Total Leukocyte Count | TLC Blood Test Procedure | Total WBC Count Experiment

This video explains the procedure of Total Leukocyte Count ( #tlctest or TLC Blood Test ) using Neubauer Chamber or Haemocytometer with calculations. It is also known as Total WBC Count Experiment, White Blood Cell Total Count, TLC lab test, TLC DLC blood test, etc. In Hindi सफेद रक्त कोशिका जांच,
tlc test kaise karte hai?
For detailed information visit: www.ebiologylab.com/experimen...
Principle of TLC Blood Test:
The total number of leukocytes WBC per mm3 of blood is determined by diluting the known volume of blood 20 times with fluid, which prevents hemolysis and coagulation. The blood cells are then counted in the volume of diluted blood in Neubauer’s counting chamber or Haemocytometer. The number of WBCS per mm3 of the undiluted blood is then determined by this method.
Apparatus required for #TLCBloodTest:
1.) Neubauer Blood Counting Chamber: amzn.to/2S9VpR2
2) WBC Pipette: amzn.to/3xutNoD
3) WBC diluting fluid or Turk’s fluid: amzn.to/3q4yJ0W
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
Neubauer’s hemocytometer:
The instrument is consists of a special glass slide. The counting chamber is a very heavy thick glass slide usually at the center with three platforms, which is separated by wide grooves. The central platform is 0.1 mm lower than its neighbor. On the surface of the middle platform, there are two sets of ruled areas. The area has 9 large squares, each has 1 square millimeter area and is elaborately ruled. The four corner squares are divided into 16 equal-sized squares. The central square is ruled into 25 groups of 16 small squares, each group is separated by triple lines. The ruled surface area is 1/10 millimeter below the inner surface of the cover glass placed over the middle platform. Thus each smallest square of the corner has a volume of 1/4 x 1/4 x 1/10 or 1/160 mm3. Whereas the smallest square in the center has a volume of 1/20 x 1/20 x 1/10 = 1/4000 mm3. This central segment is used for RBC counting. In four big squares, leukocytes or WBCs is are counted which are placed at the four corners of the erythrocyte counting Chambers.
WBC diluting fluid or Turk’s fluid: amzn.to/3q4yJ0W
The fluid consists of Glacial Acetic Acid, 1.5 ml (for destruction and dissolution of RBCs and to fix WBCs); Gentian violet solution, 1.5 ml (1% solution, stain WBCs); these components are mixed and diluted 100 ml by distilled water.
The procedure of TLC Blood Test:
1. The counting chamber is placed on the microscope stage. The illumination is adjusted and the right upper groups of 16 squares for WBCs are focused. One can see all the squares in the alone field.
2. 1 ml of Turk’s fluid is taken in a watch glass.
3. Collection of blood:
The tip of the finger is sterilized with 90% alcohol and allowed to dry. The finger is pr.cked with a needle to produce a drop of blood of adequate size 3 to 4 millimeters in diameter. The blood is sucked up to the 0.5 mark into the WBC pipette.
4. The blood is diluted with Turk's fluid up to 11 mark by sucking the diluting fluid by the pipette kept in a watch glass. When the blood is taken up to the mark of 0.5 the blood is diluted only 20 times. The contents inside the bulb of the pipette is mixed for 3 to 4 minutes.
5. The first two drops of diluted blood from the Pipette are discarded and then this mixture is run on the hemocytometer slide on both the chambers on both sides under a special coverslip.
6. The corpuscles are allowed to settle down for 3 to 4 minutes and then the number of WBCs are counted under the low power of the microscope in the four counting areas.
7. The white cells are recognized by the refractile appearance and by the slight color given to them by the stain contained in the diluting fluid.
8. The cells are counted under high-power lens. One may count the WBCs in 16 squares under low power and then under high power for comparing the results. The cells in four groups of 16 squares each i.e., a total of 64 squares.
Counting procedure: Cells which are touching the left-hand lines or upper lines of the square are included in the count, while cells touching the lower and right margins are excluded.
Haemocytometer Calculation:
The WBCs counted in 16 squares the volume of one square being 1/64 mm3
Volume of 16 square = 1/160 x 64 = 4/10 mm3
Thus, the total volume of diluted blood in which WBCs were counted = 4 / 10 mm3
Let the count in 4/10 mm3 = ‘X’ WBCs.
Then 1 mm3 of diluted blood will contain = ‘X’ x 10/4 white cells of WBCs.
As dilution is 20 times, 1 mm3 of undiluted blood will contain = ‘X’ x 10/4 x 20 = ‘X’ x 200/4 = ‘X’ x 50 WBCs. In general, 1 mm3 of undiluted blood will contain = ‘X’ x 10/4 x dilution factor of WBCs where, dilution factor maybe 20 or 10.

