My Macro Photography Rig

I'd like to get into Microscopy.  Specifically, 10X to 20X magnification (but up to 100X) of abstract subjects, printed large on fine art paper and canvas.
  1. Sample images
  2. System
Note, you can click on any image to enlarge it.
 
 Sample Images

Butterfly Wings
Armenia, 12 Apr 2019
Here is a closeup of the wing of a Sunset Moth, taken using my Mitutoyo 10X objective.  Note, this is just a quick-and-dirty image.  I don't think I have found the best tube lens yet (I'm still testing my options).  And I need to work on lighting.
 
I could see making prints of mosaics or collages of say, these scales from half a dozen or a dozen different subjects.  Printed large in a grid on canvas.  Or eyes (human, mammal, insect) arranged in a
4x3 grid - printed large, sort of looking back at the viewer.  A study in the evolution of eyes, for example.

Crystals
Armenia, 6 Apr 2019
Crystals can be grown on a microscope slide and then photographed using polarizing filters.  There is an effect called Birefringence that results in amazing patterns in some materials.
 
Lake Placid, 2 Jun 2019
Below are some images I have made.  They are from microscope slides prepared by yodalovedme (eBay).  I will prepare my own slides in future.
 
The images at 20X magnification are approx. 2 mm across.  The ones at 10X are approx. 4 mm across.  Please click on an image to enlarge it.  The first one on the left is presently my favorite.
Hippuric Acid
20x magnification
0021
20x magnification
0022
20x magnification
0023
20x magnification
0024
20x magnification
0039
20x magnification
0046
20x magnification
0041
20x magnification
0042
20x magnification
0047
20x magnification
0048
20x magnification
0040
20x magnification
0043
20x magnification
0044
20x magnification
0045
20x magnification
0046
Scroll right >
Ascorbic acid (Vitamin C)
20x magnification
0001
20x magnification
0002
20x magnification
0003
10x magnification
0004
Saccharine
10x magnification
0005
10x magnification
0006
20x magnification
0007
20x magnification
0008
GABA (gamma-Aminobutyric acid, an amino acid)
10x magnification
0009
20x magnification
0010
20x magnification
0011
20x magnification
0012
DHEA (Dehydroepiandrosterone)
20x magnification
0013
20x magnification
0014
20x magnification
0015
20x magnification
0016
Potassium Chlorate
20x magnification
0017
20x magnification
0018
20x magnification
0019
20x magnification
0020
Aspirin
20x magnification
0025
20x magnification
0026
20x magnification
0027
20x magnification
0028
Pregnenolone
20x magnification
0029
20x magnification
0030
20x magnification
0031
20x magnification
0032
Caffeine
20x magnification
0033
20x magnification
0034
20x magnification
0035
20x magnification
0036
Resorcinol
20x magnification
0037
10x magnification
0038
Below is a sampling from Robert Berdan, Loes Modderman and others.  None of the following are mine.
Paracetamol and Saccharine in water solution, pol+ret
some amino acids from a fitness supplier
TRIS, water solution, pol+ret
Benzoic acid, Salicylspiritus and Tartaric acid
callus remover in aceton. 10x objective
callus remover in aceton, pol+ret
Urea, Glycine and one other, water solution, 10x objective
Magn sulphate, water solution
Niacinamide (Vit B3) and L-Asparagine, water solution. Pol+ret, 10x objective
Vit C and Na carbonate
Citric Acid
Src
Urea
Src
Urea,coffee?,water
Src
Niacinamide (Vit B3)
Hydroquinone,alcohol
Src
Resorcinol
Src
Ascorbic acid,water
Src
Tartaric acid
Aspirin
Resorcinol, alcohol
Ureum, water
