Stacked pictures come out Cartoonish

Do you have any microscopy questions, which you are afraid to ask? This is your place.
Post Reply
Message
Author
Plasmid
Posts: 566
Joined: Mon Sep 28, 2020 5:34 am
Location: North GA
Contact:

Stacked pictures come out Cartoonish

#1 Post by Plasmid » Mon Dec 14, 2020 5:30 pm

I keep getting this cartoonish effect when stacking pictures, no more than 5-7 at the time using CombineZP, is this a program related issue?
Attachments
Diatom colony.jpg
Diatom colony.jpg (108.77 KiB) Viewed 6278 times
Copepod.jpg
Copepod.jpg (200.68 KiB) Viewed 6279 times
Algae on Algae.jpg
Algae on Algae.jpg (95.35 KiB) Viewed 6279 times
Algae on a grain of sand.jpg
Algae on a grain of sand.jpg (209.43 KiB) Viewed 6279 times

MicroBob
Posts: 3154
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Stacked pictures come out Cartoonish

#2 Post by MicroBob » Mon Dec 14, 2020 6:25 pm

Can you post a single image for comparison?
As far as I remember you use a USB camera - right? These often don't offer the very best raw image quaöity due to small and low end sensors and counter this with heavy internal image corrections. That is not the best starting point for image stacking.

Plasmid
Posts: 566
Joined: Mon Sep 28, 2020 5:34 am
Location: North GA
Contact:

Re: Stacked pictures come out Cartoonish

#3 Post by Plasmid » Mon Dec 14, 2020 6:30 pm

one of the pics used, yep the USB cam does not offer a lot of quality.
Attachments
sample.jpg
sample.jpg (15.22 KiB) Viewed 6267 times

BramHuntingNematodes
Posts: 1538
Joined: Tue Jan 21, 2020 1:29 am
Location: Georgia, USA

Re: Stacked pictures come out Cartoonish

#4 Post by BramHuntingNematodes » Mon Dec 14, 2020 7:03 pm

I thought that's what stacking was supposed to do
1942 Bausch and Lomb Series T Dynoptic, Custom Illumination

MicroBob
Posts: 3154
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Stacked pictures come out Cartoonish

#5 Post by MicroBob » Mon Dec 14, 2020 9:50 pm

Plasmid wrote:
Mon Dec 14, 2020 6:30 pm
does not offer a lot of quality.
Well, at least the images are tiny! 8-)

I'm no big stacking expert but what suprised me was that stacking doesn't necessarily lead to better images. In case of too few images I got stange looking and unsharp results. In case of objects with contrasty structures in multiple planes it doesn't give a natural image when all structures are combined in one image.
As far as I know combine ZP has not been updaten for ages. You might give PICOLAY a try, especially when you are prepared to read the manual.

Most people use stacking for dependably static subjects. Maybe it would be better to try this out with such an object to exclude errors due to movement.

Bob

SWmicro
Posts: 134
Joined: Sat Nov 07, 2020 10:52 pm
Location: England.

Re: Stacked pictures come out Cartoonish

#6 Post by SWmicro » Mon Dec 14, 2020 10:05 pm

Plasmid wrote:
Mon Dec 14, 2020 5:30 pm
using CombineZP, is this a program related issue?
Do you mean related specifically to ZP or stacking in general ?

I was going to say much the same as MB but I went away to google ZP to see if it had been recently updated or adopted by some other team, so he beat me to it :) !

I used to use Combinz Z5,ZM & ZP but it is unsupported and has a curious user interface - I now use Picolay. It works well (usually even straight out of the box on default settings for a quick try)
It does not need installing, it can be used from a USBstick or other folder/directory on HD. It is free and maintained frequently.
Give it a try and see if your cartoons persist ?
Last edited by SWmicro on Tue Dec 15, 2020 12:01 am, edited 1 time in total.

User avatar
micro
Posts: 203
Joined: Mon Jul 22, 2019 6:11 am

Re: Stacked pictures come out Cartoonish

#7 Post by micro » Mon Dec 14, 2020 11:45 pm

I have the same issue a lot of the time. Stacking seems to create big thick lines making the subject look like it was outlined with a pencil. I think its just an issue with microscope resolution and such and maybe photoshop not really getting how to deal with microscope images or something. I don't think photoshop knows which shadows to remove and which to keep. Only solution I can think of is to divide up parts of an image and stack each piece individually so the shadows are where you want them and then paste them together.

MichaelG.
Posts: 3976
Joined: Mon Apr 10, 2017 8:24 am
Location: North Wales

Re: Stacked pictures come out Cartoonish

#8 Post by MichaelG. » Tue Dec 15, 2020 12:46 am

Plasmid wrote:
Mon Dec 14, 2020 6:30 pm
one of the pics used, yep the USB cam does not offer a lot of quality.
.

