Microscope Stylus Images
Background
Photomicrography and macro photography are both popular subjects in the InterWeb,
including among better writers than I.  Consequently, content here will focus (heh) on specific experiences,
rather than more general theory and technique,
which for macro photography see e.g.
Allan Walls.
Years ago, I saw and admired
Micrographia
and more recently
Ray Parkhurst's
work
including contributions to vinylengine.com
Stylus images on the not cheap (but not massively expensive).
Capturing images of phonograph needles combines 3 of my long-time interests:
audio, photography, and microscopy, although mostly microscopes with cameras.
While not aspiring to create aesthetically pleasing images,
facilitating capture of clear and useful phono stylus image wear images seems achievable.
Ray demonstrated that
Nikon's
long working distance CF objectives are well-suited to this,
while I already owned American Optical (AO) microscopes lacking long working distance objectives, oh well.
Generalities
Microscopes traditionally shared some mechanical conventions:
- RMS (Royal Microscope Society) Objective threads
- 23.2 o.d. eyepiece (ocular) tubes
- 160mm "tube length"
Consequently, optically-matched objectives and oculars could be swapped among frames.
Practically, after frames evolved from straight tubes to separate heads with prisms,
additional lenses might be introduced, specifically to allow for e.g. vertical illuminators
and/or polarizing filters to be inserted into optical paths.
These optics are typically optimized for some manufacturer's objectives optical formula.
Most modern microscopes use other than RMS threads;
here is a list.
Here is a list of threads used in astronomy.
Camera + Microscope Combinations
Brooke Clarke also addressed
these combinations.
Attaching microscope objectives directly to cameras is considered macro photography,
rather than photomicroscopy, and will mostly be addressed elsewhere.
However, attaching a camera body to a microscope body
with nothing but air between objective and sensor
is generally considered photomicroscopy.
This works with so-called finite objectives.
Infinity objectives want a so-called
tube lens to make rays finite,
either for oculars or directly focusing on a sensor,
in which case that tube lens may be a telescopic camera lens focused at infinity.
Since some infinity microscopes depend on their tube lens
to correct objective aberrations,
direct projection from microsope's tube lens
to camera's sensor may yield better images.
AO pioneered infinity microscope production in the U.S.;
read here for more history
Practically, for optical magnifications much more than 1x,
technique and patience exceeding mine are wanted for hand-held macrophotography.
For inanimate as well as opaque subjects, hand-holding has few advantages.
Opaque object microscopy requires reflective, rather than transmissive, illumination.
This is called episcopic (contrasted to diascopic) with illumination accomplished either by
external illumination sources or using so-called vertical illumination built into microscopes,
where light may be directed down thru objectives, e.g. using prisms or other partial mirrors
and called brightfield, or coaxially around objectives, called darkfield illumination.
Most conventional (diascopic biological) microscopes can be used with external illumination
for some opaque microscopy, but their objectives are optimized for viewing thru glass slide covers,
which becomes increasingly important for higher magnification objectives, e.g. more than 10-20x.
Live View, focus bracketing, Abbe
Using a digital camera with zoomed Live View eases focusing.
Beyond convenience, best viewfinder focus with DSLR mirror down may differ from best focus on the sensor.
Silent shutter (Canon's mode 2) and
remote shutter release minimize vibration during exposures.
Replacing halogen illumination with blue LEDs should improve image resolution, with
Raleigh's criterion having Abbe diffraction limit depend on illumination wavelength and
Numerical Aperture (N.A.).
Practically, blue LEDs not only have shorter wavelengths than do red and green
but excluding those longer wavelengths also usefully reduces chromatic artifacts from older and simpler lenses.
However, blue sensors are relatively sparse in digital cameras;  high resolution is required to compensate.
Canon DSLRs with Live View and silent shutter (e.g. 50D) can be had used for less than $100.
Even better,
free Magic Lantern firmware adds
focus stacking for many such cameras.
Eye tube adapters with typical T2 or M42 threads
require camera-specific adapters.
A projection ocular and extension tubes should cost less than $100;
monocular microscope heads typically cost much less than trinoculars,
while having less light loss than binocular heads,
and used eye tube camera adapter clamps go for around $15.
Good complete used microscope
with epi illumination and bright/darkfield objectives can be spendy.
Starting from scratch, consider
Nikon 210mm B/D objectives in an Optiphot 66;
many were used for semiconductor electronics wafer inspection...
baby steps with EPIStar
Initial stylus image capture employs a 10x epi microscope objective with darkfield illumination
and Olympus NFK 3.3x projection ocular into a Canon EOS sensor.
