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Photo#93253
Setup for stacked images

Setup for stacked images
Licking County, Ohio, USA
January 20, 2007
Shown here is the setup I used for this palp photo, which was the result of 28 separate exposures combined with Helicon Focus Pro. I'm not 100% thrilled with this arrangement, but I'm getting decent results so I suppose it's worth sharing. I think I would get better (or at least more consistent) results by ditching the focusing rail and using the mechanical focusing stage from an otherwise broken microscope to raise and lower the photo subject.
  • Canon 30D
  • Canon Speedlite Transmitter ST-E2 (wireless flash controller)
  • Sigma 2X Teleconverter
  • Canon MP-E 65mm manual focus lens
  • Wimberley lens plate (barely visible here)
  • Really Right Stuff B150-B Macro Focusing Rail
  • Really Right Stuff BH-55 Pro ballhead
  • Gitzo 1325 tripod legs
Other equipment used but not shown here:
  • Two Canon 580EX Speedlite flash units
  • Quantum Turbo 2X2 Battery Pack
  • Canon RS-80N3 Remote Switch
Now for the good news: most of the equipment listed is optional for this task, which is great because you could buy a decent used car for what all this crap costs.
    You'll notice a few other items: black (now somewhat faded) construction paper, Post-it labeling tape (attached sticky side up), and a 35mm film canister with the bottom end cut off. The paper provides a little bit of light and glare attenuation. Post-it tape is used to hold the specimen in place without leaving a bunch of sticky goo to rinse off before returning to alcohol. And the film canister will be used as a softbox for the flash.

    Images of this individual: tag all
    Setup for stacked images Setup for stacked images Setup for stacked images Setup for stacked images

    focusing rail
    I've heard/read varying reviews on whether some focusing rails are OK for extreme macro, but your palp pic makes it look like this one can do very tiny incremental changes relatively easily and I just wanted to see whether you'd found anything better. Thanks a lot for this post btw.

     
    I don't recommend the rail
    Too sloppy for this work, and definitely not worth the money.

    I picked up an old microscope focusing stage off eBay for $20+shipping. Shown here.

    stacked images
    Thanks for posting this helpful how-to. Although I am unlikely to own a fine lens like the MPE 65 mm in the near future, it is very nice to see what it can do in the hands of a patient and precise user. The results are outstanding and certainly professional & publisable quality! Good work.

    may also
    want to consider remote control software - several of these out there, but this one seems to get good buzz.

    quite surprising
    not what I was expecting. Instead of a microscope focusing stage, you may want to consider a Lab Jack - I often use under my microscope for focusing.

     
    Alternatives
    I have a labjack that I occasionally use as an adjustable stage for photography, but this is for a single exposure. Helicon Focus software requires a set of images, each of which shows a small section of the specimen in focus. These are taken in sequence, starting with the most distant part of the subject in focus, and in the case of the palp, increasing the distance in increments of about 0.02 mm. until the closest part of the subject is in focus. The 28 exposures with increments of 0.02 mm., would cover a specimen front to back thickness of slightly over 0.5 mm. The software then assembles the sharp part of each exposure into the final image. These numbers are my estimates, based on the size of the specimen and the number of images taken.
    I don't think my Labjack, or any other Labjack that I am aware of, is capable of this kind of precision.
    Gayle

     
    Is it critical
    as to the sequence of exposures? Should one start at the closest surface of the subject and move the camera downwards or start at the furthest part of the subject and move the camera upwards; or is it unimportant?

     
    Sort of...
    Helicon Focus likes to start with the closest image first and then work its way down, but you can take the exposures in any sequence and just change the sort order within the software if needed. I start shooting with the furthest part just because I get smoother action when drawing the camera upward rather than downward.

    The latest version of Helicon Focus is supposed to be able to automatically determine the ordering but I disabled that option since it seem to work quite right for me.

    If you've worked with AutoMontage the concepts are pretty similar. Helicon Focus is a little easier for me to use and seems to have a much greater tolerance for images in the stack that aren't lined up properly either by placement, rotation, magnification and even brightness to some extent. In other words, it will detect those issues and do a pretty good job of recovering from them. This image looked like a trainwreck with AutoMontage. With Helicon Focus it took about 10-15 minutes of clean-up work.

     
    Thanks Jay
    I've downloaded the program and will try it with some tabanid heads.
    I work in a horizontal plane with the camera and lens locked onto a RRS Camera Bar; I use focussing rails to move the subject in 3 dimensions. I guess this means I should focus on the front of the subject and move it forwards towards the lens.
    Your images are superb and have inspired me to do better with my photography.

     
    Thanks Tony
    That's quite a compliment. Thank you.

    Your photos already look great, so I look forward to seeing what you come up with. It's a relief being able to open the aperture for additional sharpness without sacrificing depth of field. (photos for the palp I linked to above were shot at f/4)

    BTW, I left an entire word out of my previous response. Second paragraph should have said "since it doesn't seem to work quite right for me."

     
    How
    did you do that? Your 'this image' looks like a live specimen - how did you get it to sit still for that? I've used AutoMontage a couple of times and it seemed to be quite sensitive to varying depths - steps had to be quite evenly spaced.

     
    Liquid nitrogen :-D
    Yes, it was a living specimen. In fact he's still living today. He was in that pose for about 30-40 seconds for that image. After I got a few exposures with no movement I quickly opened the aperture to f/11 and started taking a series for stacking.

    I'm afraid I have no idea how to get a living jumper to sit perfectly still for any length of time though. Having photographed 200+ jumpers in the last couple years, some of them five or six times through different molts or just to get a specific angle, I can say that it doesn't happen often. Phidippus princeps does seem to be pretty calm most of the time, though the females almost constantly drum their palps onto whatever they're standing on.

    A long lens (=>180mm) is an absolute requirement for shooting jumpers IMO, and calmly easing them into the 'studio' area from their container can be a big help as well.

     
    high precision labjacks
    the type of labjack I had in mind is the kind used for optical/laser benchwork [thorlab,newport,etc] - they have a very high degree of precision - capable of µM adjustment. I occasionally see them on e-bay for bargain prices - new they're VERY pricey. I like them because they are heavy, stable and repeatable. The LJ750 Compact Lab Jack
    looks promising.

     
    moving camera vs moving subject
    Quite awhile back, Syncroscopy, the Automontage company, used to have a motorized stage type of thing to move the subject, but now they tout a motorized scope head to move the "camera". I've used both and the problem with moving the subject for image-stacking is that the lighting changes for different "slices". It doesn't matter as much at higher magnifications, but can matter a lot at lower ones, esp with very convex subjects.

     
    True...
    Keep in mind much depends on the type of lighting used, and how it is applied. With a moving subject, using a soft, diffuse light source will give less of a light-change between slices than a hard, point-source light. And if the light source is also moving with the subject, then you should notice almost no changes between slices.

    I agree it's best to move the camera between slices, but the camera is a big heavy beast compared to the subject being photographed so it's nice to have options.

     
    Thanks Phillip,
    The LJ750 appears to be the ideal device for use with the Helicon Focus software. The specs are amazing. If I don't see one on e-bay before warm weather sets in, I might have to spring for a new one.
    Thanks again,
    Gayle

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