The Microscopic Beauty of Plants and Trees

[Robert Berdan]

I am attaching some images of Plants and Trees. I started sectioning plants using various types of razor blades and hand microtomes and I am able to cut 30 micron thick sections which are Ok if I use focus stacking. But I am looking to purchase an affordable sliding microtome in the future to make much thinner sections like those in prepared microscope slides.

I posted an article on my site that shows some of the tools, techniques I have tried along with many original papers downloadable as PDFs for those that might be interested. Some of the stains like Safranin and Astra blue I found are not easy to get or have shipped to my home residence.

I used a variety of techniques, Polarixing, DIC, Darkfield, Fluorescence and Bright field mictoscopy and several different microscopes. The plants were from my fridge or yard.

Article for those interested: https://www.canadiannaturephotographer. ... ants1.html



Spruce branch hand cut and focused stacked - Toluidine blue stain


Pine wood section, polarized light with full wave retarder, from prepared slide


Cannabis sativa male leaf section by hand using razor blade - Darkfield microscopy, Cannabis is now legal in Canada


Potato starch grains in polarized light microscopy




Wide variety of blades I am testing along with a variety of hand microtomes to cut thin sections of plants. Many woody stems I have to soften by boiling in 10% glyercol until the wood sinks - I am having fun learning more botany and various techniques. This summer I will section and stain almost every plant I can get my hands on.

[Admin edit 3/1/2020, RJL: formatting and copy caption info from email]

[abpho]

Amazing. Thanks for sharing. You have some great images I that article. I'll have to revisit it when I'm not on m my cellphone.

[Sumguy01]

Very nice.
Thanks for sharing.

[Robert Berdan]

Today I tried cutting woody stems from a Popular tree using an old hand microtome from the 1950s using a disposable razor microtome blade attached to microtome blade I purchased on Ebay from India and I was able to cut 10-15 micron thick sections and was quite excited. I might not have to purchase an expensive sliding microtome after all - I will keep practicing and now I am going to experiment with some other stains like Astra blue and Safranin.

[iconoclastica]

Beautiful!

[Lou Jost]

These are extremely beautiful, and even the high-m ones look very sharp. It makes me curious about the optics you used. I did not see any mention of that in your linked article, (which was otherwise very informative!)

[ChrisR]

Somehow I missed these. This is excellent.

DO look at Robert's article:
https://www.canadiannaturephotographer. ... ants1.html

[ChrisR]

@R.B
I have a Reichert-Jung sledge microtome which I'm not sure I'll ever use. Its blades are robust angled things, which would be time-consuming to keep resharpening. Do you know if any of the replaceable blade types are suitable in general for sledges?

[Robert Berdan]

I use several microscopes. My 45 year old Olympus E which I purchased about 45 years ago and a more recent purchase - Zeiss Axioscope which I almost had to get a mortgage on :-) I photograph with Nikon DSLR cameras using free software Digicam control available on the Net for PCs

The small AO hand microtome cuts nice sections about 20-30 microns thick by hand. I recently purchased a sliding microtome from Radical in India for $500 US but haven't had a chance to test it yet. I received several other quotes for sliding microtomes, starting at $3000 up to $12000 which were just to expensive for me. Even used sliding microtomes on Ebay started at about $3000 plus hundreds of dollars for shipping because of their weight.

I plan to section more plants in the near future.

[Lou Jost]

Thnaks for the info about the techniques. I was more curious about the objectives though; what are your favorite ones for this work?

[Robert Berdan]

Hi Lou - The objectives on my Axioscope are infinity plan objectives, but I use a wide variety of objectives mostly Olympus 160 mm and have had success with all of them. Plan Acrhomats are my favourite because they offer a flat field, I have one Apochromat and several Fluorite. My Olympus Phase objectives are ordinary achromats - though I try to use the center of the image with these.

I use Photoshop and believe its can make any image look better and RAW files offer more flexibility.

Cheers
RB

[Lou Jost]

Thanks Robert for that info; yes, Photoshop + RAW can work wonders!

[Robert Berdan]

Hi Chris in my picture above I show a disposable blade holder, available from India via Ebay for $150 it fits regular microtome blade holders and you place the disposable blades inside it - I love it. Cuts really nice, does not require sharpening. The blades are $100 for 50 but worth it I think. I will try the cheaper cutter blades in the future as several researchers told me they get good results with them - blades are availble on Amazon.com

Cheers
RB

[ChrisR]

I've been told Japanese Feather Brand shaving blades are noticeably better than the rest. Whether the "proper " disposable microtome blades are any better than those, I have no idea. This holder holds single or double edged blades or utility knife blades. Do you think it might work for a slider? It sat unidentified on eBay US for months.

[Robert Berdan]

Hi Chris the feather disposable blades are very sharp and when I put them in the disposable blade holder from Radical in India - I got my thinnest sections yet. I am planning on using it on my sliding microtome I purchased from India as I don't want to spend a lot of time sharpening blades - I would rather be sectioning tissues. I want to test the cutter blades as I see several labs that cut wood use them in their microtomes and their sections look very good.

No matter what blades you decide to use, I find that I am getting better the more I practice. My goal is to cut 7-12 micron sections consistently. After that is choice of stains and little things like using magnets on the coverslip to flatten the sections when making permanent slides that seems to make small but significant differences. I am still learning as I am rather new at this.
RB