New to Me American Optical 815 Microtome

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rnabholz
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New to Me American Optical 815 Microtome

#1 Post by rnabholz » Sat Jan 02, 2016 10:51 pm

Hey all,

Thought you might be interested in a project that I have been working on.

I have been interested in acquiring a Microtome for about a year now. I have a hand held microtome, but something about the cool mechanism, machined parts, and cast iron really captured my interest. Very expensive new, I have been watching eBay for a deal on an old one. Most have sold around $200 or so, plus some significant charges for shipping as these weigh about 2.3 metric tons! Really heavy.

Recently an American Optical 815 unit came up for sale - the pictures in the listing showed that it was pretty dirty and showing some neglect. I expect that is why it did not sell in the first listing and what led the owner to relist at a very low price, saying that it was priced to sell. One click of the Buy it Now button and I had my Microtome!

Four days later, the worlds heaviest box arrived, and inside was my old/new microtome. Here is what I found.
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815Front.jpg
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815driveside.jpg
815driveside.jpg (127.61 KiB) Viewed 4883 times
815drivescrew.jpg
815drivescrew.jpg (118.32 KiB) Viewed 4883 times
815frontparts.jpg
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Re: New to Me American Optical 815 Microtome

#2 Post by rnabholz » Sat Jan 02, 2016 10:54 pm

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815knifeslot.jpg
815knifeslot.jpg (126.89 KiB) Viewed 4882 times
815specimenholder.jpg
815specimenholder.jpg (147.54 KiB) Viewed 4882 times
815knifeholder.jpg
815knifeholder.jpg (131.35 KiB) Viewed 4882 times
815knifecradle.jpg
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815crankside.jpg
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Re: New to Me American Optical 815 Microtome

#3 Post by rnabholz » Sat Jan 02, 2016 11:11 pm

The action was very stiff, partly due to being very cold as it had been riding around in the back of a delivery van in 20 degree weather, but also due to old and dried out lubrication.

There was paraffin everywhere, the entire machine was covered with a film of the stuff. So obviously a serious cleaning was in the cards.

I used Goo Gone for the largest part of the cleaning. It attacked the paraffin very well, as well as some of the dried lubes. To clean the rust, I used Evaporust- great stuff. Lots of nooks and crannies to get to, toothbrushes, dental picks, Q-tips all were employed at times.

I posted a message a couple of days ago about Pike Oil - that was the recommended lube according to the manual. No modern product bears that name, but I was able to find a reference that identified it as a Mineral Oil made as a lubricant and sharpening oil made by Norton Company. Norton is still in business and makes a mineral lube and sharpening oil. I ordered a can, and used it on all of the sliding surfaces.

Once the mechanism warmed up and was lubed properly, it ran like a swiss watch. I set the cut to 40 microns, cranked the handle 100 times and found the the specimen holder had moved 4 mm! Perfect!

Here she is all cleaned up
Attachments
C815front.jpg
C815front.jpg (119.12 KiB) Viewed 4881 times
C815Front2.jpg
C815Front2.jpg (118.08 KiB) Viewed 4881 times
C815driveside.jpg
C815driveside.jpg (241.81 KiB) Viewed 4881 times
C815back.jpg
C815back.jpg (113.49 KiB) Viewed 4881 times
Last edited by rnabholz on Sat Jan 02, 2016 11:40 pm, edited 1 time in total.

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Re: New to Me American Optical 815 Microtome

#4 Post by rnabholz » Sat Jan 02, 2016 11:14 pm

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Re: New to Me American Optical 815 Microtome

#5 Post by rnabholz » Sat Jan 02, 2016 11:22 pm

It was time for a trial run.

I placed the microtome knife that I had bought last year for use with my hand microtome in the knife holder.

I used a carrot as a specimen holder and celery leaf as the subject.

I won't tell you that I have it all figured out, but I was able to get a decent section and made this image.

I have a lot to learn, but it will be fun, and I have my very own Microtome.
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Celery Leaf 1-2-16.jpg
Celery Leaf 1-2-16.jpg (81.76 KiB) Viewed 4876 times
Last edited by rnabholz on Sat Jan 02, 2016 11:36 pm, edited 1 time in total.

