My first diatom stack

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Wes
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My first diatom stack

#1 Post by Wes » Fri May 31, 2019 10:22 am

Here I manually took a few images of an unknown diatom in different z-planes and stacked them together in Photoshop. I know there is a separate ID forum but if you have a clue what this diatom is please share :)
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63/1,4 Planapo, DIC
63/1,4 Planapo, DIC
diatom_stack.jpg (248.03 KiB) Viewed 5598 times
Zeiss Photomicroscope III BF/DF/Pol/Ph/DIC/FL/Jamin-Lebedeff
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75RR
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Re: My first diatom stack

#2 Post by 75RR » Fri May 31, 2019 11:06 am

Nice to see another brave soul using the Zeiss 63x/1.4

That looks like some serious magnification.

Did you remember to measure it? Size is an important variable in ID

Location and environment also help. i.e. freshwater, brackish, marine ...

Nice work!
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

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Wes
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Re: My first diatom stack

#3 Post by Wes » Sat Jun 01, 2019 7:02 am

Nice to see another brave soul using the Zeiss 63x/1.4
Is there some kind of general reluctance to use this objective?

Anyway the diatom came from a freshwater pond in the nearby park. Unfortunately I did not measure it. I have a stage micrometer but not an eyepiece graticle. It should still be possible to do without the graticle by taking an image of the stage micrometer and calculating the size from pixel distance.
Zeiss Photomicroscope III BF/DF/Pol/Ph/DIC/FL/Jamin-Lebedeff
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MicroBob
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Re: My first diatom stack

#4 Post by MicroBob » Sat Jun 01, 2019 7:22 am

Wes wrote:but not an eyepiece graticle.
Hi Wes, you can print one with a laser printer on overhead projection film. Under magnification these laser prints don't look too nice but as a scale this works acceptably. Ink jet might even be better.

So there is no reason to deny us the measurement! :mrgreen:

Bob

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75RR
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Re: My first diatom stack

#5 Post by 75RR » Sat Jun 01, 2019 7:59 am

Is there some kind of general reluctance to use this objective?
It can be a little finicky, even seems to have a mind of its own on occasion.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

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Wes
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Re: My first diatom stack

#6 Post by Wes » Sat Jun 01, 2019 10:13 am

MicroBob wrote:
Wes wrote:but not an eyepiece graticle.
Hi Wes, you can print one with a laser printer on overhead projection film. Under magnification these laser prints don't look too nice but as a scale this works acceptably. Ink jet might even be better.

So there is no reason to deny us the measurement! :mrgreen:

Bob
Well I did that yesterday but for a different reason. I had to center one element in the headrig of my Phomi so I printed evenly spaced black dots on an overhead projection film. Well my approach failed so I used a normal histology slide to get the centering but I can use your tip and produce a reticle. You wouldn't happen to have a template I could borrow, would you? :D
75RR wrote:
Is there some kind of general reluctance to use this objective?
It can be a little finicky, even seems to have a mind of its own on occasion.
Very interesting that you mention this. The one used to get this image is delaminated. I have another one that looks overall superior i.e. no delamination or other visible issues. To my surprise the delaminated planapo gives much better contrast and color correction than the seemingly better one. If I focus on an object and have in the middle of the field of view it gets shifted quite a bit when I switch to the other planapo so the 'better' one is probably off-centered. No idea why or what I can do about it other than return it. The other thing I haven't been able to figure out is why the DIC slider works much better when I flip it i.e. the label pointing towards the base of the microscope. Not only is contrast better but the dark to light gradient is almost non-existent. In its normal position the DIC slider gives a very sharp dark-to-light gradient and overall poor contrast.
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Hobbyst46
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Re: My first diatom stack

#7 Post by Hobbyst46 » Sat Jun 01, 2019 10:28 am

I find the 63X1.4 Planapo phase contrast easier to use than the 100X1.3, and it yields excellent images, so perhaps just the DIC version is finicky.

MicroBob
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Re: My first diatom stack

#8 Post by MicroBob » Sat Jun 01, 2019 12:02 pm

Here is a PDF with eyepiece reticules: https://drive.google.com/open?id=1bVYdt ... o6722zLXRQ

It works with 80% of the eyepieces I tested. In most cases there is a convenient spot to put the reticule on. Real measuring eyepieces provide focusing to get a perfectly sharp image of the reticule. In my experience this is not needed when you use them with glasses if necessary.

Bob

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75RR
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Re: My first diatom stack

#9 Post by 75RR » Sat Jun 01, 2019 12:55 pm

I was using a no name 10x eyepiece with reticule to measure animalcules that worked well, only problem was that my eyepieces are 12.5x so I had to shut one eye when using it.

Ended up getting a 12.5x eyepiece, which unfortunately had a grid reticule rather than the one I needed so I bought this: https://www.ebay.co.uk/itm/323274970066 to replace it.

