My first real micro challenge

hotrodder19 · Post by **hotrodder19** » Tue Jul 09, 2019 6:49 pm

Ok Guys, just take it slowly with me re this result. Using my trusty D750, 50mm lens and BE Plan 10x micro. Have attached the Pmax Zerene output pic , the result of some 1200 images ( yes a lot of time). Also attached are a sample of my images taken that formed part of the overall pic for your assessment. Yes I need some work but first I naturally need to know what I did wrong and how to change things up. I await the Admin's humble comment and advice. The last pic is the Pmax zerene one.

Cheers hotrodder19

rjlittlefield · Post by **rjlittlefield** » Tue Jul 09, 2019 7:32 pm

Using my trusty D750, 50mm lens and BE Plan 10x micro.

50mm? Why that lens, instead of the 200mm that we've talked about before?

--Rik

hotrodder19 · Post by **hotrodder19** » Tue Jul 09, 2019 7:50 pm

Hi Rik, just thought I'd try and make it easier for myself first up .--not a good choice ?

rjlittlefield · Post by **rjlittlefield** » Tue Jul 09, 2019 9:07 pm

My engineer friends used to joke among themselves, "If you can't get something to work, read the instructions. If you still can't get it to work, follow them."

Despite appearances, this was not intended to be insulting and it was not totally a joke. Common knowledge is that engineers often read instructions the first time mostly for insight. If the resulting insight is enough to get the thing working, great. But if the insight from the first reading (or second, third, whatever) is not enough, then it's time to simply follow the instructions, step by step, in hopes that the process will work correctly even though the person doing it does not yet understand exactly what's important and what is not.

So, without intending to be insulting, I will now tell you bluntly that you'll get much farther much faster if you actually follow the instructions.

[50mm] not a good choice ?

Correct, using a 50mm lens was a bad choice for a number of reasons that I will not take time to try explaining. Likewise for shooting 1200 frames. That was almost entirely a waste of time.

Instead, follow these instructions:

Use your 200 mm lens, focused at infinity, set wide open, with no extensions between the 200 mm and the BE Plan 10x.
Light and frame your subject.
Screw the focus rail backward until the whole subject is out of focus.
Screw the rail forward until something on the subject definitely is in focus. I suggest starting with a part of one eye in focus.
Shoot a stack of only 50 frames, continuing to screw the rail forward, using a focus step of 0.01 mm or 0.0004 inches, whichever is convenient on your micrometer. Note that the total depth of this stack will be only about 1/2 mm, since 0.01*50 = 0.5. For this exercise, you will not be getting the entire face in focus.
Run the resulting 50 frames through Zerene Stacker PMax.
Show us the first frame, the last frame, and the processed result, simply resized to fit forum limits.
Show us also an "actual pixels crop" from the sharpest part of one of the source images. See the FAQ: What is '100% crop' or 'actual pixels crop' to be sure what I mean by "actual pixels".

--Rik

hotrodder19 · Post by **hotrodder19** » Wed Jul 10, 2019 1:52 am

Well Rik a crushing start to microphotography. Realistically I am and I know other beginners who really never knew or saw any instructions on how to do photo stacking using microscope lenses apart from bits picked up on this site. If there were detailed instructions somewhere on the site then they need to be made more accessible for beginners . My understanding from what I have seen had me going left to right across the insect head producing the end result, but it seems I need to go from far out to close in one AP plane, is this correct ? I must admit that one of my next questions was going to be how to cover the various changes in an insect for example from body to head etc and maybe legs which are all on differing planes. So all pics in microphotography are AP forward back moves with the lens ? I await your comment on this. Have done the new stack as requested and will list them in the order you requested, ie 1st pic, last pic, Pmax and then the pixel count ok.

Not easy with sizing and appropriate style but hopefully you can open them all.

Cheers Russell hotrodder19

Adalbert · Post by **Adalbert** » Wed Jul 10, 2019 2:40 am

Hello hotrodder19,
As far as I can see you are stacking with AF of the photo-lens, aren’t you?
So, I have already taken many photographs with the BE Plan 10x and BE Plan 4x.
You can stack with AF and use BE Plan 4x but you cannot stack with AF and use the BE Plan 10x.
The smallest step of the AF-motor is too big for the DOF of the BE Plan 10x.
So, the stacked results are not really sharp, because the stacking-steps are too big

But the BE Plan 4x works perfectly :-)
If you use 200mm (e.g. CANON EF 70-200L) photo-lens then you will get the magnification about 4x.
If you use the macro-lens 100mm (e.g. CANON EF 100L macro) then you will get 2x.
BR, ADi

hotrodder19 · Post by **hotrodder19** » Wed Jul 10, 2019 6:06 am

Adalbert, where did you come from to analyse and critique my first offering ?? At first I though what is this guy on about with AF etc etc .............however well I thought I'd better check to make sure it wasn't accidently switched to AF, because at a point it was on manual , MMMmmm ! Well yes it was on AF , can't say how it got back there from manual but it was, so i will start again and send Mr Rik some new pictures.

Thanks Adalbert. Cheers hotrodder19

hotrodder19 · Post by **hotrodder19** » Wed Jul 10, 2019 6:07 am

Adalbert, how did you deduce that the camera was set to AF ??

hotrodder19

hotrodder19 · Post by **hotrodder19** » Wed Jul 10, 2019 7:31 pm

Ok, Rik and Adalbert, have re done the pics as requested this time ensuring setting was on M in camera--sorry. Will list in order Rik requested.

