Onion root tip mitosis experiment and observation of stages
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- Опубліковано 19 тра 2024
- Onion root tip mitosis experiment and observation of stages. You’ll learn the complete procedure of onion root tip squash preparation and prepare a temporary slide in the laboratory practical to study different stages of mitosis under a microscope. An onion root tip cells are rapidly growing cells of the onion and one can observe many cells in different stages of mitosis. To observe the chromosomes of individual cells easily, the onion root tips can be prepared and squashed in a way that allows them to be flattened on a microscopic slide. This Mitosis in Onion Root tip experiment is included in the class 12 Biology syllabus. Mitosis experiment using an onion root tip demonstrates the complete procedure. It also shows how to make onion root tip squash to perform Onion Root Tip mitosis experiment". Study of Mitosis in Onion Root tip practical is based on the preparation of a temporary mount of the onion root tip to study mitosis.
It is really easy to follow these steps which I have showed in this 10 minutes video to get all stages of Mitosis in a single attempt.
For detailed information visit: www.ebiologylab.com/experimen...
Requirements:
1) Acetocarmine stain: amzn.to/3vGEZwZ
2) Dropper:
3) Slide Box and coverslips:
Slide storage box: amzn.to/3zyshnj
Slides: amzn.to/2UhMNZh
Coverslips: amzn.to/3gKxY8X
4) Compound microscope: amzn.to/2U6Lj3S
5) 90% ethyl alcohol: amzn.to/3xsJpZS
6) Sterile cotton: amzn.to/3iOKdEa
7) Sterile needle: amzn.to/2U6LuMA
8) Reagent Bottles: amzn.to/3xvRBZj
Important steps are as follows:
1) How to grow onion root tips, how to harvest and how to preserve them.
2) Choose an onion root tip for squash preparation to observe different phases of Mitosis.
3) Make sure that the onion root tip is fixed in ACETO-ALCOHOL (1:1) for one hour before you start your practical procedure.
4) Take that onion root tip on clean Microscope Slide and wash it with water.
5) Before adding any reagent or stain drain off previous excess liquid with blotting paper.
6) Hydrolysis of Onion root tip with HCl is a very important step. Follow the steps as shown in the video.
7) Staining of onion root tip cells is the most important step of this Mitosis in onion root tip practical.
8) I have used glycerin to mount onion root tip squash. Because at the time of examination your examiner may not come to see your slide before it gets dry (if mounted in water).
9) Focusing the slide under the microscope: start with 4x objective and locate the properly spread area with cells. Then focus on the 10x objective. And identify the stages of mitosis i.e. prophase, metaphase, anaphase, telophase, and cytokinesis.
staining of nucleic acid by Aceto-carmine.
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0:00 Introduction
0:25 Take two transparent glass tumblers with narrow bottom and wide mouth
1:11 Use 70% Alcohol to store roots for long period
1:23 To hydrolyse extracellular material
1:27 Use Aceto-carmine stain for cromosomal staining
1:41 Forcep to take out root tips from 70% alcohol
1:44 Use needles to place cover slip over the root tip
1:57 Razer blade to cut excess of root tip
2:23 water to clean root
2:31 take out one root tip on slide
3:05 Remove excess part of root tip
3:33 Add 2 to 3 drops of 1 N HCI
4:05 Remove HCL with the help of blotting paper to stop further effects on root tip
4:41 take out water with the help of blotting paper
4:49 Add Aceto-carmine stain over the tip
6:02 Add two more drops of stain and avoid tissue drying
6:51 Wash root tip with water and take out excess stain
7:36 Add a drop of glycerine
8:12 Cover the tissue with a coverslip
Hi, great video!! I have several questions. Are there any other alternatives to the aceto:alcohol 1:1? Can you please explain what you meant by 1:1? Also, is it a necessary step for us to use the 70% alcohol, or is it that we only use it if we need to store the roots for a longer period of time, like overnight?
Hello Jason...
Thank you for showing interest 😊....
