjustment and Fine focus adjustment + collar afjustment.
I would guess that the collar is handy but not ideal to focus through the stack. On my old Zeiss objectives the effect of the collar adjustment isn't obvious and it takes careful observation to find the best setting. So pictures might be the better proof.

Bob


Hi Bob, yes the 'collar stack' was just a tinker. Just thought I'd see what happened....
As I understand it as the Nikon web pages say, the collar is best set to 0.17 and the 'scope focused, then adjustments with the collar to see if improvement may be made. I think the best method for stacking with a collar-corrected objective would be to initially find the best collar position for layer 1 and leave it there, then to use the 'scope for subsequent layers in the usual way.
Adjusting the collar and the fine-focus for each layer is I'm sure too disruptive (potentially certainly) of the strict layer sequence in one direction only, required for stacking. Fine tuning the collar involves both directions (i.e. up and down) of focal adjustment of both collar and fine-focus according to the Nikon site I linked to above.

My method with this fine objective will be to fine-tune the collar on the 1st layer of any stack, then focus following layers with the fine-focus in the usual way - it was interesting to use the collar to stack but just a 'play-around' with the collar really....

The translucency of mounted and stained transmition-illuminated slides in BF is always a real challenge to stacking, I don't stack often, and very rarely beyond about 6 layers with these slides.

The precision and detail of the BX40 is a real treat for me - each slide is almost like a new slide - new details keep popping-out as I re-visit some of my old favourites!

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PostPosted: Sat Jul 20, 2019 12:11 pm 
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billbillt wrote:
Hello John B.,

These are great photos.. I am glad your new microscope works well for you..

BillT


Hi Bill, thanks my friend, good to hear from you. :)

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