Cyclops (24 image stack)

Charles Krebs · Post by **Charles Krebs** » Sat Apr 02, 2005 10:34 pm

A while ago in the "techniques" section we discussed combining images to gain depth-of field. I had mentioned my search for software that could help automate the process, and was very pleased with Helicon Focus. This type of picture is precisely the reason I was on that quest. This is a cyclops (a ventral view) taken with a 10X objective. It's just not possible to record all the fine detail in a single exposure because the subject is so "deep" and DOF so shallow. It would have been a nightmare to try to do this in Photoshop (especially the apendages at the top of the image!) . But it took less than minute to combine 24 images with the program, and "clean-up" was pretty minimal.

Kenv · Post by **Kenv** » Sun Apr 03, 2005 12:29 am

Thats astonishing Charlie but I'm already one of the converted.
I don't often come across these things, hopefully that will improve.
Kenv

lacerta · Post by **lacerta** » Sun Apr 03, 2005 12:09 pm

Very nice Charles. The image is reminiscent of Victorian era botanical print. Almost like a detailed pen and ink with watercolor wash.
George

Charles Krebs · Post by **Charles Krebs** » Sun Apr 03, 2005 2:09 pm

I thought it would be fun to look at one image of the 24 that were used, and compare it to the final composite. I marked a few spots (arrows) that show what pieces of this image were used in the final image. It's pretty obvious how tough it would be to show the upper appendages clearly with one picture.

Kenv · Post by **Kenv** » Sun Apr 03, 2005 3:39 pm

Charlie, I have to assume that the cyclops was dead, or didn't move. I know that H.F. can resize the image and possibly change the view - I did one stack in which the resultant image showed the edges of the other images over-lapping, a bit like a displaced stack of cards.
Kenv

S. Alden · Post by **S. Alden** » Sun Apr 03, 2005 4:17 pm

This is really nice and I decided to d/l the software, but when I went to install it, I received an "exception error". I d/l again and the same thing, so I suppose I cannot have this software.

In any case, software or not, it takes talent to get an image with this quality.

psneeley · Post by **psneeley** » Sun Apr 03, 2005 6:28 pm

I suppose the time is not too far distant when the scope will automatically and electronically focus through various levels of the specimen and produce a stacked image to the screen (or eyepiece) that banishes entirely the DOF problem forever -- the tyranny of DOF vs high N.A. will be gone . . . all in a matter of microseconds. I look forward to that day because it will open up the microworld even more. Hartley, in his book, notes that early microscopists had to 'fight' for an image so that they could produce the wonderful and detailed drawings for their books . . . I fight, sometimes, at the higher powers, just to figure out what in the heck I'm looking at

But what you've produced here is just amazing -- Helicon focus seems like a great tool.

Kenv · Post by **Kenv** » Sun Apr 03, 2005 6:32 pm

Hi Sue, Its takes talent and very good equipment to do as good as Charlie but that is no reason for you not to use Helicon Focus, its a brilliant program and its dead easy for anyone to use and you will get good results from it. I'm certain that there is no reason why you shouldn't be able to download it, you may need to change a setting in your computer and I'm sorry but thats not my forte so I can't help - I'm sure someone can though.
Kenv

S. Alden · Post by **S. Alden** » Sun Apr 03, 2005 6:38 pm

Kenv wrote:Hi Sue, Its takes talent and very good equipment to do as good as Charlie but that is no reason for you not to use Helicon Focus, its a brilliant program and its dead easy for anyone to use and you will get good results from it. I'm certain that there is no reason why you shouldn't be able to download it, you may need to change a setting in your computer and I'm sorry but thats not my forte so I can't help - I'm sure someone can though.
Kenv

Thanks KenV. I can download the software, I just cannot install in on my computer. I get that "exception error"

Charles Krebs · Post by **Charles Krebs** » Sun Apr 03, 2005 8:57 pm

Steve... that day is here already. At least one company makes a program (AutoMontage) that can work with a motorized "Z" control (focus). It will calculate how many shots are needed in the stack, control the motorized "Z" control, and take the photo at each step. It can also take a bracketed series of shots at each step to increase dynamic range (avoid washed out highlights and detail-less shadows) and then combine the stack and exposures automatically. I don't know how well it works and probably never will .......$$$$$$......

KenV... this subject was actually alive and stayed still. I did have quite a few "stacks" that I had to abandon after it moved a little. Living subjects are tough. Your subject really can't move at all, or the program thinks the detail in the "shifted" shot is needed.

Frez · Post by **Frez** » Mon Apr 04, 2005 6:06 am

Great work Charlie! The symmetry is amazing even down to the hairs on the legs. Did you adjust any parameters in Helicon or are they all at their default settings?

Frez

piotr · Post by **piotr** » Mon Apr 04, 2005 3:24 pm

Fabulous picture, Charlie. I agree with George - it looks like a drawing. Amazing!

piotr · Post by **piotr** » Mon Apr 04, 2005 4:00 pm

By the way - on the subject of 'freezing' cyclops. One can use magnesium sulfate (Epsom salt) to anesthetise these little crustaceans (may work with other specimens, as well). Just add a drop of concentrated solution of magnesium sulfate to a drop where your specimen is. It should stop moving. The best part - this method is reversible. The animal will 'wake up' after putting it back to water.

twebster · Post by **twebster** » Wed Apr 20, 2005 10:16 am

Hi ya' Charlie

You've just been creating some incredible images with Helicon Focus. Your work is an inspiration to us all. It's only natural that this image was chosen for the Administrator's Appreciation Gallery and the front cover of www.amateurmicroscopy.net.

Thanks for your continued participation, my friend

Sven Bernert · Post by **Sven Bernert** » Wed Apr 20, 2005 1:50 pm

WOW Charles! This is really something! Congrats to one unbelievable image.

Best,
Sven