And I love to answer questions from students... So... For your first question.... Aceto:Alcohol works as a fixative... It fixes the cell to its original state... No further changes will occur... 1:1 means.... One part glacial acetic acid and one part of ethanol... Also if it is 1:3 then one part of glacial acetic acid and 3 parts of ethanol... It means if you prepare 4 ml of 1:3 aceto: alcohol... Then there will be 1 ml of glacial acetic acid and 3 ml of ethanol.... And answer for second question.... To store root tips for longer period... As you said already... If you want to store root tips only for overnight... Then use 1:3 aceto: alcohol fixative... Instead of 1:1.... 😊👍
DK Biosolutions How does 1:3 and 1:1 aceto:alcohol affect the fixation of the root tips? Do they have different effects?
If we use 1:3 .... It will take more time to fix root tips... People use it for overnight fixation.... And 1:1 is for quick one hour fixation...
DK Biosolutions Thanks sir!! Very helpful!
My pleasure 😊👍
The music made the practical romantic 😂
I have my board practical today, this has helped me a lot in the last few hours before the exam! Thank you so much for this clear and detailed video
Hello Jehov...
I am really glad to hear that it helped you 😊👍...
Best luck for your future exams....
@@Ausmo I have used it from UA-cam... There is provision by UA-cam itself. UA-cam provide music for creator. Its copyright free music...
super bro first i cant spot the diff.phases when you given arrow mark and explained i can understand clearly
Thank you for great video Dipak Sir.
Nice video. It has standard data. Nicely presented as well.
Nicely created with caption. Thnx for creating bcz is it found to B very helpful.
Thanks! I really had trouble distinguishing the phases, but this gives me a good idea to identify the phase
My pleasure 😊 👍...
10:00 metaphase
Botany practical ... amazing explanation 💐💐👏👏👏👏👏
Great video sir 👌👌👌
Suprb..✌👌
Thank you the detailed video! This helped me a lot in identifying them
My pleasure 😊👍... Glad to hear that it helped you...
Thank you it is very helpful for us
Tq bro for your clear explanation. And your clean way of doing it with good equipment
Thank you Indra.... 😊👍
Nice one sir ✌🏻
Very great initiative towards educational documents sir, this will be very helpful for the students who are eager to learn.
We wish to see more such videos from you.
Thank you Sagar
Perfection !! Loved it..
Thanku so much sir for your guidance😊
Always welcome 😊👍... So glad to hear this 😊...
Best luck for your exams...😊👍
Very nice video sir.
ty so much I have my practical exam today and I am relieved seeing this...
Best luck for your exam 😊👍...
It is really useful information....
Thanx bro. Now I think my practical exam goes very well...
You are welcome 😊... Best luck for your exam Kishan... 😊👍
Nice video sir 👍
Best video on onion root tip till now
Loved ur work
Thank you for appreciation Girja.... 😊👍
This is what most students are awaiting.... Thank you sir.... I can use this to teach students as well...
Thank you Mel😊
Are u a sir ?
@@SAHIDGAMER yes
@@SAHIDGAMER i am a sir.....and the person who has made this video is my sir.....
Mentioning like this is very proud moment Mel... Seriously... Like a award... 😊😊😊
Nice video sir 👌👌
Beautifully done👌
Thank you 😊
Excellent
Best video for this experiment 👏🏻
Thank you Bhoomi...😊👍
this is the best video for this experiment out there ✌
Thank you 😊👍
Very nice and useful video. 👍🏾👍🏾👍🏾
Thank you Azraj Sir 😊👍...
Thank you so much sir......this helped me a lot, today is my practical exam😊😊😊😊😊😊
My pleasure😊...
Glad to hear that it helped you...
Best luck for your exam... 😊👍
it was a awesome video since at my school it was not coming clearly but after seeing your video it hopefully came
Follow the instructions, u will get the best results... 👍
Nicely and properly explained god work sir
Thank you 😊👍
Nice video sir
Superb sir . Thanks for uploading this video
My pleasure 😊👍
Thank you so much actually know I got it exactly what to do thanks alot
Heyyy Prakash...
Happy to hear that it helped you 😊👍...
DK Biosolutions really it helped me alot thank you once again
this video is very helpfull thanks bro 😀😀😀
My pleasure 😊👍
Thank u so much sir. I have my lab exam tmr and this helped me a lot in revision😊
Hello Maithry...
I am glad to hear that it helped you...
Best luck for your exam... 😊👍
@@DKBiosolutions Thank u sir😊
Nicely clear under micro scope
Thank you for considering my video..
Awesome man 👍
Thank you 😊
Great work.