Urea, water
 (Lots) More ...
Magnesium sulphate
Paracetamol
Hydroquinone+an addition
Tartaric acid and Saccharine
Urea, Paracetamol
Src
Aspirin and Paracetamol under polarized and oblique lighting
Src
Tartaric acid, Dextrin
Na tetraboraat and Glycine
Paracetamol
Paracetamol
Paracetamol
Paracetamol, betamethason
Paracetamol, betamethason
Aqueous resorcinol
Src
Betamethason, paracetamol
Src
Resorcinol
Urea + an acid
Paracetamol + addition, alcohol
Urea + addition, water
N/A
Betamethason/paracetamol A
Barnsteenzuur
Copper Sulfate
Epsom salt
Src
Epsom salt
Epsom salt
Urea
Copper sulphate, in gel
Ammonium di hydrogenfosphate
NAS
Tartaric acid
Salicillic acid and Propylhydroxybenzoate from alcohol solution
Urea and citric acid
Urea + citric acid, water solution
Na thiosulfate, water solution
Urea
Glycine
Progesteron
Tartaric acid after some time
Propylhydroxybenzoate and paracetamol
Thiocarbamide and three others, water solution
Malic acid and softener
Boric acid, mainly
Hydroquinone
Paracetamol, Alcohol solution
Progesteron
Resorcinol M
Hydroquinone and 2 other components
Resorcinol A
Propylhydroxybenzoate
Imidiazole
Malic Acid and Ibuprofen
Atropine
Penylephrini hydr
Efedzine / Hydroquinone
Tartaric acid and menthol
Hydroquinone and Urea
Resorcinol
Paracetamol
Paracetamol and wodka
Boric acid, Sacchanrie and at least 2 others, from water solution
4-hydrobenzoic B 2
TRIS and Glaubersalt, mainly
Oxalic acid
Tartaric acid, in water
kaliumwaterstofphatalaat A
kaliumwaterstofphatalaat
TRIS
Thiocarbamide and Rochellesalt
Mainly Acetylsalicylic acid
Resorcinol and Oxalic acid
Thiocarbamide and an industrial salt
Epsom
Recorcinol, alcohol solution
Resorcinol
"Snus is a traditional form of smokeless tobacco in Sweden. Images are an airdried water extract of a typical bergamot-flavored snus (pouch soaked in water for a few minutes). I am guessing the crystals are mainly salt. Polarized light (no retarder). Image is a stitch of 16 images. 4X objective."
"BIG crystal growth (if that's what it is on 1mm²) from Hydroquinone and a soft drink. Fascinating because they never touch other groups of similar crystals, but intrusions of a different kind are added without a problem. For people who want to learn something about crystallization !"
"The proportion of the two solutes is critical to achieving these forms - I generally start from a solution of 4:1 (v/v) of beta-alanine to l-glutamine and vary concentrations until I get something that works. Ensure that your slides are extremely clean - if they are even slightly hydrophobic the results are generally unusable."
Urea and an addition, water solution, 150x
Thiocarbamide and Rochellesalt
Hydroquinone, alcohol solution
p-hydroxyphenethyl alcohol
4-hydrobenzoic acid G
8-CHINLINOLSULFAAT
8-CHINLINOLSULFAAT
Omeprazole
3/4 hyldroxy phenyl G
Urea, Gluconic acid and an addition
Ijzermagnesiumnitraat
Resorcinol and vitamin drink
Boric acid, Sacchanrie and at least 2 others, water solution
From a garden fertilizer
Diclofenac, water solution
TRIS in water solution
Oxalic acid with resorcinol
TRIS, water solution
Resorcinol. water solution
Pyridoxin and Betaine
3/4 hyldroxy phenyl
Resorcinol, alcohol solution
Oxalic acid and an energy drink
Coffeine and Saccharine, water solution
Urea and gluconic acid
Saccharine and Dextrin, in water solution
Succinic acid and Gluconic acid, water solution
Testosteron Propinate C
Acetamide
Urea and others, water solution
Tartaric acid
From several acids
Urea
Gallic acid and Hippuric acid, water solution