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Re: New to Me American Optical 815 Microtome

#6 Post by Dale » Sat Jan 02, 2016 11:26 pm

What a fantastic job, and thanks for the exquisite photos.
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Re: New to Me American Optical 815 Microtome

#7 Post by Crater Eddie » Sat Jan 02, 2016 11:30 pm

Fantastic cleaning job! That beautiful machine looks like new! Well done sir!
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Re: New to Me American Optical 815 Microtome

#8 Post by JimT » Sun Jan 03, 2016 12:00 am

Congratulations. Looking forward to more images of sections. How thin can it cut?

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Re: New to Me American Optical 815 Microtome

#9 Post by KurtM » Sun Jan 03, 2016 12:23 am

Rod on the loose with that thing! I feel vaguely terrified and fearful, even way down here in Texas!! :shock:

Seriously, I very much look forward to seeing what Rod comes up with now that he has this baby in his arsenal. I've seen some pretty darn nice images come out of this guy, folks, using just whatever hand tools he had before he got his hand microtome. I think we may be in for some real treats.
Last edited by KurtM on Sun Jan 03, 2016 1:20 am, edited 1 time in total.
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Re: New to Me American Optical 815 Microtome

#10 Post by einman » Sun Jan 03, 2016 12:50 am

Very Nice. I purchased the same model sometime ago. There are pictures posted here on the forum. Fortunately I did not have to anything but oil it. Looking forward to sharing ideas.

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Re: New to Me American Optical 815 Microtome

#11 Post by rnabholz » Sun Jan 03, 2016 1:35 am

Thanks everyone.
JimT wrote:Congratulations. Looking forward to more images of sections. How thin can it cut?
Well, the setting scale goes all the way down below 10 microns, but I know that there is a lot more to cutting that thin than just setting the dial.

I found 40 microns using the carrot carrier a challenge. Need to learn a lot about the technique.

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Re: New to Me American Optical 815 Microtome

#12 Post by rnabholz » Sun Jan 03, 2016 1:42 am

KurtM wrote:Rod on the loose with that thing! I feel vaguely terrified and fearful, even way down here in Texas!! :shock:

Seriously, I very much look forward to seeing what Rod comes up with now that he has this baby in his arsenal. I've seen some pretty darn nice images come out of this guy, folks, using just whatever hand tools he had before he got his hand microtome. I think we may be in for some real treats.
Thanks Kurt, you are too kind.

This may be a bit more of a challenge to learn, but there are a lot of long winter nights ahead, so I will see what I can do.

Thanks again,

Rod

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Re: New to Me American Optical 815 Microtome

#13 Post by rnabholz » Sun Jan 03, 2016 1:44 am

einman wrote:Very Nice. I purchased the same model sometime ago. There are pictures posted here on the forum. Fortunately I did not have to anything but oil it. Looking forward to sharing ideas.
Great, I will dig up the thread.

Any pointers, tips, success stories?

Thanks,

Rod

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Re: New to Me American Optical 815 Microtome

#14 Post by zzffnn » Sun Jan 03, 2016 1:57 am

Great job!!
Selling my Canon FD 200mm F/2.8 lens

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Re: New to Me American Optical 815 Microtome

#15 Post by Charles » Sun Jan 03, 2016 2:33 am

Looking good Rod!

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Re: New to Me American Optical 815 Microtome

#16 Post by 75RR » Sun Jan 03, 2016 12:48 pm

Congratulations on the microtome.
Nice clean up. Perfect winter project.

Believe the sharpness of the knives is key to thin sections.
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Re: New to Me American Optical 815 Microtome

#17 Post by gekko » Sun Jan 03, 2016 1:22 pm

I just saw this thread. A very nice piece of precision machining and you've done an expert job of restoring it. I enjoyed looking at the result, and look forward for more. Thank you for sharing this sequence of pictures.