Thank you 😊
Thank you sir
(Tommorow is my practical .)
Best luck for your exam Rahul... 😊👍
Same here
Best luck for your exam too... 😁👍
Same here🤗
@@mayurpankhimozinder2120 Best luck for the exam.....
So nice boss this video is very helpful for students tq
Thank you for your appreciation 😊👍...
Very helpful
Anaphase and telophase
Right 👍😊
Nice sir
Thankuh sir thankuh so much this is really very very helpful
Always welcome 😊👍...
Tommarow is my practical exam
Thanks bro😘
Hello Waseem...
Focus on staining, leave tips for 10-15 minutes in stain.
Best luck for your exam.... 😊👍
@@DKBiosolutions yes bro
Thank you
Hello. Can you help me? Is it aceton mixed with alcohol or is it formalin-aceto-alcohol, im planning to do this experiment but when i looked for this ingredient i saw that both aceton and formalin is a fixative and i couldnt figure out which one. (Chemicals are not my strong suit im sorry if what im saying doesnt makes sense)
Hello Sinan,
Yes, definitely I will help you with this. And your question is very important, to get good results. To fix onion root tips I have used Acetic acid (not the Acetone)and Alcohol in equal amount. If you have 24 hours to fix the tips then use 1:3 Glacial Acetic acid: Alcohol. Otherwise you can use 1:1 for 2 hours.
Acetone and Formalin are also fixatives but we use them for histological preparations i.e. Animal tissue section preparation.
I hope this information will help you...
Please let me know if you have any queries regarding the same...
Best luck 😊👍
Thank you! Hope you have a nice day.
Thanks, you too😊
This dam good bro
धन्यवाद 😊👍
Sir on doubt.u use after the heating on the slide another solution but our clg lab after the heating on slide use
acetocarmine stain.both are the wright???
You should heat slide with root tip and acetocarmine stain together...
is it necessary to add glycerine in slide or we can use water by replace of glycerine?plss tell me
You can use water. If you use glycerine then you can use that slide for long period.
Thank you so very much for this video. This is so helpful for my research. I just wanna ask if I can use a weaker acid to hydrolyze the onion root tip because I've seen that you use Hydrochloric acid which is very acidic and corrosive.
My pleasure 😊👍... And yes you can use low concentration acid... It should digest the extra cellular material so that you can squash that tip easily....
I tried to use acetic acid instead of hydrochloric and it didn't seem to break down the extra cellular material. So there's something I'd like to ask you as well. Is it safe to use 1 N HCl? I've seen in the video that you were not wearing gloves while handling this chemical and So can I safely use it in my experiment?
@@DKBiosolutions hope you could answer tmy question.. just want to know if 1 N HCl is safe to use..
Yes you can use it in your experiment... Diluted HCL is not that much reactive... Use gloves if you want to be more safe while handling it...
Last question.. Should I also wear goggles and face mask while handling it?
Sir, is this applicable to every 12th boards because, we don't have adding of glycerine while making slide?
It is applicable to every biology student. I have used glycerin to prevent slide from getting dry. You can use drop of stain. But seal your coverslip to slide using nail polish.
Nice video sir 👍👍
Thank you Sahara... 😊👍...
I have a question, can i use safranin instead of acetocarmine? is safranin good enough for observing mitosis?
Yes you can use it... It stains nucleic acids...
In onion root tip mitosis slide preparation except aceto cermin which re agent can be used ?
You can use aceto-orcein Toluidine blue 0.05% , Feulgen stain instead of acetocarmine, Basically any nuclear stain...
Thanks bro
My pleasure 😊👍
superb experiment
Thank you 😊👍...
Very nice sir :)
Nice
Sir thank very much
My pleasure 😊
Wow
Super explanation sir
Keep watching
thanks a loads
My pleasure 😊👍
sir hamy root grow krwany ka just one day mila hai to hm 12pm tk grow krwa skty hein aj or kal tk grow ho skti hai?or sir thank you for guidance,,or ham kisi b place pr rakh kr root grow krwa skty hein?just ek root b ajaye to thik hoga
Hello...
1 din me possible nahi hai... onion ka base jo hai o bas water ke contact me aana chahiye... 3-4 days lagenge atleast... Aap try kijiye... Growth hormone bhi hote hai magar aapko available honge ki nahi pata nahi... Usase jaldi aa jate hai...