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[@DKBiosolutions]

Thank you Ismail, thank you for appreciation 😊👍...

[@bilalabbasbutt4976]

Best

[@mercyjames9681]

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[@DKBiosolutions]

Thank you for your appreciation 😊👍... I am glad to hear that it helped you... Best wishes for your future studies 😊👍...

[@alitaarhun3]

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[@DKBiosolutions]

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[@bilalabbasbutt4976]

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[@DKBiosolutions]

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[@varshahanumanahalli9022]

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[@DKBiosolutions]

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[@DKBiosolutions]

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[@devonfernando727]

A best video about WBC counting 🔥

[@DKBiosolutions]

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[@LearnTeachEnjoy]

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[@rdnnews6264]

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[@harshim9336]

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[@DKBiosolutions]

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[@Mental-medicos]

What about 79 multiply 50 from where this 50 comes????

[@DKBiosolutions]

Please go through video description to know more... 👍

[@justinparale900]

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[@adrija_banerjee]

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[@DKBiosolutions]

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[@shrikantmali8321]

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[@dakeshnishaddakeshnishad4044]

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[@amirnag724]

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[@DKBiosolutions]

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[@amandeep-gi3ol]

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[@DKBiosolutions]

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[@heenasultana6531]

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[@DKBiosolutions]

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[@moreshwarmakade726]

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[@DKBiosolutions]

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[@shaikfarah8061]

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[@DKBiosolutions]

I'm glad to hear that it helped you. Beat luck for your examination 😊👍

[@etczone9985]

Understood it was very easy to understand it with this video

[@DKBiosolutions]

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[@vipulppipaliya2021]

Great explanation !!

[@DKBiosolutions]

Thank you 😊👍...

[@vellore0014]

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[@DKBiosolutions]

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[@karthishaulloju2301]

Thank youu saarr....🥰

[@elmialonte8267]

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[@DKBiosolutions]

Thank you 😊👍

[@priyalichopade7422]

Thanks sir for this video this is very helpful video for counting WBC

[@DKBiosolutions]

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[@anjanaanju4292]

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[@DKBiosolutions]

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[@harshadadhaybar]

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[@DKBiosolutions]

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[@simrankaur2471]

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[@savitapatel2328]

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[@DKBiosolutions]

My pleasure 😊👍...

[@khulnajila471]

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[@walidelsayed3282]

Wonderful!
This will be very helpful in the practical exam of physiology
Thanks 👍

[@DKBiosolutions]

Thank you so much for appreciation 😊👍

[@tharindusapumal6196]

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[@DKBiosolutions]

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[@narmeenfatima2980]

Kamal explanation 🌟🍁🙆🍃

[@DKBiosolutions]

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[@srujan4942]

This is very helpful sir.

[@DKBiosolutions]

Glad to hear that it helped you 😊👍...

[@MedicalTravelBite]

very good demonstration

[@DKBiosolutions]

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[@ramaraja8878]

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[@romeojuilet7229]

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[@DKBiosolutions]

Thank you 😊👍, Best luck for your Practicals....

[@samsuddinansari6083]

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[@thestingdrarslaan]

Watching it 35 minutes before exam🤩🤩

[@debankadebnath4422]

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[@user-lw9jy1rq3l]

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[@skgofficialpk7136]

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Life saver, one day before the exam...