Had to sand it down from 17.8 (the smallest they had) to 17mm

Lines and numbers are clear and sharp. Very happy with it.

Photo: Eyepiece held up to phone camera with computer screen as backdrop
Attachments
reticule.jpg
reticule.jpg (150.08 KiB) Viewed 5483 times
12.5x eyepiece.jpg
12.5x eyepiece.jpg (62.42 KiB) Viewed 5504 times
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

Hobbyst46
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Re: My first diatom stack

#10 Post by Hobbyst46 » Sat Jun 01, 2019 3:38 pm

75RR wrote:... had a grid reticule rather than the one I needed so I bought this: https://www.ebay.co.uk/itm/323274970066 to replace it.
Had to sand it down from 17.8 (the smallest they had) to 17mm
Lines and numbers are clear and sharp. Very happy with it.
Thanks 75RR for the link. I followed suit.

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Re: My first diatom stack

#11 Post by MicroBob » Sat Jun 01, 2019 5:40 pm

75RR wrote:Very happy with it.
There it is: A satisfying chinese product! :D If one could depend more on the quality it would become difficult for us to compete...
I have a chinese object micrometer: Also very satisfying for my needs (haven't checked the absolute accuracy)

Bob

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Re: My first diatom stack

#12 Post by Hobbyst46 » Sat Jun 01, 2019 6:26 pm

MicroBob wrote:
75RR wrote:Very happy with it.
I have a chinese object micrometer: Also very satisfying for my needs (haven't checked the absolute accuracy)Bob
Hi Bob, I have compared an inexpensive Chinese stage micrometer against a Zeiss metal-based stage micrometer (bright lines on opaque background), and the deviation - if any - is certainly less than 1%.

MicroBob
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Re: My first diatom stack

#13 Post by MicroBob » Sat Jun 01, 2019 8:53 pm

Hi Doron,
this is what I would expect and I wouldn't worry about 1%. Fit for purpose, acceptable price and available for everybody. It is great to have a common test plate that arrives at you doorstep without hunting ebay auctions etc.
To save costs the manufacturers tend to just barely deliver what the customer needs. So I would expect that even a Zeiss object micrometer will have a certain tolerance. The process itself makes it difficult to obtain crazy precision.
As far as I know the production process is quite dangerous, nothing for the kitchen table. I have a grid plate made by Peter Höbel as a kind of comparable standard for objective evaluation, made just before the cheap chinese object micrometers became available. He has the equipment to safely go though this process but didn't like it much. :twisted:

The historical development of the scientific measuring methods, machine tools and measuring tools was tightly entwined. How to make a precision spindle without a lathe with a precise lead screw? Pieter Harting, a dutchman, wrote in his book "Das Mikroskop" (german translation) about stage micrometers. https://ia800206.us.archive.org/34/item ... 073563.pdf
To make the spindles they took wire (first precision material mass made) and wound it around a core of known diameter. This gave in theory a nice uniform lead screw of known pitch. Problem was not all manufacturers calculated the elongation of the wire, caused by the force to pull it around the core. 8-)

Bob

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actinophrys
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Re: My first diatom stack

#14 Post by actinophrys » Sun Jun 02, 2019 4:35 am

Very nice, Wes. For the record, this looks like some type of Amphora, which characteristically have lunate valves tilted to the same side of the cell; the empty band in the middle is the girdle.

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Wes
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Re: My first diatom stack

#15 Post by Wes » Sun Jun 02, 2019 11:31 am

MicroBob wrote:Here is a PDF with eyepiece reticules: https://drive.google.com/open?id=1bVYdt ... o6722zLXRQ

It works with 80% of the eyepieces I tested. In most cases there is a convenient spot to put the reticule on. Real measuring eyepieces provide focusing to get a perfectly sharp image of the reticule. In my experience this is not needed when you use them with glasses if necessary.

Bob
Thanks Bob, I will give it a try.
75RR wrote:I was using a no name 10x eyepiece with reticule to measure animalcules that worked well, only problem was that my eyepieces are 12.5x so I had to shut one eye when using it.

Ended up getting a 12.5x eyepiece, which unfortunately had a grid reticule rather than the one I needed so I bought this: https://www.ebay.co.uk/itm/323274970066 to replace it.
Thinking about the eyepiece you posted made me realize I need to upgrade my Cpl eyepieces if I want to get the most out of those planapochromats I've invested in. For the camera adapter I am using the good old 10x Kpl which gives good results.
actinophrys wrote:Very nice, Wes. For the record, this looks like some type of Amphora, which characteristically have lunate valves tilted to the same side of the cell; the empty band in the middle is the girdle.
Awesome thanks! It does look a lot like Amphora I agree :D
Zeiss Photomicroscope III BF/DF/Pol/Ph/DIC/FL/Jamin-Lebedeff
Youtube channel

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