Well explanation
Thank you 😊👍
If I want to count the chromosome number , can I use this technique?
To count the number of chromosomes, you have to grow onion roots in chemical like cholchicine (which does not allow the spindle microtubule to form). Thus all cells get arrested at metaphase. Then cells are to be given treatment of hypotonic solution (so that cells get turgid). Such cells are to be thrown on chilled glass slide, so as to break them open and you get chromosome spread. These chromosomes you can stain and visualised in microscope (oil immersion). This will give you a rough count of chromosomes. The best way to do chromosome counting would be to use flow cytometer. Cheers,😊👍
Can we use acetocarmine to dye human chromosomes as well? Or is it suitable only for plant cells?
Yes, we can use it for animal cell chromosome staining... some people use aceto-orcein stain to stain chromosomes of animal cells... But the protocol is different than this staining procedure...
@@DKBiosolutions Thank you veryyyy much!!
My pleasure 😊👍
Super
Tq sir nice video, 🙏
My pleasure 😊👍...
Thanku sir🙂
Welcome 😊👍...
Can i use Safranin stain instead of Aceto-carmine stain?
Yes, you can 👍
IS it possible to use methylene blue stain instead of aceto-carmine stain and use the same mechanism?
Yes... You can use 1% methylene blue instead of acetocarmine to study mitosis in onion root tips... But you have to standardise the mechanism 👍😊
@@DKBiosolutions Thank you for the answer. could you please tell me what does standardising mean? Can't I repeat the same steps as in this video?
My pleasure 😊...
Standardise means, if you want to use methylene blue instead of acetocarmine then, you have to repeat experimental procedures till you get the good results. While repeating make some small changes in staining protocol... 😊👍
@@DKBiosolutions thanks❤️🙏
My pleasure 😊👍
Is it compulsory to heat with HCl
Or we can heat with water?
You can heat with HCL or Stain
Thanks
Thanks
Welcome Basavaraj...
What other stain can be used in place of aceto-carmine? Kindly reply asap 😊😊
Hello Sunita...
You can use aceto orcein stain instead of aceto carmine. ua-cam.com/video/YdXO0FTgvSY/v-deo.html this video shows comparison with both the stains. Best luck 😊👍...
@@DKBiosolutions i didnt find both of these stains...can i use methylene blue?..
Yes you can use it too...
Methylene Blue is a synthetic basic dye, stains to negatively charged cell components like nucleic acids...
Why are we using acetocarmine stain instead of methlene blue or any other stain???
Acetocarmine is a chromosomal stain... It binds and stains negatively charged components of cell like DNA and RNA... same with Methylene blue, also known as methylthioninium chloride, can be used to stain or highlight parts of animal, bacteria, and blood tissue specimens in microbiology. It also binds to negatively charged components of cell with low affinity. For mitosis we use acetocarmine or aceto-orcien stain....
Sir can you please explain how to grow the onion quickly...I already kept in my home the onion with tumbler but 5 days passed still the tip is not growing...so what to do??
Add very small amount of npk fertilizer...
Oky sir I will try... Thank you soo much sir☺️👍👍
My pleasure 😊👍
I think in the first pic, there're in Telophase ...
Can u tell me plz how i can prepare 3:1 aceto: alcohol solutions
Sure... Mix 3 ml glacial acetic acid + 1 ml ethanol... It will become 4 ml 3:1 aceto: alcohol...
Thank u
My pleasure 😊👍
Glycerin ki jagah kya add kr skte hai...plz bataiye
Glycerine use ki jagah water bhi use kar sakte hai. Glycerine agar use karte ho to slide dry hone ka koi chance nahi, water kuch der bad evaporate ho jata hai. Exam me ye bat problem create karti hai.
@@DKBiosolutions thank u so much for replying me...its is gonna useful to me thanks again...😃😃😃😃
oil can work
Hello Durgvijay...
We can't use oil as a mounting media. Oil is used to focus cells under higher magnifications. But to check whether it works or not, you can experiment on it😊👍.
This is what we want before a day of exam 😄😁😃😀
Best luck for your exam.... 😊👍
@@DKBiosolutions thanks sir actually again replaying your video so that I perform my best😌
😁😊👍
In practical exam if we are not able to identify the stage which stage should I write?