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Re: New to Me American Optical 815 Microtome

#18 Post by einman » Sun Jan 03, 2016 4:00 pm

rnabholz wrote:
einman wrote:Very Nice. I purchased the same model sometime ago. There are pictures posted here on the forum. Fortunately I did not have to anything but oil it. Looking forward to sharing ideas.
Great, I will dig up the thread.

Any pointers, tips, success stories?

Thanks,

Rod
No. I have spent some time looking for the best way to secure the samples. Cheaply! As there are several ways if you want to spend the money. I did read an article where you make a wooden block that mounts in the microtome. You then make paper molds attached to the block for the paraffin. After pouring the paraffin on/around the sample and letting it set you peel off the paper. Insert the block into the microtome and have at it. I purchased like 10 lbs of paraffin from an e-bay seller. It is sitting in a box. It was way more than I need but it cam as a single block. The quality of the paraffin also affects how thin you can slice the sample. Quality assessment is based in part on melting point.

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Re: New to Me American Optical 815 Microtome

#19 Post by rnabholz » Sun Jan 03, 2016 6:21 pm

Thanks zz, Charles, 75, and gekko


einman wrote:
rnabholz wrote:
einman wrote:Very Nice. I purchased the same model sometime ago. There are pictures posted here on the forum. Fortunately I did not have to anything but oil it. Looking forward to sharing ideas.
Great, I will dig up the thread.

Any pointers, tips, success stories?

Thanks,

Rod
No. I have spent some time looking for the best way to secure the samples. Cheaply! As there are several ways if you want to spend the money. I did read an article where you make a wooden block that mounts in the microtome. You then make paper molds attached to the block for the paraffin. After pouring the paraffin on/around the sample and letting it set you peel off the paper. Insert the block into the microtome and have at it. I purchased like 10 lbs of paraffin from an e-bay seller. It is sitting in a box. It was way more than I need but it cam as a single block. The quality of the paraffin also affects how thin you can slice the sample. Quality assessment is based in part on melting point.
Cheap is always good.

My interest is in prepping for short term observation, not that interested in permanent mounts.

With regard to melting point, is lower better?

I have used carrot as a carrier for plant matter, but, I think that you need fresh crisp carrots for good results. Of course, they are relatively cheap and widely available.

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Re: New to Me American Optical 815 Microtome

#20 Post by einman » Sun Jan 03, 2016 7:01 pm

Melting point should be about 56-59 degrees C. Quality varies. You can purchase lab grade but it costs about $14/kg. There is a grade referred to as Paraplast. They claim you can make sections down to 2 microns without the paraffin crumbling etc.

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Re: New to Me American Optical 815 Microtome

#21 Post by rnabholz » Sun Jan 03, 2016 8:47 pm

A couple more images from new sections of the celery leaf

The first is a single frame, the second is a 7 image stack
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Celery Leaf 1-2-16 II Single.jpg
Celery Leaf 1-2-16 II Single.jpg (94.67 KiB) Viewed 4802 times
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Celery Leaf 1-2-16 II Stack.jpg (81.53 KiB) Viewed 4802 times

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Re: New to Me American Optical 815 Microtome

#22 Post by mrsonchus » Sun Jan 03, 2016 11:17 pm

Hi Rod, that's a beautiful machine and a superb refurbishment by you! :D Congratulations! :D
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Re: New to Me American Optical 815 Microtome

#23 Post by rnabholz » Mon Jan 04, 2016 1:03 am

Thanks John, I hope to put it to good use.

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Re: New to Me American Optical 815 Microtome - UPDATED

#24 Post by rnabholz » Wed Jan 06, 2016 1:44 am

Well, I am claiming a small victory tonight.

A few nights fooling around with carrots and potatoes as a carrying medium and having not great success leads me to believe that they are just to flexible to cut this thin. 40 microns made baby food of both.

But that lack of success caused me to try a number of different remedies. Most notably I did sharpen my knife. Even after the sharpening mush was all I could get from the vegetable carriers.

So today I stopped by the grocery store to get some paraffin - I know, not exactly a scientific supply depot, but I hoped good enough for testing purposes. Good old Gulf Wax sold to most for canning, but not me.

