I try to record on video what i am doing, but how can i share on this forum? It only allows jpgs etc not vids (mp4). I am also very concerned, also for my msc. Methodologuy
Beiträge von Steve_mt
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To prepare 10% ammonium hydroxide solution, should 1 + 9 parts: Conc NH4OH solution & water be mixed or about 1 + 3 parts since conc. ammonia solution is 25-30% concentrated (not 100%). ?
Thanks (i believe the second one! ;-))
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Hi everybody,
@Steve, good news!
But you still have a problem with your optics, probably the condensor is not adjusted. The lines of the eyepiece reticle have a blue shade towards the center, do you see that? And the lines of the reticle are still distorted to the corner, maybe you placed it the wrong way round (resulting in a wrong optical layer for the reticle).
If you spend so much money for buying the best tools, you should also spend some more time to learn better how to use them.
Also you should also spend a few Euro for oil with a guaranteed optimum refractory index, this might improve the results further.
The newer Zeiss and Olympus lenses should have a correction factor very close to 1,000.
... and sell the useless 63x dry and buy a 40x oil instead
Wolfgang
I think the problem is the camera method because I put my Canon G5X over the eyepiece and zoom a bit to fill the field. When I look with my eyes, I do not see the pincushion distortion. For the colour aberration, I see them but maybe I am not very touchy here, but in real life situation, they are not really significant or a big deal at that high magnification. Sometimes I turn the almost moncromatic photos to greyscale.
Saying that, I do not know much about use of the condenser. I usually raise it until I have good contast. Too high I lose the contrast and result in getting a soft image, too down then I have too high contrast and blackened image. Usually, I test with my eye until I have rather good contrast and the blur washes away the background artefacts and dirt. I admit my setup is below average too
- but for now that what I can afford. I miss a trinocular. -
Hello Uwe
yes, that's exactly how I do it. Oil drops directly on the stage micrometer. The idea of putting a drop of liquid and a cover slip on top of it has never even occurred to me. The literature available to me says nothing about this.
best regards,
Andreas
Hmmm.... for me that was the problem, so double check or can you confirm for us that u have no magnification error like what I reported ?
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[...]
... and sell the useless 63x dry and buy a 40x oil instead
[...]
PNF 40x/1,3 Oil: I love it!
Like this ?
Yes not very good optics the 630, I use it mostly for measurements (gives a bit more accurate results). I am not very fond of messing with oil, but maybe I should change mentality...
Do you mean like this?
Carl Zeiss Microscopy, LLC - Objective Assistant - Objective C Plan-Apochromat 40x/1.3 Oil DIC M27
OMG!!! so expensive!!!!
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Dear Andreas, unfortunately, I could not photograph the poorly stained spores very well, but they had one septum. an elongated fusiform shape and usually 4 oil drops. I can confirm that the asci were inamyloid at the tips. Another thing, the ascocarps were growing on bark of a dead olive branch and associated (or on) pyrenomycetes over the bark. All these are indicators for C. sulfurina from the small amount of research I did.
I used Lugol's Iodine solution - I don't know... but it works for me to stain the Asci tips and less dangerous. I do have Melzer's, so if crucial I can test the asci with it again.
I am very disappointed with my image qualities of thid fungus - sometimes I get good images and sometimes I don't. This did not stain very well. I photograph stuff from my Digital camera through the eyepiece. (my microscope is not trinocular
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Flickriver: Searching for photos matching 'Bisporella sulfurina'
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Is "Calycina sulphurea" synonym to "Bisporella claroflava"?LG; Pablo.
Hello Pablo,
yepp.
I worte incorrect "sulphurea" but I meant "sulfurina". Bisporella sulfurina = Calycina sulfurina = Calycina claroflava
all the best,
Andreas
I am lost in nomenclature : Bisporella sulfurina = Calycina sulfurina = Calycina claroflava = Calycella sulfurina ?
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Hi Andreas
He may have put the oil directly on the micrometer object without a cover slip.
