Rik - great idea to use Nth frame - had not been aware of that feature. I assume, however, that the fraction of DoF for step size used will need to be be different with different subjects and level of detail (not to mention lighting). But that just makes the task more of a challenge and the rewards ...

On a related note, I have a MIty 100 for sale and someone asked me for the suggested step size. This objective has an NA of .70 and the formula Zerene/Rik suggests is .00055/(NA*NA), which calculates out to .00055(.70*.70), or .00055/.49, which is .00112, so I guess 1 um would be about right. The fe...

Thanks very much for the reply and cleaning tips.

Thanks, Rik. Interesting assessment. Is there a known cutoff for magnification factor above which you think fractional DoF per step becomes essential?

"I bathe insects in alcohol, then dry them with air spray." I'm trying to develop some best practices for cleaning my insect specimens. Couple of questions: Do you use 70% or 90% alcohol? Do you rinse them in water before air drying? What type of device do you use for the air drying? And what do you...

Thanks, Macrero. I am always trying to improve my technique - a never ending challenge ;-) And this was the first I had heard of using anything smaller than the DoF. I live by Rik's table (2-C) in the Zerene manual, which says : "Determine DOF (step size) [sic] from Numerical Aperture (NA)," so I ha...

"Did the "poster" post some proof"
There was noting polemical about the comment. It was just a suggestion. I'm trying it currently to determine if it will help my high magnification stacks.

Thanks very much. (Probably the reason I am having trouble with 20x and 50x.) Never too late to learn!

This is probably very old news to most, but I just saw a post on FB recommending that the step size for any given stack should be at least the DoF/3. Is that the "industry" standard? (Poster also recommends DoF/6 if printing large prints.)

Well as Lou noted, sometimes the first and often the last (I usually throw in a few extra on front and both due to backlash concerns) are often OOF. So that may be part of the issues I have been running into. I will have too rethink how I choose the frames to export to ZS... Thanks for the clarifica...

"Alignment in ZS is done by choosing one image as master"

Curious how the master image is selected - I occasionally start my stack list with a frame that may not have an area entirely in focus - sometimes it's hard to tell... Is the master frame the first frame in the list, by any chance?

I'd like to contest the claim of "for optimum performance, always use .17mm". Always is a very strong word. Thanks, M_C. I think the 13- 17mm variety will work best for me. I am only looking at dried chemical solutions - very thin - and prefer large ..x 60mm so I can get the most coverage over the ...

Thanks very much, M_C. I do use 40x (.65) occasionally, but don't often know in advance what magnification I will want for any given subject. And since I am heating the solution, it would be tricky to use a coverslip from the start, rather than put the solution onto the slide and then cover.... I wi...

viewtopic.php?p=198514#198514 and in the surrounding thread.

From the link above: "For optimum performance, always use .17mm .."
I tried to find .17 online but I only see ".13mm - .17mm" and ".17 - .25," etc. So which might be best?

Thanks, Rik.

Thanks much for the detailed reply. I am always looking for highest resolution when photographing the crystals, and my objectives vary from 4x to 40x (will check the NA - .1 to .65, I believe). So maybe best to try different thicknesses and see what works with what. The coverslip *is* the front elem...