Nitzschia sigma

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75RR
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Nitzschia sigma

#1 Post by 75RR » Sat Nov 23, 2019 10:38 am

Planapo 63x/1.4, DIC, 155µm length - 8µm width, stacked and stitched in Photoshop

This is from a Klaus Kemp 8 Form slide

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MicroBob
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Re: Nitzschia sigma

#2 Post by MicroBob » Sat Nov 23, 2019 5:00 pm

Hi Glen,
nice image! This is probably one of the diatoms that end up in broken pieces in many cases after cleaning.

Bob

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Wes
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Re: Nitzschia sigma

#3 Post by Wes » Sat Nov 23, 2019 5:30 pm

Thats really good! Excellent contrast and resolution, thanks for sharing.
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Re: Nitzschia sigma

#4 Post by 75RR » Sat Nov 23, 2019 5:37 pm

Thanks MicroBob,

I am afaid I can't take any credit for its condition as I had nothing to do with its cleaning or mounting.

It is from a Klaus Kemp 8 Form Test slide

Here is some info on the diatom: https://diatoms.org/species/nitzschia_sigma1
.
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Kemp-II.jpg
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Last edited by 75RR on Sat Nov 23, 2019 5:51 pm, edited 1 time in total.
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Re: Nitzschia sigma

#5 Post by 75RR » Sat Nov 23, 2019 5:39 pm

Thanks Wes
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Re: Nitzschia sigma

#6 Post by Wes » Sat Nov 23, 2019 5:52 pm

Out of curiosity do you see the striae in Amphipleura pellucida with the 63x planapo? It might be the right subject to also test the effect of condenser oiling on resolution.
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Re: Nitzschia sigma

#7 Post by 75RR » Sat Nov 23, 2019 6:05 pm

Wes wrote:
Sat Nov 23, 2019 5:52 pm
Out of curiosity do you see the striae in Amphipleura pellucida with the 63x planapo? It might be the right subject to also test the effect of condenser oiling on resolution.
Funny you should ask, had a look today while I was imaging the Nitzschia sigma but had no luck.

I seem to recall being able to resolve the puncta previously but did not manage to do so today.

Which leads me to suspect that it requires a more subtle setup.
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Re: Nitzschia sigma

#8 Post by PeteM » Sat Nov 23, 2019 6:08 pm

Nicely done. Thanks for sharing it.

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Re: Nitzschia sigma

#9 Post by ImperatorRex » Sat Nov 23, 2019 9:05 pm

Great Photos 75RR!

Maybe you need to try one of the older Zeiss Objectives with n.A. of incredible 1.6:
PlanApo 100/1,6 Imm or the 74/1,6 ;)

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Re: Nitzschia sigma

#10 Post by 75RR » Sat Nov 23, 2019 9:44 pm

-
Thanks PeteM
Maybe you need to try one of the older Zeiss Objectives with n.A. of incredible 1.6:
PlanApo 100/1,6 Imm or the 74/1,6
Love to try, though I fear I would not be able to do them justice
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Re: Nitzschia sigma

#11 Post by 75RR » Sat Nov 23, 2019 9:48 pm

-
Thanks PeteM
Maybe you need to try one of the older Zeiss Objectives with n.A. of incredible 1.6:
PlanApo 100/1,6 Imm or the 74/1,6
Love to try, though I fear I would not be able to do them justice
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
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Re: Nitzschia sigma

#12 Post by Wes » Sat Nov 23, 2019 10:03 pm

ImperatorRex wrote:
Sat Nov 23, 2019 9:05 pm
Great Photos 75RR!

Maybe you need to try one of the older Zeiss Objectives with n.A. of incredible 1.6:
PlanApo 100/1,6 Imm or the 74/1,6 ;)
NA of 1,6?? I had never heard of that till now. I know some of the modern 100x planapos used for TIRF microscopy deliver NA of 1,46. Can you provide a bit more info, this sounds remarkable.
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Re: Nitzschia sigma

#13 Post by 75RR » Sun Nov 24, 2019 6:50 am

Wes wrote:
Sat Nov 23, 2019 10:03 pm

NA of 1,6?? I had never heard of that till now. I know some of the modern 100x planapos used for TIRF microscopy deliver NA of 1,46. Can you provide a bit more info, this sounds remarkable.
I assumed that those were the objectives used in the famous 1888 catalogue ... turns out they used a 1.3NA

This is the blurb under the image as translated by google:

Pleurosigma angulatum. Enlargement: 4900.
Objective: Apochromat 2.0 mm, 1.30 aperture; Projectios Ocular 4.
Illumination: central, solar image projected into the object plane by Apochromat 16.0 mm, 0.30 Apert.