A callus remover
Saccharine and Dextrin, in water solution
Resorcinol, thiocarbamide and a drink, water solution
Hydrochinon in stripes, after a drop of drink, alcohol solution
4-aminobenzoëacid
Nicotinezuur A
Na hydroxide in a special mixture, water solution
Guanidine HCL
n/a
Citric and Ascorbic Acid
Citric and Ascorbic Acid
Thiocarbamide and another component
A sheet of gelatin, stereo, 20x, multicolor filter
DL Tropiczuur A
From several acids
"Extract of white snus. White snus is based on the traditional Swedish oral tobacco product but instead of containing tobacco, it contains a gum base to which nicotine and flavours have been added (mint flavour in this case). I took one sachet of snus and allowed it to steep in a 50:50 mix of water and alcohol for 10 minutes I then filtered the liquid through kitchen towel before leaving to airdry on a slide for a week. Images are taken using x4 objective and simple polarized light (no retarder). Slide shown as well for reference."
Malic Acid + sugar
Tartaric acid, with and without retarder
3/4 hyldroxy phenyl
Benzotriazole
3/4 hyldroxy phenyl
Urea and Boric acid
Nicotinamide
Paracetamol and Salicylic acid
A "special concoction", water solution
Hydroquinon and other components, Alcohol
Resorcinol
Hydroquinone with an addition to straighten them out, alcohol and water
Hydroquinone, mostly
Saccharine, water solution
Nicotinamide
Nicotinamide A
Tartaric acid
Fertilizer, water solution
Malic Acid + sugar
Malic acid, from water solution
KA-Chloraat
"Copper sulphate, taken 5 seconds after each other. First the small crystals appear, and then a wave takes over in a second. In the meantime the crystals keep growing, as you can see in the right corner. This is typical for this chemical."
Magnesiumchloride
4-hydrobenzoic acid
Sulpher
Sulfur
Hydrochinon and glycine, alcohol solution
Progesteron, water solution
magnesium sulphate and several others
Sweeteners and Glycine
Magn sulphate, water solution
Paracetamol/vanille B
Vanilla b
Resorcinol AK
Hydroxy B
Hydroquinone and Glycine
Vanille/paracetamol D
Vitamine C/Salicylzuur
Vanille/paracetamol
Magnesiumchloride
Creatine and Na thiosulphate
Hydroquinone D
Benzotriazole
Magnesium sulphate, water solution
Antron
Bemegride
Ammonium Sulphate
Gluconic acid, water solution
p-hydroxyphenethyl A
Boric acid and another acid
Crystals that would need stacking. Combination of three colorful chemicals in water solution, 150x
Hydroquinone and Thiocarbamide
Magnesium sulphate, water solution
Hydroquinone and Thiocarbamide
Resorcinol, alcohol, 150x
Na-methoxyde
Na-methoxyde
Na-methoxyde F
Na-methoxyde C
Urea and Boric acid, water solution, 150x pol+ret
Amm thiosulphate and Na thiosulphate, water solution
Resorcinol and Tartaric acid, alcohol solution, 150x
Boric acid and Thiocarbamide
Callus remover liquid
A mixture of sugars, water solution
Boric acid and Urea, millions of little crystals popping up. Water solution
Urea and Boric acid, water solution. Less them 1mm²
Urea and some acids
Thiocarbamide and Ascorbic acid, water solution, pol+ ret , 150x
Na-methoxyde C
D-threo-2-amino
Pyrocatechine
Boric acid and Thiocarbamide, water solution, 150x
Resorcinol and Tartaric acid, alcohol solution, 150x
D-threo-2-amino.B
Gluconic acid
methoxyphenyl
Several kinds of fertilizer, water solution
D-threo.amino-1 A
Several acids
Malonic
Hydroquinone, Glycine and wodka
Oxalic acid pattern, alcohol
One of 20 years ago, from Urea , water solution
Resorcinol, alcohol solution, pol+ret, 100x
 System

