Gram Staining

[@poojapareek1386]

We can learn this with a sentence Come In And Stain C- Crystal violet I - iodine A- alcohol ( ethanol) S- safranin

[@sendmemoneythanks]

Pooja Pareek yo thank you!! Have my micro practical tomorrow. Im a Pharm d student in Pakistan 😊😊

[@adnanahmed7463]

Thank you for helping me remember this:)

[@nocmemer4686]

Pooja di thx,I have exams tomorrow it will help me

[@poojapareek1386]

@@nocmemer4686 all the best for your exam dear😊...and no need of thanks dear I'll be great full if I can help you 😊😊

[@nocmemer4686]

So sweet of you😇

[@roselewis7683]

this is one of the best representations of a gram staining procedure I have every seen. Simple and straight forward with all the precautions built it. Well done.

[@raykyogrou0358]

Interesting! Never had to do this before, kinda wish I did. Only disappointment was at the end because I was kind of expecting to see what that slide looked like under the microscope.

[@messilionel12345]

Yeah...Me too disappointed......

[@ProfRoofs]

Here you go ---> kzbin.info/www/bejne/m5jTi2N4ma-Ee5o

[@nadaahmed6236]

@@ProfRoofs THANK YOU SO MUCHHH

[@nafisaeiramnoor3428]

I have microbiology exam next week. This video is so helpful for students of medical schools like me. Thank you for making it👏

[@sugarcoder]

Thank you for making this video! My lab partner hogs all of the lab activities and doesn't care about other people, so I could not learn it well sadly. There is a lab exam on Gram staining, so I am glad that you made this video so I can learn how to do it!

[@blackberrykathryn100]

Staining is to improve the contrast within a cell to show the organelles in greater clarity and detail. The cytosol is transparent there is little if any contrast. Using positively charged dye it attracts to the cytosol within the cell and you can distinguish between the different organelles. Using a negatively charged dye it repels the outside of the cell and stays outside the cell and distinguishes the cell from the background. The positive dye could be Methylene blue and the negative dye could be Congo red. Differential staining is used to differentiate between two different organelles or organisms. Gram stain technique is used to show gam positive bacteria and gram negative bacteria whilst the acid fast technique is used to show mycobacterium form other bacteria present.

[@jellykin7161]

Nice video! I wish I had seen this *before* doing Gram Stain for the first time as a high school student. Instead, I had to rely on an ambiguously worded worksheet for instructions. I ended up washing my slide completely clean of all cells... That was spectacular, I suppose. ah ha ha :)

[@stookiegrace1253]

Joules Kin same bit somehow my e. Coli cane out gram-positive which was correct!

[@sajanvincent1111]

Thank u for the information

[@loeyexo1685]

thank you for this. i hope i'll do well on my practicals tomorrow 😨

[@gargimahajan2273]

ᄏᄏ아라 So, how did it go?

[@firephoenix8192]

I have praticals tomorrow as well

[@iqrailyas2097]

i have practical tomorrow too

[@hollistercal888]

same lol

[@rvsingh4538]

Mine too

[@haleyknedeisen8108]

I have a gram stain lab tomorrow and this was really helpful!!

[@_Dr.P_]

I have my practical exams tomorrow And today I am watching 😂😂 So thankful to you for explaining each and every step perfectly Hope I will do well tomorrow 😂😘❤️😂

[@A9poj]

armyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyyy

[@_Dr.P_]

@@A9poj ARMY 💜💜💜💜

[@himanshurao8668]

@@A9pojstupid armyyy

[@haninasofeafaizal3600]

nooo i wanna see the results :'(

[@srija_7890]

😭

[@KevinRaizada]

Me too

[@arian138]

Thank you for saving my time, not to see the video. You lost but you saved many.

[@blackberrykathryn100]

So gram staining is to differentiate between gram positive and gram negative bacteria and acid fast technique is to differentiate mycobacterium from other bacteria

[@JessM20_]

Kathryn Russell yup!

[@firozahmad1653]

Acid fast different iate Mycobacterium tuberculosis. And m. leprae

[@BlackFlagHeathen]

Ugh I hate lab practicals. These KZbin vids are a godsend!

[@fatmaahmed832]

Thank you for this! you saved me from the embarrassment of asking my lab partner who likes to gatekeep information to himself

[@irregretableyblessed2581]

quick and straight to the point explanation

[@MelomaniacEarth]

I did it yesterday in my internship. I'm so happy.