Sorry, but I don’t think there is any detail in that image which is sharp enough to merit stacking.
You need source images which are ‘tack sharp’ in the plane of focus and unsharp elsewhere.
... the stacking software selects and combines the sharp regions from each of them.
It will do the best it can, but low resolution jpg files are not a good source. :(

MichaelG.
Too many 'projects'

einman
Posts: 1508
Joined: Tue Feb 24, 2015 1:03 am

Re: Stacked pictures come out Cartoonish

#9 Post by einman » Tue Dec 15, 2020 2:55 am

Stacking only improves the image if the total depth of your subject can be divided into "slices" or multiple images. Each "layer" sharply focused on a given plane, then combined to make one sharper image of a much deeper depth than you can achieve with a single image. You must know what each turn of your fine focus yields in terms of depth. If, for example the smallest increment (vertical movement) you can achieve on your fine focus is say 10 microns and your specimen is only 5 microns in depth stacking does absolutely nothing for the specimen but will improve background imaging. Why? You start with a focused image on a given plane. You adjust the fine focus, moving the focus 10 microns and are now focusing past the specimen. Take as many images as you want, it will improve nothing. I am amazed at individuals that take literally dozens of images where their fine focus easily exceeds the depth of the specimen with one or 2 miniscule turns. They could have achieved the same or better results with perhaps 2 or 3 images. If you take multiple images in that type of situation you increase your chances of introducing errors in the image as well. This is especially true if the subject is moving however slightly. Although you can account for some movement in the better programs. The best stacked images are achieved by individuals that understand the specifications of both the objective being used, in terms of resolution and DOF, as well as the capability of their scope in terms of the focus mechanism.

For example 1 division on the fine focus of my Leitz Diaplan is about 2 microns per the manual. This of course assumes the focus mechanism is not worn etc. So If I were trying to stack images of say a Rotifer it would take most likely no more than 4 images covering a depth of about 8 microns, assuming it was lying flat and not positioned at an angle etc. Often you can test the depth before actually taking pictures. You start by just bringing the image into focus. Then you adjust using the smallest increment of focus your scope allows, until you see additional features and keep doing this until the image goes out of focus, counting each incremental turn of the focus knob. That final number is the number of stacks you should take.

BramHuntingNematodes
Posts: 1538
Joined: Tue Jan 21, 2020 1:29 am
Location: Georgia, USA

Re: Stacked pictures come out Cartoonish

#10 Post by BramHuntingNematodes » Tue Dec 15, 2020 3:04 am

Also in my experiments I find that too many shots of a thick specimen can lend a strange almost cubist abstraction as the program tries to make an image of both the front and back of an object.
1942 Bausch and Lomb Series T Dynoptic, Custom Illumination

einman
Posts: 1508
Joined: Tue Feb 24, 2015 1:03 am

Re: Stacked pictures come out Cartoonish

#11 Post by einman » Wed Dec 16, 2020 2:22 am

I must complement Plasmid. At least he is taking a reasonable number of shots. Most likely he just needs to refine his technique a bit.

hans
Posts: 986
Joined: Thu May 28, 2020 11:10 pm
Location: Southern California

Re: Stacked pictures come out Cartoonish

#12 Post by hans » Wed Dec 16, 2020 3:31 am

Plasmid wrote:
Mon Dec 14, 2020 6:30 pm
one of the pics used, yep the USB cam does not offer a lot of quality.
It looks like the USB camera software is clipping blacks, is there a contrast setting anywhere? Not familiar with stacking, but possibly turning the camera contrast down to something more neutral then adjusting after stacking if necessary might give better results? Spike at left side of histogram:
hist.png
hist.png (11.19 KiB) Viewed 6122 times

User avatar
75RR
Posts: 8207
Joined: Sun Oct 12, 2014 2:34 am
Location: Estepona, Spain

Re: Stacked pictures come out Cartoonish

#13 Post by 75RR » Wed Dec 16, 2020 11:57 am

.
What stacking does, by combining images at different focus points, is effectively extend the depth of field of a given objective.

There is no reason why that by and of itself should give a better image - it just gives an image with a greater depth of field.

Better images come from good subjects, good illumination technique, good objectives and good operator skills.
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

User avatar
cuxlander
Posts: 146
Joined: Sat Sep 17, 2016 12:13 pm

Re: Stacked pictures come out Cartoonish

#14 Post by cuxlander » Wed Dec 16, 2020 2:23 pm

Hello plasmid,
one of the pics used ...
I feel that the single source image already shows sharp edges.
Rgds,
Hans

User avatar
Roldorf
Posts: 574
Joined: Tue Apr 23, 2019 6:58 pm
Location: Northern Germany

Re: Stacked pictures come out Cartoonish

#15 Post by Roldorf » Thu Dec 17, 2020 9:53 am

Make sure you don't stack images from below the halfway point of the subject and that you have the correct order of images, either from the top down or bottom up (middle) (depending on which order you took the images) in the settings.
Location: Northern Germany

Unknown Brand: Optika SFX 91: Bresser Science Infinity: Canon 4000d
ImageImage

EYE C U
Posts: 288
Joined: Wed Jul 15, 2020 5:18 pm

Re: Stacked pictures come out Cartoonish

#16 Post by EYE C U » Thu Dec 17, 2020 12:36 pm

I DON'T KNOW ABOUT Z BUT IN HELICON FOCUS YOU CAN EDIT OUT THOSE REMNANTS BY BRUSHING OVER THE BEST FRAME FOR THAT LAYER GIVING YOU A CLEAN EDGE

Post Reply