My current system (a work in progress)
Lake Placid, 7 Jun 2019
Here is my current system.  Left to right: a new PC and monitor (for post processing), the old PC and monitor (for capture), the camera.

Regarding the camera (left to right):
  • 3D-printed holders for the polarizing, retardation and Rheinberg/darkfield filters
  • A 3D-printed holder for an LED light source
  • The X-Y stage through light source are mounted on an Arca-Swiss rail to a pair of Stackshot rails arranged in an "X" to support stitching (panoramas)
  • Stackshot Z rail (forward and back for focus stacking)
  • A Thorlabs optical breadboard, resting on Sorbothane feet on a fairly solid IKEA dresser on thick carpet on a concrete slab

The X-Y rotating stage makes it easy to move the sample up & down and left & right.  Then to rotate the subject to pose it.
 
The optical breadboard is isolated from my tabletop on the left on which my monitors and keyboards rest.  Still, I leave the room when I'm taking a series of photos (which can be a couple hundred) of say a butterfly wing for focus stacking.
 
At the 42MP resolution of the a7RIII (7952 x 5304px), they should print at:
  • 400 dpi at 13" x 20" (utilizing the full width of my 24" canvas for the width)
  • 300 dpi at 18" x 27"
  • 265 dpi at 20" x 30" (utilizing the full width of my 24" canvas for the height)

I will update this section periodically as I develop the system.
References:
   - 52MM STUDIO SETUP (after which I modelled my setup)
   - Matching Camera to Microscope Resolution
which I've plotted here -->
The dotted line (at Pixel Size = 4.5 µm) corresponds to my a7RIII (not counting the presumed benefits of Pixel Shift).  My Mitutoyo objectives are marked (most of those points are interpolated).
 
The Mitutoyos have a decreasing cost-performance benefit to resolution at higher magnifications.  It is possible that the benefits of these objectives at the higher magnifications are increased Working Distance and reduced Chromatic Aberation (due to them being APO).
 
I think this plot shows that my a7RIII is capable of
out-resolving my Mitutoyo objectives (but not by much for the 10X.. not counting the benefit of Pixel Shift). 
 
Note, this plot is based on traditional microscope design (the text refers to aligning the condenser and matching the condenser's NA to the objective's NA), so I may be mixing apples and oranges.
My Target System
Armenia, 25 Apr 2019
Goals:
  1. I'd like to use this as a learning exercise.
  2. I hope that using high quality components will let me cook up a system capable of high quality images.
  3. I want to be able to make large prints.  Thus, "stack & stitch" using the Stackshot rails for X, Y and Z.
  4. I hope to find and adapt older, slightly used, high quality components from eBay, etc.
  5. Speed up my whole image creation process.
Optically, this is the sort of system (left) I'd like to initally emulate using my components (right).  That is, add to my current system the linear polarizers, retardation
material, a light source and whatever bits of 3D printed holders I need to glue it all together (the grey and yellow parts in the animation).

I feel it takes a long time to process a large stack - converting the Raw Pixel Shift images from the camera to TIFFs for Zerene being much of it.  It requires several manual steps - easy drag-and-drop operations, but I have to be there.  I'd like to speed up the process by souping up my computer system and removing the manual steps.
 
Computationally, this is the system I'd like to build.  My goal is to automate the end-to-end process.  And convert and stack the images as they come off the camera.  To accomplish that, I am putting together a Multi-processor (MP) from single board computers (Raspberry Pi's)
and planning to build a fast PC (PC2) to do the stacking and post processing.
 
I'm planning to locate the MP, NAS and Ethernet switch in a separate room to avoid their cooling fans from affecting the image capture.  Everything is connected together by Gigabit Ethernet.  There is also a Wifi connection from the Controller on PC1 to a web server-based monitor so I can check on status using my laptop, tablet or phone.
 
Status: My new MP monitor based on my old Raspi Monitor is coming along.
 
References:
   - Old Microscopes
            THE BEST "AFFORDABLE" PETROGRAPHIC MICROSCOPES OF THE 1970s TO 1990s
Leitz Dialux, Early 1970s many parts and accessories are available on the market
American Optical Series 110, Mid-1980s [a so-so review]
Zeiss Standard, Late 1970s higher level are the Universal or Photomic models
Nikon Labophot, Mid-1980s an "entry level" benchtop scope, less expensive than the Optiphot line (which are rare in POL), unsurpassed optics, large controls
Wild Heerbrugg M21, Early 1970s considered by some experts to be the best microscope ever made
Olympus BH-2 BHTP, Late 1980s "the microscope I prefer over all others"
            Olympus BH2
   - New Microscopes
            Olympus CX31-P
   - DIY Microscopes
        Bertrand Filters
            New Bertrand Lens on Ebay
        Condensers
            Substage Condensers
        Lighting
            Tungsten halogen vs white LED
            Optiphot modular light source
            FAQ:How (and why) to use electronic flash at the microscope?
            Microscope Conversion to LED-Light
            HOW BRIGHT IS YOUR LIGHT PART III: LED CONVERSION
        Rheinberg Filters
            Making Rheinberg illumination discs
            How to Make Rheinberg Filters
            Rheinberg Filters for Photomicrography
   - Technique
            A Practical Guide to Creating Superresolution Photos with Photoshop
Raspberry Pi MP
Armenia, 15 Apr 2019                 Implementation
I'd like to build this Multiprocessor out of the latest version of the Raspberry Pi (Raspberry Pi 3 Model B+).  The Pi 3 B+ is clocked at 1.4 GHz, has 1 TB of RAM and supports Gigabit Ethernet at 300 Mbps.
 
