Just FYI peeps. Certain tissue types need more hydration ( hence the variance in soaking times). If a tissue is dry for sectioning such as brain or liver, it needs to sit on wet ice for longer. If tissue is fatty such as deep melanomas, breast, lipomas etc soaking the block on water is about the biggest mistake you can make. It will interfere with sectioning if you introduce water after processing. It will be incompatible with xylene that is still present in the tissue and explode on the water bath. The whole point of soaking your block prior to sectioning is to hydrate the tissue so that the morphology is more accurate on the slide.
I´ve never heard about soaking paraffin embedded tissue samples for 24 hours in ice water...I´m in histology for 20 years now....also it is very dangerous trimming a paraffinblock over a sharp knife without gloves.....
That’s the most ridiculous technique for floating ribbon. Why wouldn’t you just anchor your ribbon the one side of your water bath. Who wants to chase the ribbon all over their water like that? You must work in a very low volume lab to be soaking your blocks for one to twenty four hours. That’s insane!!! Who has processors that are drying the tissue out that much they need soaking for hours?
Me gustaria que me ayudaran. Tengo el mismo modelo de microtomo y últimamente cuando estoy sacando la tira siento que el microtomo da pequeño salto entonces sale un fino y uno grueso uno fino y uno grueso. Por.lo regular corto en 4 micra y la.grasa en 5. Revise el microtomo y al.parecer esta calibrado. Que puede ser?
HI thanks you very much for the video. I want to aks you for a favor. Since English it's my second language I didn't get why did you put the block in water for 24 hours? Thanks again, have a great week!
Hi, I have a question. Which block mailer did you use? do you have the link where you buy it from? I couldn't find the correct block mailer to hold my samples into the microtome machine, the ones I found online were too big.
Slt, merci pour la vidéo mais j'aurais bien aimer voir la suite question pour fait fait une comparaison ! Moi je fais l'étalement avec un liquide qui est un mélange d'albumine et glycérine et eau, j'ai pas mal de fois essaye la méthode du bain marie mais après coloration les fragments se décollent , g hâte de savoir comment vous faites le deparaffinage après étalement et la coloration que vous utilisez, merci infiniment de me répondre !
Wow. 1. Who sections paraffin normally at 10um? 2. You soak the block on the chuck. 3. Use a god damn brush to align the sections on the slide! Does no one look at histotech manual (Armed Forces) anymore?
@@diatomsaus So I haven't checked out any histo videos on youtube in forever because there just wasn't anything good but that could have changed by now. The best way to learn is to watch experienced people that you work with and ask questions. Honestly microtomy is about practice and muscle memory. if you are having specific issues check out ihcworld.com's troubleshooting guide. Just google microtome sectioning troubleshooting.
@@towens1168 Oh wow, thanks for the information! I didn't expect a response on a 2-year-old comment. Nice suggestions there. The lab I work at doesn't deal with sectioning at all, there's a biology one that I don't have access to, they do all the sectioning. I guess I'll try and somehow wiggle in to learn stuff. I'm looking to grow my personal "lab" for microscopy imaging rather than anything professional. Sometimes, these manual microtomes pop up on flea market sites at alright prices.
@@diatomsaus Yeah no problem. So if you're going to be using already prepared blocks that might work. You will still need to buy the chemicals to H&E stain the slides at the very least. I don't think Hematoxylin or Eosin are controlled chemicals so you should be able to get some if you want (I could be wrong about that never looked them up). If you want to prepare your own blocks that gets much more complicated. You need to fix the tissue first and then process it and then you need to be able to embed it. I imagine that could all be done without a processor or an embedding station but it would be a pain at best. Processing takes anywhere from a few hours to overnight. Honestly there's a lot of work and money (and skill) that goes into making a slide. If the imaging is what you're really interested in you might be better off just buying slides.
@@towens1168 Yeah, at the moment I have a big collection of slides. Some vintage, some from preparers such as Klaus Kemp who deals with diatoms, recently (sadly) retired. Out of all the Chinese ones I've tried, I found one university ran company that offers great quality at prices that are a lot higher (similar prices to western companies), but quality is what I'm after so it's fine. Generally, Carolina(dot)com offers exceptional slides to anyone who wants to buy prepared ones, their shipping costs isn't bad either. I've already spent enough money on all the microscope stuff. I've decided to forget about making my own slides unless I get the chances to do so at work (our labs have pretty liberal policies, we can visit for our own projects luckily). A microtome is anywhere from $500-$1000+ with blades being another expensive disposable, along with all the staining chemicals and issues with dust/fibres which sort of warrants a benchtop clean-room assembly cabinet... it's an endless hole. Either way, thanks for your insight!
