affinity photo Focus Stacking / Microscopy

[Graham L Furlonger]

Hello Forum,

I am posting this because I searched on "Microscopy" "Entomology" and "Focus Stacking" and saw few if any results, I may be in a minority?......

 

Having spent many years perfecting Microscopy with Entomology and recently converting from a well know rival, I was impressed with the power of Focus stacking within Photo. These two images were taken with modest equipment and a Canon 50d digital SLR dating from 2008 and a Brunel MX1 Stereo Microscope with 1x Objective, 10x Oculars, Giving resolution power of 10x, plus magnification of the camera. Whilst I have DSLR's with much higher pixel resolution, it is often not required and extends processing time, the 15 mp sensor 

These two images comprise multiple shots (35 & 24) focus stacked within Affinity Photo.

I won't bore you with the technical details, but if you want them you can message (I am happy to share)

Affinity has saved me exporting my images to a third party software which has enhanced speed of operation and ease of use, one thing to note is that the Affinity software is not too picky on the order of focus slices presented, which is a major benefit to other rivals.

Of course this technique could be useful to many area's of photography including, jewellery for record purposes, medical, veterinarian, instruction manuals and service documents where close up work results in very narrow depth of field.

 

Enjoy your photography and enjoy using Photo.

Regards

 

Graham.

 

 

[Patrick Connor]

@Graham L Furlonger

Welcome to the Serif Affinity forums

Those are wonderful. using tagging I found these other examples for Focus Merging of focus stacking but there are not many so this is a treat.

[John Rostron]

Very impressive!

John

[Reinhard Grossmann]

@Graham L Furlonger

Very impressive. I've just started to use Affinity Photo, and your example motivates me to compare eventually focus stacking by Affinity with Canon DPP. 

[unni]

@Graham L Furlonger

Very good work.

I have done some work in this area of focus stacking with homebrew setup. The setup details are in the post below.
For an year or two, I have not done any serious stacking work because it needs lot of time and energy to setup the equipments
and then put it back to place after the work. It takes about 4 hours for me to complete the photography part and then
another 3 hours to complete processing and editing. AP does a fantastic job in this field of stacking - performance,speed and ease.

Best regards,
Unni

 

[Graham L Furlonger]

So, I have been at it again! 

Ferdinandea Cuprea, Stereo Microscope 20x Magnification. 36 Individual images merged in focus stacking software. (Affinity Photo)

i was curious to see how finicky AF was to sequential order, from my own experiments it doesn't seem to matter.  

Without boring you all, here is a synopsis of getting the cleanest images with AF.

• you must lock down equipment so no unwanted vibration or movement between frames.
• ensure the camera (I use both Nikon FX D600 and Canon 50D DSLR ) is set to manual exposure.
• I perform manual white balance but auto white balance seems to be ok.
• I always shoot RAW and process in 16bit TIFF as lossless. I then convert to low res JPG for posting on social media etc. 
• ensure a good border when starting around your specimen to allow for any vignetting or post process errors. This is where using a larger pixel content than required gives the ability to crop leaving a good sized image.
• ensure that no natural light effects the exposure, I took a set of images once starting by sitting and then got bored so decided to stand up. The standing up images were one stop over exposed due to the light I was shielding by sitting down! 
• Ensure that you know your kit well, I always start with the stage highest IE. Focusing on the furthest part of the image and focus through the image lowering the stage away from the optics. This ensures no accidents with your specimen kissing your objectives!
• You can rescue some imperfections with post processing but it is very difficult to improve focus sharpness, don't be tempted to over sharpen using unsharp mask etc as this will destroy your tonal range and look artificial.
• I have tried lots of focus stacking software, even those for astronomy but I am finding Affinity Photo is coping well for my needs.
• I leave all my Microscopes (3) permanently set up. Why? Because I know from experience I am more likely to produce work if the setup and completion time is quick and hassle free.
• I personally do not use computer tethered software only because when MAC OS went to Catalina the Canon software I was using with my 50D stopped working, I wrote to Canon and received a reply saying they would look into this but alas still no result. I am currently transferring images to my Imac by Memory card and then transferring all the RAW images to hard drive prior to processing as the read time is much faster than directly from Mem card.
• Enjoy this hobby, its been a lifesaver during this time of lockdown in the UK.

regards Graham.

[Graham L Furlonger]

A few more images, hot off the press!

Comments very welcome, construction criticism only please. Thank you.

 

Details:-

Microscope:- Leitz Laborlux K, Optics (Achromatic Leitz 4x, 10x, 40x, 100x Oil) plus Bresser Achromat 60x 

Shot using the 10x Eyepiece with Brunel Microscopes Adapter and either,

Canon 50D APS C or Nikon D600 FX full frame.

All images imported into Affinity as RAW files.

Processed in Affinity Photo using Focus Stacking Method.

Borders and text added in Affinity post processing.

Note:- the Darkfield used for these images is a DIY home made patch stop mounted onto 1mm the Clear Optical Polycarbonate suspended under the condenser stage with a simple clothes peg! (you need some originality with these things!)