Beginner questions: Part 11: 3rd session with Bob's printed knife holder

[deBult]

Adjusted Photo set-up: stitched 9 pano slides into 1 overview.

(Note: pano stitch as workaround to build an overview picture as I'm using a 4* objective plus micro 4/3 camera plus 2.5* NFK projection eyepiece)

SHK and WAsimIII: slightly thicker cut.
Next step: 50 cuts with SHK and 50 with Bob's printed holder (to practice cutting and compare).

Bob, Jorg, Klaus: thanks for tips and guidance: you guy's are great!

Best,
Maarten

[Klaus Herrmann]

Hallo Maarten,

"not too bad!"

Unser großer Lehrer Dieter Krauter hat vor vielen Jahren einmal gesagt: "Üben sie, Üben sie  wenn sie mal 1000 Schnitte gemacht haben werden sie sehen: es geht schon!"

Schöner Schnitt und gelungene Färbung!

[Fahrenheit]

Guten Morgen Maarten,

da kann ich mich Klaus nur anschließen und danke für die Blumen.

Herzliche Grüße
Jörg

[deBult]

Next step: feed back appreciated!

As per Klaus and Jorg's challenge 50 cuts with Bobs printed knife holder + OLFA blades and 50 with SHK + Leitz blades both using the "Scheidehilfe".

Hedera helix leave stem (petioles) this time.

Observations:
- Both series of cuts are slightly to thick
- New knives used after 25 cuts
- Need to change the camera setting for exposure and white balance to full manual next time
- I expect stacking will improve the picture
- All cuts with Bob's knifeholder went smooth (only 1 failure)
- Something must be wrong with the way I handle the SHK knife holder as 30 % of the cuts failed,
  and all of the cuts with the second knife required  100 % increase in thickness (picture is from the first 25 thin cuts)

Best Maarten 

[deBult]

A new run of cuts to see whether we can get thinner slides:
- 25 cuts: using Bobs printed OLFA knife holder, fresh knife, schneidehilfe, lots of alcohol during he cutting
- most cuts have a damaged centre
- Wasim III using the Klaus instructions (no heating)
- reduced the clearing: 5 minute in aqua dest: to see whether we can hold more of the red colour: no succes

Next try:
- "in between" cuts on thickness
- heating during the last minute of the staining phase
- try and prevent the "dirt" getting into the slides
  (think the dirt is caused by lint's of paper-fibre, as I used paper towels to clean the Euparal dropper in between sessions: the Euparal may be contaminated already,
   a the Euparal is nearly gone I will have to ask Klaus for a fresh bottle anyway)

Best,
Maarten

[Bob]

Hi Maarten,
that look very well now - nice to see that you continue with the effort! To find the perfect section thickness for object and sectioning process you might change to a 5 sections - review under stereo microscope - next 5 sections routine.
Did you find chrystals in your stem sections? I once have wrecked a blade in an instant by probably cuttling into one.

Concerning the dirt: to me your images look very clean - is that after a lot of cleaning work in image editing? There will be a point of diminishing return: You spend a lot of effort and only save a tiny amount of editing time.

Bob

[deBult]

Bob,

Tnx for feedback: appreciated as I have little comparison material and still have to learn what to look for ...

Have not encountered any crystals yet: good to know this can happen.

Still lacking a stereo scope .....

Actually no cleaning in photo-post-processing yet .
There is some dust on the slides as they are fresh, Euparal only partly dried: so I cannot clean them yet 

[Bob]

In this case I wouldn't worry about the Euparal and your process, to me your images are very clean.
"Clean" can mean a lot of different things when buying slides - since you look for perfection I would clean them myself before use. In this case here you don't need any special adherence properties so wiping with "Wundbenzin" will do nicely.
Slides out of older boxes (decades) often have damaged surfaces with a milky look that doesn't clean up any more.

[deBult]

Bob,

Tnx again.

I'm using relative older slides (got several 1000 for free), put a full box of 50 of them in a plastic slidebox with holes cut into the box and then the dishwasher .....convenient and helps to remove the white "haze".
Net result is perfect pre-cleaned object-slides.

Using Zeiss coverglasses 1.5 with strict thickness tolerance: clean/un dust them with an old cotton towel.

Thanks for the "wundbenzine" cleaning recommendation.

My issue is the lint and small dirt partikels introduced When building up the final slide (object-slide + sample + Euparal + cover glass). I eliminated one source of lint:  fibers from the paper towel used to clean the Euparal-dropper.

Plus the slides collect some dust during their 48 hours stay on the hot-plate and When I take picture after 24 hours (no patience to wait any longer) I cannot un-dust them without disturbing the -only partially cured slide...

Best