I will model mine after Joshua Kiepert's RPiCluster.  He has documented his system well, including Eagle files for his power distribution cards.
 
At the end of that article, it says his 32-node system (using an older version of Raspberry Pi) does 10.13 GFlops, where "the first Cray-2 supercomputer in 1985
did 1.9 GFlops".  I look forward to benchmarking mine.  Let's see, Summit did 200,795 TFlops in the latest Nov 2018 Top500 List.  How close will I be?
Ethernet Switch
Armenia, 12 Apr 2019
I'd like to be able to support up to 48 Raspberry Pi's.  Add a port for the Master Node, two for the NAS, one for the PC1+PC2 (that I'll bring in from a separate switch where they're located at
my desk).  Making 52 ports all together.  This 52 port switch looks good.  Gigabit speed at a reasonable price.

Surge Protection
Lake Placid, 1 May 2019
I read that Ethernet Switches are vulnerable to power glitches.  I see many refurbished switches for sale.  There seems to be a fair amount of thunder storms in my area.  I had a bunch of electronics zapped last year in a thunder storm.
 
I assumed that meant I should get a battery backup, ie. an Uninteruptable Power Supply (UPS).  So, I estimated the power requirement of a UPS as follows:
  1. 46W max for the switch.
  2. 5.7W x 48 = 274W for the Raspberry Pi's.  (using the max. of 5.7W, 5.1W and 0.980 x 5.1V = 5.0W (400% busy, no attached devices)).
  3. 40W for the NAS (source).  Although that doesn't include the optional DX517 expansion unit.
The total of 360W+ lead me to this 450W CyberPower CP800AVR (which was highly rated in this review).
 
Great, but that review also said,
"Unfortunately, most affordable UPS units don’t offer much protection compared with a dedicated surge protector. In previous tests, electrical engineer Lee Johnson took apart our UPS samples to examine their guts. Based on his assessment, we found that our picks should protect your equipment about as well, if not for as long, as basic surge protectors we’ve tested before."
 
So, here is their most recent review of surge protectors.  It has a pretty glowing review of the Furman Powerstation 8.  Pricey but hopefully worth it.
 
I've ordered 4 - 1 for this, 3 for my other electronics.

Network Attached Server (NAS)
Armenia, 25 Apr 2019
This one looks good.  Five bays plus an optional 5-bay add-on.  140 TB (70 TB + 14 TB drive x 5) capacity.  Two Ethernet ports.  Good reviews.  At a reasonable price.
A fast PC (PC2)
Armenia, 5 Apr 2019
This will run Zerene doing the image stacking (using Zerene's "capability to ingest and stack new images on the fly as they are acquired by a tethered camera").  PC2 also runs Photoshop and Zerene for post processing. 
 
I expect to use an Intel i9-9900K (8 cores, 5 GHz), 64 GB RAM, 1 TB SSD, Windows 10, a good calibrated 4K monitor.
Polarizer
Armenia, 7 Apr 2019                 Tests
I want to make polarized photos like the crystals above.  It looks like that calls for 2 polarizing filters, a "retardation or wave plate" (mica, scotch tape or I'm going to try this retarder film) and a holder for the microscope slide. 
 
I'm planning to 3D print a holder for them.  Click on the animation ->
One polarizer (called the Analyzer) and the Retardation Plate would be located in the tube between the tube lens and the objective.  The Slide and other Polarizer
Glass polarizers
Retarder film
A 3D-printed holder
with condenser
would be held in the 3D printed holder.  The panel in this animation is an LED light source such as this one.
FIN