I liberated a cube from one of the blocks and clamped it in the specimen holder. Set the cut to 40 microns and turned the wheel. The good news was that the knife is cutting, the bad news is that it was turning out little wax cigars. The wax was rolling into tight tubes.

I thought about it for a while, and it occured to me that it may be temperature related, as my little lair is a bit cool in the winter months despite my use of a small space heater. I grabbed the space heater and placed it on the bench and let a gentle current of warm air flow towards the Microtome, wax and knife.

After a few minutes I cranked the wheel and could immediately see the difference in the behavior of the wax off of the knife, with just a small prodding by me using a small paintbrush, a flat section unfurled. Another crank, another section and they were connected, crank, section, ribbon longer - THIS IS GOOD!

Cranked the cut thickness dial to 10 microns. Cranked the wheel and a delicate little section flowed off of the knife, kept cranking and the ribbon grew. The ribbon was not perfect, it showed some waviness, but I don't think it is variation in thickness, just a bit of "bunching" as it came off of the knife. If it indicates something else, please share your experience.

Here is my trophy for tonight, a Ten Micron Ribbon.
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TenMicronRibbon.jpg
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Re: New to Me American Optical 815 Microtome

#25 Post by Crater Eddie » Wed Jan 06, 2016 3:01 am

Excellent start! Good old Gulf Wax, 1001 uses.
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Re: New to Me American Optical 815 Microtome

#26 Post by einman » Wed Jan 06, 2016 4:17 am

As mentioned I believe that is a function of the melting point of the wax. Paraffin varies. You want a "harder" wax but not brittle. The paraffin you chose could be just too soft.

Leica makes a booklet in Pdf you may want to review.

http://www.leicabiosystems.com/fileadmi ... online.pdf

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Re: New to Me American Optical 815 Microtome

#27 Post by mrsonchus » Wed Jan 06, 2016 5:07 am

Hi Rod, lovely job with the knife. From your picture I can tell you a few things certainly, and all of them good;

1) Your wax is performing very well - the 'bunching' of the wax as you describe it is not only perfectly normal with sectioning but the extent to which your ribbon is exhibiting it is about 80% of what I would call the best result reasonably attainable with paraffin-wax at 10µ! It's simply caused as you rightly suggested, by the resistance of the wax to the blade's attack - this is called 'compression' and is perfectly normal - a good way of assessing the degree to which it occurs is the size of your sections compared to the size of the wax-face whence they have been cut. As sections get thinner, compression increases and the cut-sections get smaller compared with the wax-face - perfectly normal.

2) Your sections look about 70% (size) to me, easily into the 'good-zone' for a 10µ section of paraffin-wax for certain. The sections would next be 'floated-out' onto the surface of water at about 42-50 deg C and within about 5-20 seconds the wax 'stretches' or 'relaxes' back into it's original shape and size - as does (ideally) any tissue-section within it. Then the section is picked up with a slide etc...
A 3µ-5µ section for example would routinely come off of the knife at about 30%-35% of the size of the sectioning-face of the wax - and be perfectly fine after floating-out!

3) Your knife's cutting-edge looks to be pretty 'low on nicks' as the sections show little scoring or damage.

4) Your knife is at about the right angle, if not you wouldn't see these nice and minimally-compressed sections - looks just right!

All's looking good at this stage and this is surely a reflection of the super job you've made of the refurbishment, knife preparation and technique thus far - a really good job! :)

Tips, the wax-block is about the right size - but you really must make it's cutting-face a parallelogram (ideally I'd say with right-angles), especially along the top and bottom edges, which must be parallel to give a good ribbon that will come of the knife straight and not curve-to one side as the sections multiply.

If your wax is sectioning at 10µ as in the picture, at about room-temp, then it's behaving just as the 'proper' wax that I use for my sections, which has a melting point as mentioned, at about 58°C (haha - the degree-symbol at last - 'ALT-248'!). Higher mp wax generally means harder wax and cleaner sections..

The possibly tricky news is that your tissue will very likely still be 'mushy' when encased in wax and sectioned - the tissue doesn't section without being thoroughly infiltrated with wax rather than only embedded within it - unfortunately..