To hold the stylus at ~45 degrees, the cartridge was mounted in a headshell
with no finger lift and clamped at the bayonet connector using a clothespin:
Although it facilitates sorting spacing between camera sensor and objectives, a bellows is not necessary;
extension tubes suffice, with more length yielding greater magnification.
The resulting image has decent resolution, but poor contrast.
This microscope has short working distance objectives;
the 20x objective would not clear this stylus' plastic grip at this angle.
90 degree shots will clear but require image stacking;
whether objective darkfield illumination will then
highlight stylus wear spots remains to be seen.
EPIStar contrast
To investigate low contrast,
Ray Parkhurst suggested
removing the camera and looking down into the tube.
Internal surfaces were not reflecting appreciable light,
but a small bright spot and dim blob are seen in the optic at the bottom of the eye tube.
Lacking ground glass, wax paper was laid over the camera adapter:
The dim blob is revealed as the projected stylus.
The bright spot appears less focused than when viewed directly and,
depending on viewing angle,
more or less distinct from the projected stylus image.
Removing the microscope head and looking down into the arm
reveals a bright spot centered in an epi illuminator lens
in a port off to one side
as well as a less bright spot on the lower lip of that port.
That very bright area below the epi lens is the brightfield mirror.
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An A.O. #3002 image erector spaces the head further from the epi illuminator
and slightly improved contrast, despite adding magnifying optics in the path:
EPIStar 10X with EF-S 55-250mm STM
Infinity objectives can be used with a camera lens focused @ infinity,
in this case, a Reichert EPIStar 10x
with Canon EF-S 55-250mm STM zoomed to 250mm.
This is an example of camera telephoto as tube lens.
To prevent this zoom from collapsing to 55mm,
the camera body needs support.
Fortunately, focus is internal.
Some observations:
AO Series 10 - headless
Disappointed with EPIStar images, Series 10 configuration experiments resumed.
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RGB ring light
Microscopes better resolve using blue illumination, since shorter wavelength
and less opportunity for chromatic aberrations.
A 40mm RGB LED halo ring
fits around microscope objectives and can be supported by a 20mm i.d. o-ring.
That o-ring below LED halo blocks some illumination...
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Bayer-filtered camera sensors have relatively sparse blue pixels,
aliasing is liable for images with less than 4x oversampling.
Dark wedge along the center of the above blue M55E
stylus image is partly surface not in focus,
but also stylus angle not 45 degrees (twisted cantilever).  
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Glued a curled strip of business card
as a half-cylinder to the halo backside...
...an o-ring secures LEDs to the objective:
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Rotating the headshell in a clothespin brings
more of the stylus upper surface nearly in focus,
and using green light instead of blue employs more camera pixels.
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Contrast could be improved by putting something flat black between the cantilever and its light plastic grip.
calibration
AKA longitudinal chromatic aberration correction
Microscope fine focus is indexed in microns, 200 per revolution.
Images of 0.01mm test slide (with cover slip) using MEIJI S.PLAN M 20X
with Series 10 tube lens
and Olympus PE 2.5X; no matching corrections.
Numbers by colors are micon settings to focus that color:
red 144
| green 153
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white 148
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blue 153
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Red focus is out about 9 microns from blue and green..
This is axial (or longitudinal) chromatic aberration.
A.O. 10X with NFK 3.3X
With an AO head, AO objectives are substantially corrected, better matching a Nikon CF PL relay lens.
Headless, an NFK relay lens approximates AO tube lens corrections.
Finding the stylus with a microscope objective is easier using conventional oculars
than with camera Live View and relay lens.
Since the head has an infinity tube lens, this requires infinity objectives.
Since Series 10 AO Spencers often include a 10X objective, how well does one work on a stylus?
Because they are smaller in diameter than MEIJI and Leitz objectives
for which cardstock half-cylinders were epoxied to LED halos, a shim was cut from clear vinyl tubing.
This particular objective appears to have about 4mm working distance.
With halo illumination, a stylus tip is better illuminated with cartridge vertical centerline
rotated more nearly 60 than 45 degrees from horizontal, reducing surface in focus.
With NFK 3.3X relay lens and about 200mm projection distance,
full field of view with APS-C sensor is usefully in focus;
both of these images (downsampled, but not cropped) were captured after central focusing (zoomed Live View):
 
After visually confirming that minimal detail is lost by downsampling original (5472x3648) images by 3x,
then, given optics are NOT matched for color correction,
compared whether blue (left) or green (right) illumination yields more detail:
 
Approximate IrfanView color corrections settings for recovering dynamic range after grayscale conversion
and before 3x downsampling: brightness ~25, contrast ~99, gamma ~1.65