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Introduction to microtomy
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- Опубліковано 14 сер 2024
- The ability to cut sections from paraffinised blocks of tissue is a routine procedure within pathology laboratories and a valuable skill for research histologists. During a break in teaching I managed to cut a few decent sections. Experts will no doubt see the flaws in my technique, but this video demonstrates the basics for those looking to get started.
This is the most realistic demo ever! It really helped me on my online class in histotech. Thank you so much! Hoping for your good health!
As a medical lab student who was lost.. Wanna say thank you lots ❤❤
You are most welcome Hannah. Let me know if there are any other topics that you would like to see covered.
The online course is also still available for Trainee Histologist level members and above.
I love the way you daletailed possible scenario in histopath. Thank you Doctor. Such a wonderful learning just by watching this due to pandemic we lost the chance in having actual pathology lab sessions.
Am glad to help!
Thank you very much for this demonstration, as a new Microtomist, it’s quite encouraging to see you problem solve and persevere when the blocks aren’t cooperating. Thank you for all you do ☺️
You are welcome!
Really good real world example of sectioning, well done & thanks for including examples of typical issues/problems in taking sections, the door left open was a great touch ..
Thank you Damien for the fantastic demonstration and explanations! I thought it was really cool that you used a POV camera angle to show us exactly what we could expect to see during the process. Just earned yourself a sub!
I´m so thankful with the way you made this video. I´m just starting to work with the microtome and had a lot of issues with my sections cause they turned into rolls and things like that. It´s kind of recomforting to me to see that those are things that are more than common and can happen at any time. Also all of your tips and your technique will help me better mine. Thank you very very much for this video.
Thanks Hans. Am pleased to hear that the video was useful.
OMG thank you so much!!!! My sections were clumping like that and I did not know why and I kept practicing for days like that.. I will try now with the cold block... and also I was scooping with the slide hahaha..It was so stressful. I will try with these tricks now.
Glad to help!
Let me know how you go with colder blocks.
As a laboratory student when i found your videos it meant the world to me they are sooo sooo soo helpfuul thank u soo much for the efforts ❤❤❤
The channel is made for people like you so I’m glad that you’ve found it useful. Please share your experience with others so that I can grow the channel.
If at first, you don't succeed try, try again! Really enjoyed this.
Yes, I was glad to be able to capture the full experience of frustration and eventual success.
I'm in healthcare IT and was looking to understand the process of histopathology better. Thanks for this explanation.
Watched from Uganda in preparation for histopathology exams. Thanks alot for the great job.
My pleasure Peter
Good luck with the exams!
Siiba bulungi
I didn't know you could pick up sections in the middle of a ribbon like that!! I've been struggling to separate each section with tweezers while they're floating on the water before picking them up on the slide. I'm going to try that tomorrow. 😊
This was so practical and I really loved it and have really learnt a lot thanks so much
Glad it was helpful!
Excellent. Good to see a demo with "warts and all"
Glad you liked it!
Very informative explanation Prof damien harkin .Thank you for all you do
You are very welcome
@@damienharkinsir please tell me the temperature of water bath ??
Appreciated this advice about cooling the block - 3 years into a PhD project and the tip had never come up *sigh* - hopefully improved tissue sections to come.
Btw safety first! Should definitely have those gloves on around the microtome blade 🔪
Thanks Christopher!
Good luck with the PhD!
Thanks for raising this point.
It’s been raised before and I’ve yet to really respond properly.
The microtome blades are certainly very sharp, and so every care should be taken. The safety measures that we employ are generally sufficient to reduce the risk of a cut. Wearing latex or nitrile gloves may well reduce risk further, but the experienced microtomists that I know, don’t wear them.
The tissue itself should be safe as has been fixed in formaldehyde and embedded in paraffin, but we do recommend wearing gloves if sectioning brain or spinal cord since prions (e.g. CJD or vCJD), if present, remain infectious.
Great video! very informative and helpful
Thanks so much for your sharing, it helps a lot!