Here is the link: https://ia800202.us.archive.org/9/items ... 109582.pdf

Which leaves me with the same question as yours
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Pleurosigma angulatum. Magnification- 4900.v.jpg (168.28 KiB) Viewed 5808 times
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Re: Nitzschia sigma

#14 Post by 75RR » Sun Nov 24, 2019 7:09 am

Found the link!
"All these photos were taken with the Zeiss lens 1/10 inch, nA 1.63. Monochromatic sunlight. Special compensation eyepiece 12. Condenser 1.6. Inclusion in a medium with refractive index 2.4. Coverslips and flint glass slides with a refractive index of 1.72."
Image is in the post above the one I linked to

viewtopic.php?f=15&t=6806&p#p60731
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Re: Nitzschia sigma

#15 Post by Hobbyst46 » Sun Nov 24, 2019 8:39 am

Thanks 75RR for the post of the 4900x enlargement. It is impressive, and as previously discussed, was achieved by means of illumination, condenser, media and slide optimization; likely a 1.6NA objective alone would not provide such nice details.

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Re: Nitzschia sigma

#16 Post by MichaelG. » Sun Nov 24, 2019 9:04 am

Hobbyst46 wrote:
Sun Nov 24, 2019 8:39 am
Thanks 75RR for the post of the 4900x enlargement. It is impressive, and as previously discussed, was achieved by means of illumination, condenser, media and slide optimization; likely a 1.6NA objective alone would not provide such nice details.
I agree ... and I think our inability to match such results is, in part, attributable to the scale of our image sensors.
That image was [if I understand correctly] produced on the equipment shown in plates XII and XIII.

There was recently a rather fruitless discussion here, about pixel size and ‘resolution’
See message #15 et seq here: viewtopic.php?f=9&t=5907#p72498

That Zeiss equipment is using a very large “sensor” ... so the grain in the emulsion is providing the equivalent of very small pixels. Q.E.D.

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Re: Nitzschia sigma

#17 Post by MichaelG. » Sun Nov 24, 2019 12:03 pm

Regarding the 4900x image : It is interesting to speculate how good the original silver image must have been.
It is very clear that the image we are seeing has been considerably degraded by the scanning process ... and the published image was, of course, already poorer than the original.

MichaelG.
detail from the pdf
detail from the pdf
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Re: Nitzschia sigma

#18 Post by Wes » Sun Nov 24, 2019 8:03 pm

75RR wrote:
Sun Nov 24, 2019 7:09 am
Found the link!
"All these photos were taken with the Zeiss lens 1/10 inch, nA 1.63. Monochromatic sunlight. Special compensation eyepiece 12. Condenser 1.6. Inclusion in a medium with refractive index 2.4. Coverslips and flint glass slides with a refractive index of 1.72."
Image is in the post above the one I linked to

viewtopic.php?f=15&t=6806&p#p60731
Thanks!

Its impressive to say the least, especially considering the time when it was realized.
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Re: Nitzschia sigma

#19 Post by MicroBob » Sun Nov 24, 2019 10:15 pm

Hi together,
as far as I know these Zeiss n.a. 1,6 objectives were used with a different immersion medium and had very fragile front lens mounting. The n.a. 2,4 mountant must have been arsenic yellow medium. These objectives were probably available until the TEM and SEM became available. Today it would probably be possible to go beyond n.a. 1,6 in light microscope objective and condensers but nobody cares. What is used in confocal microscopy?

Bob

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Re: Nitzschia sigma

#20 Post by Hobbyst46 » Sun Nov 24, 2019 10:42 pm

MicroBob wrote:
Sun Nov 24, 2019 10:15 pm
Hi together,
as far as I know these Zeiss n.a. 1,6 objectives were used with a different immersion medium and had very fragile front lens mounting. The n.a. 2,4 mountant must have been arsenic yellow medium. These objectives were probably available until the TEM and SEM became available. Today it would probably be possible to go beyond n.a. 1,6 in light microscope objective and condensers but nobody cares. What is used in confocal microscopy
Bob, in "simple" confocal microscopy the resolution is still described in terms of the Abbe or Raleigh rules. And the same objectives are used (say, NA up to about 1.4, mostly). The point is, that a high NA is favored in confocal because it gathers more light; and confocal is essentially fluorescence microscopy. The basic advantage of confocal is removal of out of focus images.
There are more sophisticated, super-resolution confocal microscopes, where resolution is not limited by diffraction; however, as much as I remember, super-resolution is achieved not by particularly high NAs but by other means..

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