Best setup for Stemi 2000C photography?

Begonnen von Heterop, Oktober 09, 2021, 22:51:27 NACHMITTAGS

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Heterop

Hi,
This is my first post in this forum, please excuse me for the English (:
I am looking for an advice on how to improve the quality of my microscopic photography. My setup includes Stemi 2000C with a Canon Powershot G5 attached to Camera Adapter (# 426126) And KL1500 LCD as a light source.

To demonstrate the problem I manually stacked 9 images at a maximum magnification of x5 and a maximum zoom at G5.
As you can see there is a lot of noise and the image is not sharp enough. How do you think I can improve the quality? Replace the G5 with a newer model, say G11? Use another lighting source, maybe use a flash instead? ...

Kind regards
Hetero

knipser009

#1
Hi

In my opinion the famous Zeiss 2000c is not suitable for making such stacks as you did. The focus seams to be in a part of the eye of the insekt and does not take the total  depth of the insekt.
9 pictures to get a good stack are not enough.  I think 100-150 pics will be a good choise. 
Viele Grüße aus dem SaarPfalzKreis

Wolfgang
gerne per "Du"

Peter V.

#2
Hello,

do you understand german language? If the answer ist "yes" we can proceed the discussion in German, otherwise in English. If you can only read German, we can answer in German, nevertherless you can write in English.

Indeed a normal stereo microscope without a special device for stacking is not a good tool for focus stacking but for me it is not the reason for your noisy image which gives an impression like looking through a wall of fog. Can you please show one of the single images of the object through your microscope?

As you see below it is possible to improve your image without great efforts. First I removed the  "foggy curtain" with ImageJ and the Plugin "CLAHE" (both programs are freeware) I think, CLAHE is the best tool for this. Later I denoised in Photoshop Elements with the Plugin "NeatImage", after that I enhanced sharpness and contrast in PS Elements. Thats all.

Regards Peter

PS: Please write a short introduction to your person in "Mikroskopiker im Netz". I prefer discussions with members with a first name, hetero or homo does not matter  ;)

Dieses Post wurde CO2-neutral erstellt und ist vegan. Für 100 Posts lasse ich ein Gänseblümchen in Ecuador pflanzen.

Heterop

#3
@knipser009 stack of 150 images is hard to do manually. For stacking I just turn the focus knob and take click on camera shutter. Is it is possible to attach automatic focus rail to this microscope with Canon Powershot G5?

@Peter thanks for tips. I usually fix the image with Photoshop. but the image quality is still very far from the sharp and natural color images I can see here in the forum. I wonder if I can improve anything is the setup of the microscope to get a better images (before editing).
The details for the single image are f/3 stop, exposure 1/8 sec. this is automatically selected by the G5 when I select manual photo with ISO 50 to reduce noise.

I attached example of image after editing with Photoshop and a single image from my original 9 photo stack (I can't upload the original image since it's over 1024k).

P.S. I don't read German very well but I can use translation tools when needed to, so you can answer as you wish.

Heterop

#4
deleted

Peter Reil

Zitat von: Hetero in Oktober 11, 2021, 14:06:50 NACHMITTAGS
but the image quality is still very far from the sharp and natural color images I can see here in the forum.

Hallo Hetero,

das ist normal. Die Spitzenfotos, die du hier sehen kannst werden von Personen gemacht, die viel Erfahrung und Übung mit ihrem Equipment haben.
Das kann nicht jeder - ich auch nicht.

Ich empfehle: Üben, üben, üben,.......

Gruß
Peter

Meine Arbeitsgeräte: Olympus BHS, Olympus CHK, Olympus SZ 30

Lupus

Hello,

maybe you should reduce the stray light from the microscope objective as a first step. Normally, you do not use a field diaphragm with stereo microscopes. But you can reduce the stray light by using black paper/cardboard as a background and also illuminate only the image field if possible. And you can use a white background - if desired - with a small diameter adapted to the image field.
Stacking also works better if the source photos have a higher contrast.

Hubert

witweb

#7
Hello Hetero,

In addition to all the hints that the other users have already given, here is a tip for editing the images: https://www.mikroskopie-forum.de/index.php?topic=42168.msg310854#msg310854

BR

Michael
Leitz Orthoplan
Zeiss Standard 18 mit Fluoreszenz-Auflichtkondensor IV FL
Lomo Biolam, Motic SMZ-168
Canon EOS 750D
https://mikrokristalle.net
https://www.youtube.com/@Mikrokristalle

Lupus

Hallo,

man braucht nicht unbedingt kostenpflichtige Bildbearbeitungsprogramme, um solche Bilder zu verbessern. Fast jede unkomplizierte Freeware lässt das zu. Als Beispiel links das kontrastlose Originalbild, das eingeblendete Histogramm zeigt den schlecht ausgenutzten Kontrastumfang.

