coscinodiscus(?)

Smokedaddy · Post by **Smokedaddy** » Sat Feb 18, 2017 10:19 am

Thanks Steve ... on another note, I wonder if I've reached the point of what this particular dry objective will resolve with the way I'm using it? Or do I need to make some changes in Zerene (or some other stacking program). Maybe my procedure? I like the diatom. This is a PMax image.

rjlittlefield · Post by **rjlittlefield** » Sat Feb 18, 2017 1:15 pm

It's not likely that any change to your stacking procedure will pull out more detail. PMax is relentless about preserving detail or anything that looks like it. In some cases that can result in artifacts that obscure real detail, but in the image shown here I'm not seeing any of those.

--Rik

Charles Krebs · Post by **Charles Krebs** » Sat Feb 18, 2017 1:40 pm

Just looking at this last one I would say it is excellent.

If there is any real improvement to be had (not saying there is, don't know, this is excellent for your 60/0.85) is is likely to be made with the microscope not the stacking software. From our conversations it sounds like you have your light source and condenser set up properly. The correction collar is really your "friend" when trying to squeeze the most out of a high/dry objective. It really is important to use it carefully. I know you are using a Canon 50D from live mode, so there should be no vibration issues.

There might be a couple "bands" where the step size might have been a little too large... hard to tell if that is what I am seeing or if it is just the diatom.

At best. your effective aperture (in camera "terms") is about f35 (could be considerably smaller, don't know how you have your camera set up). That's just the way is is in high magnification microscopy. So you should not expect the "crispness" you would see in something like a conventional photographic landscape image.

Looking really nice!

Smokedaddy · Post by **Smokedaddy** » Sat Feb 18, 2017 3:21 pm

Thanks Charles. Here's another tried playing with the correction collar and cleaning a few things up with PS. Getting better from what I can see but the center is a little grainy.

Beatsy · Post by **Beatsy** » Sat Feb 18, 2017 3:50 pm

Oooosh! Now that is a cracking Actinoptychus splendens. They are tough to stack because of the fine sieve plate behind the larger pores in the frustule but you've done a grand job here. One of the best I've seen. May be bordering on over sharpening, but it works well with this one.

Smokedaddy · Post by **Smokedaddy** » Sat Feb 18, 2017 4:05 pm

Now that's encouraging especially coming from you. Here's a 100% crop.

Smokedaddy · Post by **Smokedaddy** » Sat Feb 18, 2017 4:16 pm

Charles Krebs wrote:Just looking at this last one I would say it is excellent.

There might be a couple "bands" where the step size might have been a little too large... hard to tell if that is what I am seeing or if it is just the diatom.

Thanks Charles, coming from you that indeed is encouraging. Out of curiosity how could I (like me) make any finer steps. I'm currently going in between the fine focus marks on the dial. Guess I could figure out a way to mount my micron dial indicator on the Optiphot.

Smokedaddy · Post by **Smokedaddy** » Sun Feb 19, 2017 12:42 am

I calibrated the diatom photograph above with ImageJ for microns with my calibration slide and came up with the following measurements. The little circle thing (wart) is approximately 3 microns in diameter, the diatom itself is 120 microns in diameter and the area is 11497.

Then I measured the separation (center to center) on a couple of the dots as shown in the image below via the YELLOW line in the image (if you can see it).

Lastly I measured the diameter of one of the dots, see below.

-JW:

Smokedaddy · Post by **Smokedaddy** » Mon Feb 20, 2017 12:25 am

The latest ...

-JW:

Smokedaddy · Post by **Smokedaddy** » Thu Feb 23, 2017 5:18 pm

Another diatom try today ...

Smokedaddy · Post by **Smokedaddy** » Wed Mar 01, 2017 6:19 pm

I've been trying to resolve the torus area on the circumference and can't seem to get it defined. Any suggestions on setting in Zerene or post processing?

-JW:

rjlittlefield · Post by **rjlittlefield** » Wed Mar 01, 2017 8:01 pm

Smokedaddy wrote:Any suggestions on setting in Zerene or post processing?

I have the usual question: what do individual frames look like?

--Rik

Smokedaddy · Post by **Smokedaddy** » Wed Mar 01, 2017 8:40 pm

... a few random pic(s).

rjlittlefield · Post by **rjlittlefield** » Wed Mar 01, 2017 10:08 pm

Smokedaddy wrote:I've been trying to resolve the torus area on the circumference and can't seem to get it defined. Any suggestions on setting in Zerene or post processing?

Thanks for showing the single frames. I had been concerned that the stacked results might be showing artifacts like parallel bands that had been created by accumulating simpler features across multiple frames. But comparing the composite with the single frame shows that single frame has the same sort of banding.

Likewise the center of the beast looks the same in the stacked composite as it does in the single frames.

So, I don't see that there's any way to improve this result by doing the stacking any different.

I'll leave it up to the micro guys to provide assistance about how the individual frames might improved. Perhaps there's some magic to be done with the illumination. But I haven't worked at all with diatoms, so I'm pretty much clueless in that area.

--Rik

Smokedaddy · Post by **Smokedaddy** » Wed Mar 01, 2017 10:14 pm

Thanks Rik, I was just wondering what I may have done wrong if anything. I probably should have mentioned the stack increments were 0.5 µm.