Flame of the woods

[unni]

Camera: Nikon D3100
Lens: 5X microscope objective
Number of frames: 223
Time taken by computer to process focus merge in AP : 1 hr 20 mts
Length of the yellow anther: 3mm
The vertical member is the pistil.
Rose color are of the petals.
Name of flower: Ixora coccinea

A picture of the full flower is also shown to get relative feel of the magnification.

 

[John Rostron]

Very impressive, but I wonder if you have gone over the top in terms of frames. 223 seems rather a lot for what would be 3-5mm range of your subject.  Having said that, it could be that you have automatic focus-stacking hardware which makes 223 shots easy. Did you use automated focus-stacking hardware? What hardware did you use? Wht was the numerical aperture of the objective?

 Must say that I am impressed that Affinity Photo can cope with 223 images!

John

[unni]

I am enclosing the pic of objective and extender rings. The dof is around 25 or so microns. So the more number of frames. I have used a homebrew setup with milling machine table. It's very heavy, 8" square size. Movement is manual by turning the handle. Next time I set it up, shall take a snap and attach. AP seems to process one image after another,in sequence. So it may never get stuck based on number of images except taking more time. I have not attempted trying small batches and then stacking the output of each stack . This way editing will be efficient for each group but I have not come across such edit requirements till now.i think hairs, crisscross objects etc will require such edits.

 

[unni]

[John Rostron]

Given your depth of field and the depth of your subject, then 223 images in your stack seems reasonable. Given your set-up you must be congratulated on obtaining such an excellent result.

It is many years (since I retired) that I had access to such equipment, but I still maintain an interest in these matters. Nowadays I confine my macro work to a limit of 1:1.

Focus-stacking in batches should, in theory give similar results to one big stack. But if Affinity does the job in one fell swoop, then stick with it.

I take it that Flame of the Woods is the name of the flower.

John

[unni]

Thanks John. Yes, Flame of woods is one of the flower's many names.

I am enclosing some of the pictures. Quickly made a mock partial setup sothat I can share some data with you.

The milling machine vice is available in machine shops. Heavy due to its 8" square size. 12 Kilograms.
The dial, I made from kitchen utencil aluminium plate (10 inch dia). In India, aluminium disc type plates are used to cover vessels. Its available
in various sizes. I think I used Inkscape to make the print out of the dial which has 360 divisions. Made different sections of it
in two or three A4 papers and joined it. The bent forceps is my dial pointer ! I just keep it on the table as shown and rotate the disc
by counting the divisions. For the pitch of the vice, 1 division gives me 8.66 microns movement. The blue base stand is a school burette
stand. The accessories also are parts of the stand. I keep the flash on it. The specimen is kept on the small scissor bench vice.
I use Godox X1N remote on the camera which will trigger the flash. Earlier, I used a cable. If using cable, you may have to cut all the wires
in the cable except the center terminal wire , ground and shield. The camera will then not see the flash. This is required because there is no lens.
So flash will not trigger if the remainng TTL connections are connected to the flash. A cable release is used to triggercamera. Flash kept at 1/128. It may
go up to 1/32or so if you use too many diffusers. Setting up and dismantling is the most time consuming and strenuous part. Clicking-turning-clicking is easy !
A similar setup can be made with other type vices also but this has a solid V groove which prevents wooble, cross axial movement etc. The larger the diameter
of disc, the easier it is to move through divisions. In some shots with  10X objective, I have gone by half steps which means 4.33 microns per step.
My objective is a metallurgical long working distance objective. You can also use biological ones designed for 0.17  coverslips though I have not tried.
Elsewhere I have read that it does not make any distance. I use Fotidox extension tubes. Bought two sets, mix and matched to get the correct distance
to objective flange from camera flange. Camera flange to sensor distance is available in net. The objective 160 means distance 150mm from objective flange to sensor.
In the picture, I have aligned the extension ring markings sothat its easy to understand. The RMS adaptor is available in aliexpress.Objectives are also available.
I have used Plan achromat. Semi Plan type, I have not tried. Plan APO types are very expensive.
Here is one link http://www.photomacrography.net/forum/viewtopic.php?t=12147

I did not go for infinity objectives because of complications related to tube lens.

Hope this helps if you plan to make a simple setup with finite objectives. @John Rostron

[John Rostron]

Thanks for all that @unniThis looks like dedication. I would be interested in seeing some of your other images.

John

[unni]

Thanks @John Rostron

The image below is from 200 frames and taken with 10X objective, processed in AP. Its a very small bee. I am reprocessing the raw files from repository with AP. I have only a few. This work needs around half a day including processing. I have to freshly charge the flash and camera batteries, etc. So eventually ends up with lesser of this category ! I think this was taken at less than 5 micron movement per step. SO half division in my setup.

[John Rostron]

Again, impressive. I have just been reading a thread on the Cambridge in Colour forums discussing the pros and cons of rack--focussing and ring-focussing (aka barrel-focussing). The consensus seems to favour ring-focussing. However with the equipment and at at the scale you are using, I do not see any alternative to rack-focussing.

John

[Graham L Furlonger]

On 10/21/2018 at 6:55 PM, John Rostron said:

Again, impressive. I have just been reading a thread on the Cambridge in Colour forums discussing the pros and cons of rack--focussing and ring-focussing (aka barrel-focussing). The consensus seems to favour ring-focussing. However with the equipment and at at the scale you are using, I do not see any alternative to rack-focussing.

John

Interesting debate John, I have always been led to believe that barrel focusing caused more image distortion but I have no substantial proof.