Hello Kochumon...
You can write and draw prophase stage... Because most of the cells are in prophase stage...
Use freshly prepared onion tip for slide preparation... Locate the tip end after squashing it... And try to find other stages of mitosis... Best luck 😊👍
Tomorrow is my practical exam do you have any video related to saliva experiment
No, I don't have any video related to saliva... Anyway, best luck for your exam... 😊👍
Super sir
Thank you Chandru... 😊👍
Hello, may i ask what glycerine is for?
It used to keep your root tip spread as it is for long period... It prevents drying...
@@DKBiosolutions thank you. Your video is very helpful!
My pleasure 😊👍...
Arigatogozaimashita😭😭😭😭😭thank u so much.....
どういたしまして 😊👍...
@@DKBiosolutions 😱😱😱
😁😁😁 グーグル翻訳
@@DKBiosolutions 😑yeah m having that too...but it took me 5 minutes to write it..in my handwriting on that app whatever u said.I actually thought u knew it😏
I was curious about your first comment, so just translated it and found that in Japanese language. And replied in same language. 🙂
can I use another dye?
Yes you can... But it should be chromosomal staining...
@@DKBiosolutions thanks
can we use something else instead of acetocarmin
You can try... It should stain chromosomes...
Telophase and anaphase
Metaphase and telophase
Am i right?
Yes, you are right 😊
no u are wrongt
Please guide us😊👍...
Explain how it is wrong. I am agree with you, its not telophase. But explain us how it is not telophase.
DK Biosolutions because in telophase chromosomes reach the opposite pole lose their individuality and look like a mass of chromatin
Nuclear membrane appears to form the nuclei of two future daughter cells
@@saiyadfiza5934 absolutely right explanation, this is what an examiner expects from students. I felt really great after reading your answer...😀👍
This is a best video
How to find the tip of the onion root? Please tell me 🙏
Hello Himadri...
Before staining the Onion Root tip you can see a blunt end and a sharp end made by cutting with blade. We need to focus on blunt end of root tip to find the mitosis phases. After staining you will see that blunt end is darkly stained than remaining part. 😁👍...
@@DKBiosolutions Thank you very much 👍✌
Sir your roots were already present in the fixative as you haven't put it in fixative???
Yes... You are right... roots were already in fixative...
@@DKBiosolutions so sir if school will give us roots in any solution that solution is of fixative???
Most of the time they provide you roots in fixative only... Because fixation takes time... From 1 hour to 24 hours... So ask your teacher before they provide you the root tips...
@@DKBiosolutions okk sir thanks and sir can we warm acetocarmine after applying coverslip
@@Gamya-zo6cf yes... It can be done....
I m a teacher, and its my first of teaching 12th biology , so plz help me about practicals, that how and how many experiments , i hv to do for the students...
Hello sir...
I don't teach to 11th and 12th... I don't know much about it. I teach to B.Sc students...
I feel bad, I can't help you with it... 🙏
@@DKBiosolutions its all right, thanku for the precious reply
My pleasure 😊👍...
Metaphase and telophase
Right 😊👍
Hcl dalne ke baad heat nahi karna kya
HCl ke sath heat kar sakte hai, fir stain ke sath jyada warm nahi karna padega
@@DKBiosolutions; Okk thanks
Is it Anaphase ??
Yes... Anaphase and on next image Anaphase and metaphase...
Best luck for your exam.....😊👍
What's aceto-alcohol?
Hi...
We have used Aceto- Alcohol as a fixative. To fix onion root tips I have used Acetic acid (not the Acetone)and Alcohol in equal amount. If you have 24 hours to fix the tips then use 1:3, Glacial Acetic acid: Alcohol.
@@DKBiosolutions thankss
hello what music is playing, it just hits different
Hi... It was available in the software which I have used to edit this video 😁
@@DKBiosolutions what software do you use?
Camtasia
@@DKBiosolutions thanks bro B)
Welcome 👍
Sir agr koi b phase nhi aara ho to
Yessa thabhi hoga jab root tips acche nahi honge, otherwise dikhna hi chahiye... 2-3 tips ek sath process karna. Dhyan rkhana ki tip ka end kis side me hai, wahi phases find karna...