[@SurvivalSquirrel]

Where is the observation?

[@MasterMicrobiologybyDrAdetitun]

This is great demonstration of an important concept and very concise. Weldone

[@reflexoplays6451]

we wash the slide with a bit of detergent before use to make it grease free which helps alot in spreading the water droplet and giving a smear of bigger surface area.........idk if it's right but it's a mandatory step in our university and it has become a part of staining procedure

[@Samuitsuki]

Rinsing in a beaker seems a lot more controlled than using spray bottles as they seem to do in the lab I'm taking classes in.

[@charlottemcgowan4275]

Ok, this is so different from how my lab TA did it but makes so much more sense.

[@KeyannaMoss]

Can you post all your lab experiments...lol I learned so much through this THANKS!!1

[@Kidsent567]

If it is time i will post it dr

[@niqabyousafzai6525]

thankyou, its all cleared. having exam of Gram staining tomorrow 🙌🏼

[@Whatif138]

Bestest video short and sweet 😊 staining of gram negetive bacteria 😌

[@YoyoBear12]

loved it except for the smear. it was a little too cloudy in my opinion.

[@erikhernandez6450]

i think it may have been agar residue

[@jansherkhan5835]

Awesome. Passionate to perform this on lab.

[@scholasticasonia3806]

Thanks I've understood the technique properly thanks to this video.

[@Jeene269]

My histology exam after one day! This vdu was so informative thanks ✨

[@nursepotassium]

I have a gram stain skill exam in a few days…can’t wait lol

[@yadneekadam5833]

The video was nice, but I have a question, u marked a circle with wax pencil , and made a smear on the same side???? As I'm a new student, my proff taught me to mark the slide , invert it n prepare a smear on inverted slide.... Why y did so?????😶

[@twilighteclipse7337]

Oh! This is the greater Graim Strainig procedure I've ever seen. It's so interesting.

[@edidiongekpo]

Thank you so much, I learnt a lot from this video!

[@Cccvgd13]

Thank you so much ,I learnt alot from this video!

[@khgnew763]

at very fast if you add drop of water to bacterial colony, the cells may swell and lyse due to osmotic gradient. better use a drop of isotonic normal saline

[@souravsupakar6858]

That was so interesting I am not a science student but it was like I can understand everything by watching this video

[@Notsowell123]

haha I was waiting for what it looked like under the microscope

[@joybulan1800]

Thank you helped me for my bacteriology subj

[@Krunchyz]

Shouldn’t the alcohol be continuously added, drop by drop on the smear for 30-45 seconds?

[@raphaelleteissedre7615]

I'm here because one of my character is a scientist and I wanted to describe part of the procedure for Gram straining!

[@jelenajeca4585]

This is amazing, so interesting, I like it!

[@jhoannadipay5594]

Thank you for making this video!

[@nalabagam544]

What are the other counter stains other than safranin can be used ?

[@mehboobvlog293]

Full prepared before 1 hours exam❤😁

[@NobleTenz]

wish me luk on my practical :)

[@deemmuteb447]

What's the benefit of using a wax bencil ?

[@littlesneets8026]

i understand this video is 10 years old, but i just want to be sure. is the gram decolouriser Ethanol solution? because i'm sure that's what we used for our gram staining methods to rinse the stain, but maybe i am just mixing up the wrong reagents

[@zebullon99]

Very clear and informative tutorial.

[@sulimankhan621]

good

[@eshansharma7788]

Video maker- Today, we are out of microscopes but we were in a hurry to make this video

[@destinybasedoll]

how long does it take to perform and analyze a gram stain ?

[@heyitsrayrui]

Like 15-20 mins?

[@cosmo2saucyyk630]

very well explained

[@pamelabitar9558]

If the stain of crystal violet came inti my hand how do i take it off

[@elizabethowairu9296]

What happens if you do not heat fix the bacteria? is it detrimental to the experiment?

[@elizabethowairu9296]

If you what 24 hours to heat stain what happens?

[@sunnychandra9722]

elizabeth owairu hi

[@sara-bz3cy]

the bacteria will still be alive i guess !? ..

[@12binitamajumder68]

Bacteria will be washed off during staining without heat fixation

[@VanillaSnake21]

How come when I apply the alcohol most of my specimens come out red, even something like gram positive bacillus subtilis comes out half red half blue and sometimes completely red. Professor said that could happen if I'm applying too much alcohol but I do it as instructed, until it runs clear. Could it be that my school has such old specimens that all the peptidoglycan is long gone and alcohol just washes everything away?