I don't even know where to begin! There were so many mistakes here its not even worth trying to call them all out. I'm sure there are some good videos but every one I have seen on UA-cam sucks. Will someone actually good at there job make a video on a sectioning. This is so cringe worthy!
Just FYI peeps. Certain tissue types need more hydration ( hence the variance in soaking times). If a tissue is dry for sectioning such as brain or liver, it needs to sit on wet ice for longer. If tissue is fatty such as deep melanomas, breast, lipomas etc soaking the block on water is about the biggest mistake you can make. It will interfere with sectioning if you introduce water after processing. It will be incompatible with xylene that is still present in the tissue and explode on the water bath. The whole point of soaking your block prior to sectioning is to hydrate the tissue so that the morphology is more accurate on the slide.
I have been taught to keep blocks on ice after trimming. Sections just fall off at 4microns
"put the block in water, after 24 hrs put the block back..." lol homie I got to finish sectioning so I can go home
Same 😂
Never heard of that before.. a couple mins should do the job
please always, ALWAYS!!! cover the blade (and lock the flywheel) when bring your fingers near the blade - Histotech Instructor
if you do that though now days you will be called slow ext ext
You make it look so easy!
Best video I have found yet for refreshing my knowledge on sectioning, thanks!
It’s not reallyeasy to use
Excellent. It is a detailed explanation.
I´ve never heard about soaking paraffin embedded tissue samples for 24 hours in ice water...I´m in histology for 20 years now....also it is very dangerous trimming a paraffinblock over a sharp knife without gloves.....
I do put the blocks while trimming in ice water when its hot, it prevents from "melting" a bit
@@melaniee467 how to trim...?
This video absolutely gives an knowledge about the microtome cutter too...........
2:45 Why do you have to remove it and place it in water for 24 hours when the tissue is exposed? Does it help the blade cut better into the tissue?
That’s the most ridiculous technique for floating ribbon. Why wouldn’t you just anchor your ribbon the one side of your water bath. Who wants to chase the ribbon all over their water like that? You must work in a very low volume lab to be soaking your blocks for one to twenty four hours. That’s insane!!! Who has processors that are drying the tissue out that much they need soaking for hours?
What type of tissue are u sectioning that you soak for 24 hrs? Is this for special studies or routine staining?
I got the same question.. why do you have to place it in water for 24h after the tissue is exposed?
It should be 10% alcohol
Me gustaria que me ayudaran. Tengo el mismo modelo de microtomo y últimamente cuando estoy sacando la tira siento que el microtomo da pequeño salto entonces sale un fino y uno grueso uno fino y uno grueso. Por.lo regular corto en 4 micra y la.grasa en 5. Revise el microtomo y al.parecer esta calibrado. Que puede ser?
HI thanks you very much for the video. I want to aks you for a favor. Since English it's my second language I didn't get why did you put the block in water for 24 hours?
Thanks again, have a great week!
OMG. You do not put your fingers in the water bath or pick up sections like that.
Thank you so much! Learned a lot.
Thank you for the video as I am still a student in histology but from what I have been thought you must always have the knife guard on if not in use.
Ik it's been 5years but how has things been for you
this is good ❤️💁🏻♀️
Hi, I have a question. Which block mailer did you use? do you have the link where you buy it from? I couldn't find the correct block mailer to hold my samples into the microtome machine, the ones I found online were too big.
que modelo es el microtomo?
Wear gloves! That is lab specimen processing 101! Universal Precautions is a must when handling blood and tissues.
It's fixed, chill...
If anything, wear gloves so you don't contaminate your sections but only a few cowboys use their fingers in the water bath
He is handling fixed, dehydrated, xylene-treated, embedded blocks. He is not handling blood or fresh tissues in the video. Stop the safety paranoia.
why do you cut wax from the block??? it will be removed before colorations anyway...
wow great video! ❤
عاشت ايدك👍🏻
very useful.. thank you...