Greeting
Uwe
THAT WHAT I USED TO DO!!! when I get the 86 units per 100um (the wrong magnification!)
Now I placed a coverslip over the micrometer fixed coverslip and oil over the second coverslip and I got 98:100. This was so sweaty to arrive to a solution of this mysetrious error. So now my calibrations a slightly screwed when I measured under the x100 but at least not by much (12% error).
I cant understand the physics why a coverslip makes this difference ?!?!
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OMG!!!! It worked!!!!
I have 97-98 reticle units for 100 um!
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I can try micrometer+coverslip over+oil >>> objective and see what I get, but I think I am going to have > 0.17mm working distance
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the x100 (no oil) x 1.05um per reticule decision (calculated magnification x 960)
the x100 (with oil) x 1.17um per reticule decision (calculated magnification x 850)
How have you done this measurement ?
1: test without oil : have you used a cover glas and water inbetween the measurement slide and the objective ?
2: test with oil: have you used a cover glas and water inbetween the measuement slide and the objective ?
This objectives are calculated for 0,17mm coverglas usageBR
Uwe
without oil: Micrometer slide on stage, then no water no cover glass slip (direct visualization)
with oil: Micrometer slide on stage, then droplet of oil on micrometer no cover glass (objective lens in oil)
The micrometer already have a fixed coverslip I don't see why I have to put another one over it and water.
Alles anzeigenthe x100 (no oil) x 1.05um per reticule decision (calculated magnification x 960)
the x100 (with oil) x 1.17um per reticule decision (calculated magnification x 850)
How have you done this measurement ?
1: test without oil : have you used a cover glas and water inbetween the measurement slide and the objective ?
2: test with oil: have you used a cover glas and water inbetween the measuement slide and the objective ?
This objectives are calculated for 0,17mm coverglas usageBR
Uwe
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pilzforum.eu/attachment/363503/I have carried out the micro of the yellow asco. From some observations under the stereo, first impressions are that the pale ones are the same one more or less decolorized because some of them have a bit of yellow pigment in them. The atypical border is perhaps because they are dry ?!
The asci are dextrinoid or stain strongly in IKI except where there are the spores , otherwise inamyloid at the tips; 60-80um long and only 5 um wide. The elongated spores overlap each other in the ascus. The spores are interesting and maybe do not corresponding with C. citrina. They are narrowly fusiform, almost linear with four bands due to the presence of four evenly spaced guttulae. They measuring roughly 10-14 µm x 2-3 µm. They do not stain well in Congo red or IKI. I believe there is a central septum.
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Let me write about the procedure, just in case... I mount the stage micrometer which has a very thin and small circular coverslip fixed to the glass slide, transfer a small drop of oil over its surface, mount, and bring to focus the scale. Then I place the reticule eyepiece (I have 3 of them) and align it so the zero marking is over the end of the stage marking. I count how many reticule markings make a known measurement and calibrate accordingly
the x10 = x10um per reticule division (calculated magnification x 100)
the x40 = x 2.51um per reticule division (calculated magnification x 398)
the x63 = x 1.59um per reticule division (calculated magnification x 628)
the x100 (no oil) x 1.05um per reticule division (calculated magnification x 960)
the x100 (with oil) x 1.17um per reticule division (calculated magnification x 850)
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So sorry for the confusion and enigmatic situation we have here, but now I understand that the problem is only mine
Please note that I do not have any issue with the other objectives, only the x100 oil immersion ones.
To start and solve the issue, the first question is whether my AxioLab RE is finite(160mm) or infinite. The non-oil objectives came with the microscope (great optics) and they have the infinity sign so
[Q1] can we safely assume that the Axiolab RE is an infinite microscope??? I think so!
Answers:
Wolfgang says: If your microscope body is infinite, never use lenses without the ∞ sign.
All my objectives have the ∞ sign. The old oil immersion is 160mm and not the one being tested. Maybe this model can't take x100 objectives?? I doubt it!
Peter asks: You have infinite and finite objectives on the same microscope!
And they all work well for a strange reason - but the one being tested has the infinity sign.