[@VanillaSnake21]

I can answer this now that I've finished the semester, by the end I was able to do perfect Gram stains. The reason it could sometimes comes out mixed color or the blue color washes off completely is mostly due to uneven alcohol coverage. You have to place the drop of alcohol all the way at the top of the slide and let it slide down on it's own, also don't follow the directions "until it runs clear" once you see that it's light blue it's fine to stop. It's just a matter of getting the amount and the coverage right. By the way, got an A+ in this class, due in part to videos such as these. Many thanks.

[@CutiePie-qo7zu]

Very clear explanation keep it up sir

[@rishabkumarmodi5959]

Post more helpful videos on equipments and related solutions which are used and how can they be used

[@CyrusMuema-q8q]

Am kindly asking why do you heat fix

[@sara-bz3cy]

thanks that was SO helpful 💖👏👩‍🔬

[@andreyrafaelpereiradamasce9615]

Great video although you could've shown what the samples looked like after all of those steps.

[@Rishidev2922]

very useful video.tommorw my practical xam.

[@leekspinner]

what if u accidentally swipe the bacteria from the wax pencil circle?

[@flaparoundfpv8632]

Jeez man, did you flame the loop enough?

[@faithjones2770]

Can I use blot paper to dry it or let it air dry?

[@abrokeATM]

you can do either but blotting will be quicker. if youre doing acid fast, let it air dry

[@MED.Laboratory]

Its wonderful understand the Subject after this video saved me Thanks very much

[@JoeMikeTennis]

Thank you so much for this video. ❤

[@pekhom6660]

Interesting and helpful episodes.

[@Andreahhy]

PLEASE WHAT IS THIS GRAM STAINING USED FOR I ACCIDENTALLY STUMBLED INTO IT.

[@FailureTheCrab]

Used to differentiate between two types of bacteria.

[@leekspinner]

i'm just a random person who thought that gram-negative means something about the weight thank you for education!

[@nandhinin49]

Why we are heating the steel (thin rod)

[@swapnilpradhan507]

Excellent video. Just one addition, Safranine being a weak stain should be kept for 2 mins

[@AngelStickman]

From my current work experience, one minute is sufficient.

[@khgnew763]

over staining may turn gram positive to negative

[@swapnilpradhan507]

@@khgnew763 No, its not that way, Gram positives will not take up counter stain

[@rashisahu15]

thank you for this..i hope i'll write the same in my exam👍👍

[@Blenderverse420]

where is the bacteria on the slide?

[@dr.hannibal8338]

Better than my professor explanation

[@nourassani5589]

Wow so amazing your explain 😍

[@oliviapereira364]

Thanks for explaining the technique. I don't understand the main ideia, though. Was that Gram positive or Gram negative? I've read the Gram positive retains violet and Gram negative doesn't. And that Gram - are pathogenic and gram + have more glucose. What does one thing have to do with another? How come having less glucose means increased pathogenicity? And why is it called positive/negative if the gram technique includes all stains, not the violet one? When I first saw the name I assumed the nagative would be transparent, but it is red. Can someone on the field please show me the light? thanks!!

[@digitalditz3474]

Bacteria that take up the primary stain is called gram positive and those that take up the counterstain is gram negative. Gram positive bacteria have more of the peptidoglycan layer and no outer lipid layer whereas gram negative bacteria have less of the peptidoglycan layer and more lipid layer. The basic principle is that the crystal violet iodine complex is taken up by the bacterial cell wall of gram positive bacteria and on adding the decolorizer (ethanol) it dehydrates and closes the pore ,effectively locking in the primary stain. On the other hand Gram negative bacteria, on addition of decolorizer has its lipid layer solubilized and loses the primary stain, then on addition of the counter stain, Gram negative picks it up while Gram positive already is saturated with the primary stain.

[@oliviapereira364]

@@digitalditz3474 Thank you so much for the kind and clear explanation. So the fact that Gram negative are more patogenic has nothing to do with the fact that their wall is thinner? If I add ethanol for less time than required will the gram - look like a gram +? On the same matter, if I leave ethanol for too long will the gram + look like gram -? Or once the pore is dehydrated it will be forever violet? Thank you a lot!