In der Mitte das Ergebnis nachdem die Schatten und Lichter manuell innerhalb des maximalen Belichtungsspielraumes mit einem einfachen Bildbearbeitungsprogramm gestreckt wurden (zusätzlich wurde in den Schatten der Farbstich durch manuelles Verschieben zwischen den drei Farbkanälen etwas korrigiert).

Rechts die von Michael beschriebene automatische Photoshop-Optimierung. Man erkennt am Histogramm dass dort zusätzlich der Kontrast angehoben wurde, speziell in den dunklen Bereichen (Überhöhung der Schatten-Peaks links), gut auch sichtbar an der Betonung der dunklen Haare des Insekts. Das kann man aber mit fast jedem einfachen Bildbearbeitungsprogramm in einem zweiten Schritt auch erreichen. Das Photoshop-Bild zeigt bei höherer Vergrößerung bereits deutliche Artefakte und Verstärkung des Bildrauschens.

Hubert

Heterop

Thanks all for the good advice,
My only concert is that by increasing the contrast it also change the colors so they become more vivid and less natural. It looks great for artistic purpose but what do you do if you want to use the image for scientific purposes?
Is there a way to remove the fog from the image and keep the colors as natural as possible, meaning that the hue will look exactly as you see it when you look through the eyepiece under a microscope?

Peter V.

Hello,

what exactly is the "real" colour of the object?  ;) It is a great difference between lighthing with daylight, halogen or LED. Furthermore the colour of an image depends on the firmware of the camera. A picture taken with my smartphone camera has a different colour saturation than a picture taken with my DSLR. In my opinion "objevtive" colour or an "objective" image does not exist.

Regards
Peter
Dieses Post wurde CO2-neutral erstellt und ist vegan. Für 100 Posts lasse ich ein Gänseblümchen in Ecuador pflanzen.

Heterop

Zitat von: Peter V. in Oktober 12, 2021, 18:36:49 NACHMITTAGS
Hello,

what exactly is the "real" colour of the object?  ;) It is a great difference between lighthing with daylight, halogen or LED. Furthermore the colour of an image depends on the firmware of the camera. A picture taken with my smartphone camera has a different colour saturation than a picture taken with my DSLR. In my opinion "objevtive" colour or an "objective" image does not exist.

Regards
Peter

Yes you are right, you could say that "true color" is a matter for philosophical discussion. On one hand the color is interpreted by the camera and on the other hand the color is interpreted by the screen that displays it. So to make it a more mundane question in my opinion the original photo with the fog show the closest color to the image thought the eyepiece. Is it possible to remove the fog without changing the saturation of the colors?

Regards.

Lupus

#12
Zitatin my opinion the original photo with the fog show the closest color to the image thought the eyepiece
This does not seem plausible. The colour saturation reduces at the same time as the contrast decreases, e.g. due to stray light. But regardless of this, you can easily change the colour saturation independently and reduce it again by hand.

ZitatIs there a way to remove the fog from the image and keep the colors as natural as possible
In photography, one usually uses a grey card for white balance and colour cards for colour matching. You can do this similarly with a stereo microscope, e.g. by photographing small pieces of coloured areas under the same conditions as the object and transferring the colour and contrast adjustment of the image processing programme to the object photography.

Hubert

mete

Zitat von: Lupus in Oktober 12, 2021, 19:39:56 NACHMITTAGS
In photography, one usually uses a grey card for white balance and colour cards for colour matching. You can do this similarly with a stereo microscope, e.g. by photographing small pieces of coloured areas under the same conditions as the object and transferring the colour and contrast adjustment of the image processing programme to the object photography.

I use color targets for normal/macro photographs and they are pretty straightforward to use but I havent seen a color target for transmitted light microscopy. Do you know any such product, eg a color target microscopy slide ?

Mete

Lupus

Zitat... but I havent seen a color target for transmitted light microscopy. Do you know any such product, eg a color target microscopy slide ?

For stereo microscopes (normally in reflected light), one does not need a transmitted light test image. I also don't know of any special transmitted-light slides for this purpose. In transmitted light microscopy - e.g. with stained samples - I don't see much sense in it either. You can do the grey balance with the background illumination. But there are colour test slides for 35 mm photography. You could possibly use them for microscopy as well. You can also match individual colours with thin foil colour filters.

http://fotowand.de/frameset.htm?CHOICE=/alink.htm&SHOW=/fwtc-mai.shtml
http://fotowand.de/8845.htm
http://fotowand.de/9964.htm

Hubert