- We don't know what your microscope looks like or what attachments you have. [Answer: link below]
- We don't know what kind of eyepieces you have. [Answer: Zeiss E-PL10]
- We don't know what kind of reticle you inserted. There are different! [Answer: Standard reticle eyepiece from eBay, I don't think it is the problem]
- We don't know where you inserted the graticule. [Answer: Stage micrometre]
- We don't know what oil you use and how you use it. [ Answer: Small drop oil on the stage micrometre and mount. No coverslip ]
[Q2] If there is something wrong with the graticule, eyepieces, reticule.... would the symptom show also on the x630 and x400 objectives?
What's left is the oil to blame perhaps! I don't know anything about its origin, but bought 50mls from Lab supplies who give it in a plastic container.
Really lost now what's the problem??? The speculations of infinity eyepieces on finite microscope is not the issue.
The microscope body is like this (objectives different but they are also with infinity sign):
Carl Zeiss Trinocular Microscope Axiolab | eBay
[Q3] Do you think that the oil-immersion objectives (∞) do not match my microscope ???
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Photos were taken with artificial light so some colour inaccuracies are there, but they are like highlighter yellow, bright. I tried to save some microwork, but i think they are different so I postpone for tomorrow. Would be great if C. sulphurea! Thanks andreas!
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On a fallen small branch of an Olive tree, I spotted some tiny bright yellow ascomycetes which I rolled (ie the entire branchlet) in Aluminum foil and collected it home . Three days later I went to examine the branch and I found two ascomycetes now - (1) the yellow Calycella and (2) another population similar in size to it but gray to mouse gray. Are these some new growth of decolourised specimens of Calycella (because perhaps they where in the dark?)
and also, is C. citrina confirmed from these photos or there are closely related species to be considered (hence I need to run a full micro? )
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Thanks for sharing these Ascomycetes - your microi-mages are amazing.
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Hi Steve,
I do my sequencing with Alfalab.es , Pablo Alvarado has a lot of experience
You will get the results in about 3 weeks , which is quite OKThe price is about EUR 22,00 per sample.
If you like , you can share the sequence data with me, I can help with the interpretation
With rare species its sometimes tricky to interprete the results.
If you are interested contact my by Private Message
BR
UweDear Uwe,
I heard nothing from Unite, so giving them the fair chance to reply, and they did not, I am OK to move on with you. This is the first time to send material for DNA sequencing, hence where should I send the sample to you UWE? - I send you PM as requested
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Hi Steve,
I do my sequencing with Alfalab.es , Pablo Alvarado has a lot of experience
You will get the results in about 3 weeks , which is quite OKThe price is about EUR 22,00 per sample.
If you like , you can share the sequence data with me, I can help with the interpretation
With rare species its sometimes tricky to interprete the results.
If you are interested contact my by Private Message
BR
UweDear Uwe,
I herad nothing from Unite, so giving them the fair chance to reply, and they did not, I am OK to move on with you. This is the first time to send material for DNA sequencing, hence where should I send the sample to you UWE? - I send you PM as requested
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It is a petiole (stalk) of a leaf of Rubus ulmifolius, but I have also collected the species from legume of decaying carobs
regards Steve!
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The Axioscope 20 should be infinite, is it?
Yes, it is!
Sorry folks - My microcope is AxioLab RE not Axioscope 20
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The Axioscope 20 should be infinite, is it?
Yes, it is!
Sorry folks - My microcope is AxioLab RE not Axioscope 20
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The other old x100 oil eyepiece behaves the same. In oil mount 100um gives 85 reticule units and was shown in the photo.
This is a Zeiss NeoFluar 100\ 1.30 Oel 160\- code 48600960
Please not that my Microscope Is AxioLAB RE
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This is the one I am testing right now:
ZEISS Plan-NeoFluar
100x / 1,30 Oil
1018-595 (1066-987)
on the other side
ZEISS Plan-NeoFluar
100x / 1,30 Oil
∞ 0,17
I took photos by digital camera from the eyepiece.
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Daturamyces