Why you put the block in the water and waiting 24hourse
Slt, merci pour la vidéo mais j'aurais bien aimer voir la suite question pour fait fait une comparaison ! Moi je fais l'étalement avec un liquide qui est un mélange d'albumine et glycérine et eau, j'ai pas mal de fois essaye la méthode du bain marie mais après coloration les fragments se décollent , g hâte de savoir comment vous faites le deparaffinage après étalement et la coloration que vous utilisez, merci infiniment de me répondre !
Wow. 1. Who sections paraffin normally at 10um? 2. You soak the block on the chuck. 3. Use a god damn brush to align the sections on the slide! Does no one look at histotech manual (Armed Forces) anymore?
Paraffin block's kept in the water for 24 Hrs. Using of water is cold or normal water??
but why
Amazing
Thank you!
Thank you very much! Very useful.
Bien. Wonderful
Thank you 😊
thank you
So nice
Can write a procedure about sectioning
very good
good Info
good thank you
doesnt it waste more time?
Very Good ...
very useful... well explained...i appreciate...
+kanaka moka I dont
sir, can we use this for cutting metal sheets??
NO. duh
Well, it's not how I did it, but I guess different strokes for different folks. Too slow, too thick, too likely to cut yourself, etc. etc.
nice
Работягам из 304 группы привет, остальным соболезную...
good
dédicace à la fac de poitiers, tous les shegueys de la nezo aie aie aie on va l'avoir notre l3 les frr
hey nice cuting thanks
What in the world are you doing!?
I don't want to be mean but please dear god if you are new to histology DO NOT use this video as a tutorial!!
What's a better guide or source for beginners? Reading the several comments here got me scared.
@@diatomsaus So I haven't checked out any histo videos on youtube in forever because there just wasn't anything good but that could have changed by now. The best way to learn is to watch experienced people that you work with and ask questions. Honestly microtomy is about practice and muscle memory. if you are having specific issues check out ihcworld.com's troubleshooting guide. Just google microtome sectioning troubleshooting.
@@towens1168 Oh wow, thanks for the information! I didn't expect a response on a 2-year-old comment. Nice suggestions there.
The lab I work at doesn't deal with sectioning at all, there's a biology one that I don't have access to, they do all the sectioning. I guess I'll try and somehow wiggle in to learn stuff. I'm looking to grow my personal "lab" for microscopy imaging rather than anything professional. Sometimes, these manual microtomes pop up on flea market sites at alright prices.
@@diatomsaus Yeah no problem. So if you're going to be using already prepared blocks that might work. You will still need to buy the chemicals to H&E stain the slides at the very least. I don't think Hematoxylin or Eosin are controlled chemicals so you should be able to get some if you want (I could be wrong about that never looked them up). If you want to prepare your own blocks that gets much more complicated. You need to fix the tissue first and then process it and then you need to be able to embed it. I imagine that could all be done without a processor or an embedding station but it would be a pain at best. Processing takes anywhere from a few hours to overnight. Honestly there's a lot of work and money (and skill) that goes into making a slide. If the imaging is what you're really interested in you might be better off just buying slides.
@@towens1168 Yeah, at the moment I have a big collection of slides. Some vintage, some from preparers such as Klaus Kemp who deals with diatoms, recently (sadly) retired. Out of all the Chinese ones I've tried, I found one university ran company that offers great quality at prices that are a lot higher (similar prices to western companies), but quality is what I'm after so it's fine. Generally, Carolina(dot)com offers exceptional slides to anyone who wants to buy prepared ones, their shipping costs isn't bad either.
I've already spent enough money on all the microscope stuff. I've decided to forget about making my own slides unless I get the chances to do so at work (our labs have pretty liberal policies, we can visit for our own projects luckily). A microtome is anywhere from $500-$1000+ with blades being another expensive disposable, along with all the staining chemicals and issues with dust/fibres which sort of warrants a benchtop clean-room assembly cabinet... it's an endless hole.
Either way, thanks for your insight!
I don't even know where to begin! There were so many mistakes here its not even worth trying to call them all out. I'm sure there are some good videos but every one I have seen on UA-cam sucks. Will someone actually good at there job make a video on a sectioning. This is so cringe worthy!
Please go and do your own video, I'm curious and want to learn
O God pray on Mohammed and the family of Mohammed the pure good and hurry their release Oh generous
good