Ok done, now for comment. Thanks hotrodder19

rjlittlefield · Post by **rjlittlefield** » Wed Jul 10, 2019 9:04 pm

This is looking better. Now the whole frame is filled with image, there are no gross misalignment problems, and the stacked output file has hugely more DOF than the originals, with no obvious focus banding.

The big problem now is that it looks like you're getting quite bad motion blurs, predominantly in the vertical direction.

Typically this is caused by mirror and/or shutter motion, in combination with continuous illumination.

As mentioned in earlier discussions, the most reliable cure for this is to expose by electronic flash, preferably with the flash unit manually turned down to something like 1/16 power so as to get effective exposure times around 1/5000 second. Mirror lockup is always a good idea, with several seconds delay between mirror up and actual exposure.

Even better (but probably overkill in this case) is to shoot in a darkened room, using flash exposure, with mirror lockup plus a couple of seconds of shutter open time, with flash trigger on the second curtain. That way you get the vibration-freezing effects of flash, plus a couple of seconds delay between the shock of shutter-open and the flash that does the actual exposure.

If you can't shoot with flash, then at the very least you need to use mirror lockup.

Just for the record, what sort of illumination, shutter time, and mirror lockup delay did you use for these images?

--Rik

hotrodder19 · Post by **hotrodder19** » Wed Jul 10, 2019 9:44 pm

Hi Rik, I used 125 @ 6.3.. I used live view, delay was approx 5 sec. I used wifi thru tablet to take photos. With what I have done so far, how do I get the whole head in my picture ? You would have seen my comment yesterday re my understanding of photo stacking and going from left to right etc, can you enlighten me on how to go the next step to get a "whole" image in focus . Is this left to right wrong ?

Look forward to your comments, hotrodder19

ChrisR · Post by **ChrisR** » Thu Jul 11, 2019 2:47 am

You need to use the 200m lens wide open, or you will get vignetting.
Vibration will be difficult or impossible to avoid unless you use flash - are you?
"Left to right" isn't clear - furthest to closest is the better direction but it doesn't matter as long as you have all the picture you want, at both ends.

hotrodder19 · Post by **hotrodder19** » Thu Jul 11, 2019 5:49 am

Thanks Chris R, with respect to the 200mm lens are you referring to it being at infinity ? You say wide open, is this the same ? These pics were taken with the lens at infinity. As an add on here I meant to tell Rik that in respect to lighting i used 3 Jansjo mini lights like he suggested plus I had 2 surround 15w LED around just increasing overall light in the area.

rjlittlefield · Post by **rjlittlefield** » Thu Jul 11, 2019 7:58 am

"Wide open" always means to set the lens at its smallest f-number, with its aperture open as far as it will go. According to your picture at https://www.photomacrography.net/forum/ ... 343#251343, that will be f/4 for your 200 mm telephoto. This has nothing to do with focus.

how do I get the whole head in my picture ?

The simplest way is to use a different lens, one that is appropriate for that job. That 10X CFI BE Plan objective is not appropriate to shoot the whole head. Nikon rates it as being designed to cover an area that is only 1.8 mm in diameter. ("OFN 18" means an 18 mm diameter image at 10X, then 18/10=1.8 mm at subject.) As a matter of practice the objective will cover a larger area, 2.8 mm frame diagonal in your current case. With great care, some good luck, and a different rear lens, that objective can be pushed to cover about 5.6 mm frame diagonal, albeit with some corner darkening. But that's not something that a beginner would have success with. Alternately, there's a complicated and tricky process called "stack-and-stitch", described at https://www.photomacrography.net/forum/ ... php?t=9542 . Stack-and-stitch is not something a beginner should be messing with either.

At this point you should be concentrating on making a good picture of the small area that the objective is intended to cover.

I used 125 @ 6.3.. I used live view, delay was approx 5 sec. I used wifi thru tablet to take photos.

It's a very good idea to take pictures without touching the camera.

However, there are some devils in the details. With your camera, shooting from live view probably means that the mirror slams down, the shutter closes, the mirror slams up again, and then the shutter opens again to start the actual exposure. The result of all this mirror and shutter motion is that the camera and lens bounce around for a while on top of your rail, like the head on a bobblehead toy. This explains why your images show something like 8-12 pixels of movement blur.

In previous communication, I explained to you as follows:

However, when you start getting images, you'll discover that at 10X, vibration is a huge problem. That's because it takes only 0.001 mm of movement at the end of the objective, to produce a 2-pixel smear back at the sensor. Your camera does not provide electronic shutter, so you will inevitably be having to deal with mechanical shutter vibration that will blur your images. For most people, the best solution for that problem is to use electronic flash, dialed back to low power so as to get short light pulses. A typical consumer-grade flash, dialed back to 1/16 power, will produce light flashes that are around 1/5000 second, and still bright enough to give a good exposure at base ISO setting. Diffuse that with some paper or a foam cup or white plastic yogurt container placed close to or around the subject as suggested by Deanimator, and the illumination will be in good shape.

What do you have available, for electronic flash?

--Rik

hotrodder19 · Post by **hotrodder19** » Fri Jul 12, 2019 6:09 am

Rik, I have a few flash units but including an SB-700 nikon unit. Are you suggesting flash instead of the Jansjo lights ? For what reason. What other lenses other than the F4 200mm lens could I use to get the whole insect in view ? I have 1oomm 2.8 macro lens. 50mm lens, 2x Kenko enlarger and other tele lenses Any suggestions ?

Cheers hotrodder19