[@digitalditz3474]

@@oliviapereira364Gram negative is pathogenic not exactly because of their thin peptidoglycan layer but rather the presence of an outer lipid membrane structure confers greater resistance to antibiotics. Also it's more reactive meaning that in the host, the outer membrane having virulence factors like toxins and secretory molecules cause the immune response. I think if you add the ethanol for too long, the gram positive will still stay the same( might change the overall morphology and cause issues during microscopy) but if you add too less , you can expect some experimental error where some take the counter stain and some don't.

[@oliviapereira364]

@@digitalditz3474 Thank you so much! I finally get the idea

[@vegetaspride2890]

@@digitalditz3474 why we are rinsing it with water after every step ?

[@RyanBurdon-n1u]

what is used to fix the bacterial???????????????????????????????????????????????

[@godskayzee7711]

Heat from the bunsen burner or flame

[@Healthygreatmindset]

Thank you for this useful video

[@BioRadLaboratoriesCorp]

You're welcome! If there are basic biology and life-science laboratory techniques you'd like to learn more about, email the Bio-Rad Explorer education program at [e-mail]

[@alipohsem1781]

Amazing video

[@kinank2917]

Can anybody tell me what kind of iodine that used in this video?

[@fernando3061]

I would have emphasized the decolorizer is alcohol, and kills the crap out of everything so get it off within 3 seconds or less. So many students in my class didn't do that and kept getting bad results.

[@ghichy]

How come our lab manual said crystal violet stays for 30 sec?

[@irinaalexandra9050]

mine says for 1 minute

[@gayatrisawant8127]

nic and straight technique

[@AlphaShot..]

How did you get bacteria???

[@priyankaraj4789]

What is the percentage to alcohol used

[@ashutoshprasad8494]

simple and easy, thank you.😊😊😊

[@zemenuachamyeleh45]

tank you for this good presentations.i got it good experiance.

[@yuwan]

Excellent demonstration, but there is no point and even undesirable to draw a circle on the slide using a wax pen.

[@meganepelletier853]

The wax circle doesn't really interfere with the smear or staining. Although unnecessary, it is pretty commonly used and can be useful for students who are new to laboratory techniques and microscopy (eg. to divide several samples on a single slide and/or to align the field of view in the microscope). Some slides even come with premade circles or grid sections. The most likely issue would be contamination of the slide, which can be avoided using proper aseptic technique. Some students also draw small circles leading to thick smear suspensions, which can interfere with dye penetration and cellular arrangement visualisation, but that's more of an issue with smear technique than it an issue with the wax circle itself.

[@sukieyakie]

hey I wanted to see the microscopic results

[@bryanvelasco5567]

this is interesting to watch goodjob 🎉

[@mildredhart9510]

Thanks a lot, would really be helpful in my defense

[@garwex2961]

Careful ! Never wear gloves near the flame

[@anjalisharma314]

Good afternoon ma'am, Please tell 1.After culturing in test tube, at which temperature we have to keep it for preservation and upto how many days we can preserve it 2. How identification and characterization of bacteria can be done 3. If I have a single bacterial strain- and I have to culture it. Then how I know that the strain I used is the same, I want to culture. Please tell the method for it

[@MuhammadSaleem-id2ye]

Good questions. Waiting for reply.

[@DrAtifaAmbreen]

Great video 🫶🏻

[@joodalzahraa6993]

Thanks 😊 that was so helpful ☺

[@martasoria6122]

Thank you . Is very interesting

[@ANVICHAUDHARY-p9l]

Thank you so much 🙏

[@joreemmcmillan7145]

This was non-complicated and informative. However; would have liked for you to show that sample under a microscope.

[@zeenatbashir68701]

Good job 👍

[@Labelleruma]

I think its Gram negative

[@سجادمحمدجميل-ذ3ص]

Good job

[@anutamang9930]

Thanks that was helpful ☺

[@bananamcdonald4916]

I always end up incinerating the bacteria while heat fixing. Is there any way to avoid that?

[@bananamcdonald4916]

Last week on my lab exams i did the slide heat fixation perfectly. The mistake I always used to do before was that I used pass the slide through the flame for more than once which eventually incinerated the bacteria.

[@MsRicha01]

You can try methanol fixation instead of heat fixing the bacteria. works fine for us. place the air dried slide in 95% methanol for 1 mins. and then proceed for gram staining as usual.

[@melissae8480]

How can you tell when the bacteria has been incinerated??