If you fancy 'going the whole hog' with sectioning have a look through some of my posts - it can be quite an involved process to perform well.

Good luck and really well done! :D
John B

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Re: New to Me American Optical 815 Microtome

#28 Post by rnabholz » Wed Jan 06, 2016 2:09 pm

Crater Eddie wrote:Excellent start! Good old Gulf Wax, 1001 uses.
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Thanks Eddie, this maybe one of the more exotic!

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Re: New to Me American Optical 815 Microtome

#29 Post by rnabholz » Wed Jan 06, 2016 2:11 pm

einman wrote:As mentioned I believe that is a function of the melting point of the wax. Paraffin varies. You want a "harder" wax but not brittle. The paraffin you chose could be just too soft.

Leica makes a booklet in Pdf you may want to review.

http://www.leicabiosystems.com/fileadmi ... online.pdf

Downloaded, read, and archived. Thanks einman.

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Re: New to Me American Optical 815 Microtome

#30 Post by rnabholz » Wed Jan 06, 2016 2:26 pm

mrsonchus wrote:Hi Rod, lovely job with the knife. From your picture I can tell you a few things certainly, and all of them good;

1) Your wax is performing very well - the 'bunching' of the wax as you describe it is not only perfectly normal with sectioning but the extent to which your ribbon is exhibiting it is about 80% of what I would call the best result reasonably attainable with paraffin-wax at 10µ! It's simply caused as you rightly suggested, by the resistance of the wax to the blade's attack - this is called 'compression' and is perfectly normal - a good way of assessing the degree to which it occurs is the size of your sections compared to the size of the wax-face whence they have been cut. As sections get thinner, compression increases and the cut-sections get smaller compared with the wax-face - perfectly normal.

2) Your sections look about 70% (size) to me, easily into the 'good-zone' for a 10µ section of paraffin-wax for certain. The sections would next be 'floated-out' onto the surface of water at about 42-50 deg C and within about 5-20 seconds the wax 'stretches' or 'relaxes' back into it's original shape and size - as does (ideally) any tissue-section within it. Then the section is picked up with a slide etc...
A 3µ-5µ section for example would routinely come off of the knife at about 30%-35% of the size of the sectioning-face of the wax - and be perfectly fine after floating-out!

3) Your knife's cutting-edge looks to be pretty 'low on nicks' as the sections show little scoring or damage.

4) Your knife is at about the right angle, if not you wouldn't see these nice and minimally-compressed sections - looks just right!

All's looking good at this stage and this is surely a reflection of the super job you've made of the refurbishment, knife preparation and technique thus far - a really good job! :)

Tips, the wax-block is about the right size - but you really must make it's cutting-face a parallelogram (ideally I'd say with right-angles), especially along the top and bottom edges, which must be parallel to give a good ribbon that will come of the knife straight and not curve-to one side as the sections multiply.

If your wax is sectioning at 10µ as in the picture, at about room-temp, then it's behaving just as the 'proper' wax that I use for my sections, which has a melting point as mentioned, at about 58°C (haha - the degree-symbol at last - 'ALT-248'!). Higher mp wax generally means harder wax and cleaner sections..

The possibly tricky news is that your tissue will very likely still be 'mushy' when encased in wax and sectioned - the tissue doesn't section without being thoroughly infiltrated with wax rather than only embedded within it - unfortunately..

If you fancy 'going the whole hog' with sectioning have a look through some of my posts - it can be quite an involved process to perform well.

Good luck and really well done! :D
Thanks so much for the feedback John!

It was very exciting seeing that ribbon form, what with so many unknowns. Was the machine advancing properly? Is the knife sharp enough? Is it set to the correct angle? Will the wax work? Etcetera, etcetera,

With all of that sorted, I will have to go back through all of your postings and read with a different eye. I am not sure that I have the time or patience to pursue things to the level of excellence you have achieved, but it would be a shame not to undertake to use this fine machine to at least some capable degree.

Thanks again for sharing your experience and the vote of confidence. Truly appreciated!

Rod

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