This block is so good.. How to make this block??
Excellent Demo, thank you sir
You are most welcome.
Please consider supporting the channel as a member for access to more material.
That's a really nice microtome! Much nicer than that little round thing they sell on Amazon. I bet that one costs a few hundred buckazoids!
I would have liked to see how you embedded the sample in the wax block.
I demonstrate embedding of tissue in paraffin in this video.
Overview of techniques
ua-cam.com/video/8Rv7Gx_4xqQ/v-deo.html
Could someone please advice on how to position the knife so that is cuts the tissue well?i struggle with positioning the knife which was not touched upon much?
Really nice and clear video, thank you a lot!! :)
Hi Ivana,
There are multiple factors to consider here starting with the specimen itself.
During embedding, the tissue should be oriented such that it display least resistance when passing the blade. For example, for skin, cut from dermal side first and epidermis last.
The angle of the block face should match the blade and there is usually a recommended angle mark on the blade holder.
The knife holder can usually be moved sideways to enable a fresh area of blade to be used when cutting best quality sections.
You should also cool your block if excessive crumpling occurs.
I hope that helps a little.
🤓
Thanks for this video. Why do we need to bake the slides in the oven after mounting?
You need to bake in order to ensure good attachment of sections to the slide.
Thank you
You're welcome
Thank you for the go pro video.! one suggestion though - gloves.
Cause of ribbon of consecutive section are curve
Thank you 😊
You're welcome 😊
Thank you, this video was really helpful
My pleasure. New videos coming later this year.
Would love to see more videos from you.
Thanks Jennifer. More videos coming soon.
I try not to be greedy but I need to make many cross sections of flower which takes about 6 hours and I sometimes loose most parts of it
We’ve all done this at some point but then realise that patience is essential.
Your blades are too sharp. That's why they are rolling up on you like that. Take a kim wipe and gently glide it across the the blade to dull it slightly and you'll find they won't roll up like that.
Thanks for the tip Will. I'll give this a go and let our students know as well.
a lot of hot air from talking doesn't help' I remember holding my breath handling ribbons in micro-tech. A was quite patient for the temperature information sought but to no avail. What was the cutting block's desired temp you were trying to achieve?
Great question John!
We use a chilled cold plate or block of ice to cool the block over approximately 5 minutes after facing the block.
The block will then warm up when mounted onto the microtome and is probably around 10-15 degrees C when sections are being cut but I have never directly measured the temperature.
Maybe i missed it I think it is quite important to talk about the cutting angle of the knife. Make sure it is set properly
Yes, good point. Have been considering developing another video to discuss these issues, including adjusting the block orientation in the chuck when required.
@@damienharkin That would be great! I've still got some problems with adjusting the blocks, so it would be nice to receive some tips :)
Try gently blowing on the ribbon as you section. It should prevent creases from happening :)
As a layman; does this risk cross contaminating the samples?
How do you prevent bubbles in the tissue when draping sections into the water bath? Is there a technique used to get rid of them, or a solution to use? Thank you!
Hi Adrienne, yes, bubbles can be a hassle. They tend to form as the water bath warms up. Once bath has reached desired temperature, they can usually be wiped way using paper towel.
Is there any hand microtome that isn't so expensive?
Sir please tell me what is the temperature of water bath ???
Between 40-50 degrees C.
@@damienharkin Thanq so much sir ❤️🙏
Hi sir,do you leave slides to airdry or you use slide dryer
Hi Fatma,
The slides need to bake in a drying oven for around an hour at 60-70 degrees C in order to stick the tissue to the slide.
You need to bake the slides in a drying oven at 60-70 degrees C for 45-60 min
Can you please do a Pentachrome stain?
Will do. I have the raw footage for Movat's PentaChrome. Just need to put it together. Hope to post within the next few weeks.
Damien Harkin Thank you! Will be looking forward to it.
Pencils come to save the day.
Really just here cause my lecturer added a link 😢😢😭
Say hello to your lecturer for me!
K now I'm clear he have camera on his head.