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Hi from northern Germany

 
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pbraub



Joined: 02 Feb 2018
Posts: 37

PostPosted: Thu Dec 13, 2018 4:10 am    Post subject: Hi from northern Germany Reply with quote

Dear photomacrography and amateurmicrography community,

after a sometime reading and a shorter time posting I feel that it is my time to introduce myself.

I am working in pathology and by this am able to enjoy microscopy most of my working day - I hope this does not disqualify me from the amateurmicrography forum. Besides my day job I have gotten on the slippery slope of micrography and hobby and my research blend into each other without me being able to draw a real line between them.

My Olympus BX61 is my main workhorse and I try to push it to the limits within my budget and time – most of my current projects are on fixed and embedded tissue sections, but recently I also dipped into live pondlife and am enjoying it very much. Observing the buggers is really satisfying and calming.

Macro work is also interesting and I did some optimization of normal handheld macro for documenting wet samples with a ringflash home modified for cross polarization with the Sony 90mm macro.

Inspired by what I saw on the forum I also want to explore supermacro in the near future. Also there is a box of Thorlabs components under my desk waiting to be unpacked…

I really enjoy reading the forums not only because there is much attention to technical details and ingenious solutions, but also to aesthetics and presentation – both of which are often underappreciate in scientific imaging (it’s just a tool for a job, after all). Amongst others I am very fond of the DIC photos posted by anne – pushing a Vanox to the limit, the technical excellent and visually pleasing autofluorescence images by WalterD and all the work done and presented by the other forum members. I also think that this forum is an excellent resource for technical and practical advice and creative ideas how to push the envelope of new and old equipment (my favorite example is the gradient universal filter by Litonotus).

For the future I hope to participate more actively in the exchange here and hope that within my scope I can provide expertise and I am looking forward to help and advice while setting up my supermacro setup.

Kind regards
Peter


Last edited by pbraub on Thu Dec 13, 2018 4:19 am; edited 1 time in total
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pbraub



Joined: 02 Feb 2018
Posts: 37

PostPosted: Thu Dec 13, 2018 4:17 am    Post subject: Reply with quote

Also some examples from recent work. This is more on the work than on the hobby side - but nevertheless taken on my setup:

UPlanFl 60XOI @ NA 1.25 with HR-DIC on a dry NA 0.9 condenser taken with a Point Grey GS2 monochrome camera in Micro-Manager.

The image is bronchial epithelium with a dual immunostain for different mucins (red and green) and a nuclear counterstain with DAPI (blue). The first image is a focus merge (ImageJ EDOF) of a 41 Image Z-Stack with 0.1 µm step. The second image a representative slice of the DIC channel from the center of the stack. The DIC image is flatfield corrected to remove the residual gradient.





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grgh



Joined: 09 Mar 2013
Posts: 188
Location: Lancashire. UK

PostPosted: Thu Dec 13, 2018 10:18 am    Post subject: as subject Reply with quote

Peter

Welcome to the forum, Looking at your WORK slide illustrations I feel sure that you will be a great asset to the group.

You will find lots of help and inspiration for your super macro goals.
Looking to see more of your voyage pictures in the future.

George
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and photography.
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ChrisR
Site Admin


Joined: 14 Mar 2009
Posts: 8233
Location: Near London, UK

PostPosted: Fri Dec 14, 2018 3:42 am    Post subject: Reply with quote

pbraub wrote:
Also some examples from recent work. This is more on the work than on the hobby side - but nevertheless taken on my setup:

UPlanFl 60XOI @ NA 1.25 with HR-DIC on a dry NA 0.9 condenser taken with a Point Grey GS2 monochrome camera in Micro-Manager.

The image is bronchial epithelium with a dual immunostain for different mucins (red and green) and a nuclear counterstain with DAPI (blue). The first image is a focus merge (ImageJ EDOF) of a 41 Image Z-Stack with 0.1 µm step. The second image a representative slice of the DIC channel from the center of the stack. The DIC image is flatfield corrected to remove the residual gradient.

If you post more images like that, I for one, will be absolutely delighted
How long do you get before the DAPI mage fades?
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pbraub



Joined: 02 Feb 2018
Posts: 37

PostPosted: Fri Dec 14, 2018 10:08 am    Post subject: Reply with quote

George, Chris, thank you for your kind words.

@ Chris
DAPI and fading is a non-issue for me. I am not sure if you mean fading over time (storage) or photobleaching during illumination.

The DAPI signal stays stable over months even at room temperature and i am sure that it will last even longer.
During illumination photobleaching is also hardly noticable.

We use Prolong antifade gold mounting medium suppemented with DAPI (https://www.thermofisher.com/order/catalog/product/P36931) under this condition also the immuno signal is fairly stable and you have to seriously abuse the slide to cause photobleaching even with a 100W mercury burner.
I recently switched to a LED source, with this I haven't noticed any bleaching at all.


Kind regards
Peter
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anne



Joined: 05 Dec 2014
Posts: 87

PostPosted: Sun Dec 30, 2018 8:05 am    Post subject: Reply with quote

Hallo Peter,
willkommen und vielen Dank für die Blumen!
Ich hoffe wir können hier bald mehr sehen von Dir.

Beste Grüße
anne
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