[@DKBiosolutions]

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[@dradil8937]

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[@DKBiosolutions]

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[@wasimsayyad3489]

Its really easy to understand sir
thanks DK biosolution

[@rohitnaikwade687]

Thanks sir...please make video on RBC count please

[@DKBiosolutions]

Welcome Rohit...
Next video will be on RBC count 😊👍

[@xasanhiil4812]

This method Is the best way to undrstand count of wbc

[@DKBiosolutions]

Thank you Xasan .... 😊👍

[@PardeepSingh-ym9bp]

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[@DKBiosolutions]

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[@DKBiosolutions]

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[@debankadebnath4422]

Should we consider the upper Nd right margin for counting wbc s?

[@DKBiosolutions]

Why not... Any two adjacent margins are fine to consider...

[@noorhamed9630]

State four source of error in WBCs and indicate if the cell would be falsely increased or decreased..

[@user-ej1dj3zj4n]

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[@tcreation8819]

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[@DKBiosolutions]

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[@neelpatel4609]

Thank you 😊 so much, explanation is very good.but I've one query that in the last step of calculation ..how comes 50 ??

[@DKBiosolutions]

Thank you for your appreciation... Please go through video description to know more.... 😊👍

[@neelpatel4609]

@@DKBiosolutions ok thank you 😊 so much sir.

[@DKBiosolutions]

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[@fehmeedashah5997]

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[@priscillatete3398]

Thnx for easy explanation

[@balajilandge569]

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Thank you 😊👍

[@abhijitmaji4805]

Nice👍👌

[@DKBiosolutions]

Thank you 😊👍...

[@reconditewave1604]

Sir, please mention weather it is refer by coffins method or not?

[@DKBiosolutions]

No it is not....

[@noorhamed9630]

Why must the diluted blood sit in the tube for 10 minutes before it is plated on the hemacytometer??

[@DKBiosolutions]

In that time RBCs i.e. erythrocytes gets hemolysed. In simple words, just to destroy RBCs...

[@pmadhukumar9224]

Supper video

[@DKBiosolutions]

Thank you P Madhukumar... 😊👍

[@strokesandsutures]

I have a question. Why have you only selectively counted Wbcs? Like there were Wbcs present on the other sides also, but in a particular grid, you've just focused on 3/4 and then moved to the next column! Was it for easier calculation purposes?

[@DKBiosolutions]

Yes... It was for calculation. We convert those counted cells number into per ml of blood cells number...

[@strokesandsutures]

@@DKBiosolutions ooh okay got it! thankyou so much for this video - truly v helpful!

[@DKBiosolutions]

My pleasure 😊👍

[@jaivinay6001]

Good sir

[@DKBiosolutions]

Thank you 😊

[@divyabharathiak422]

Why that central square is used for rbc and corner squares are for wbc?is there any reason?

[@DKBiosolutions]

The hemocytometer is composed of nine equally sized bigger squares. The central one is different from the other ones because it is divided into 25 smaller squares, while the ones in the corners are divided into 16 smaller squares. The rest of squares are not used. In addition, the smaller squares inside the central square are subdivided into 16 even smaller squares each. This allows to count very tiny cells with the same precision level as larger ones (but with a higher magnification).
Cells that are 10 μm or more in diameter should be counted in these corner squares, while, cells that are 10 μm or smaller should be counted in the central square...

[@divyabharathiak422]

@@DKBiosolutions thank you so much for your immediate reply sir

[@DKBiosolutions]

No problem 👍... Its my pleasure 😊

[@shakir1325]

❤❤❤❤

[@coraline-2000]

Thank u very helpful but I've question what about the L shape technique for the counting ?

[@DKBiosolutions]

Thank you... I don't know anything about L shape techniques... If you know anything please share with us... 😊👍

[@coraline-2000]

@@DKBiosolutions it's a technique to avoid double counting of cells... well basically you draw an L around the sheet and you count all the cell on the top and in the left and avoid all the cell in the bottom and in the right and then u draw a zig zag and count all cell

[@DKBiosolutions]

Ok thank you for the information

[@coraline-2000]

@@DKBiosolutions your welcome 😁👍

[@ashandeepkaur4527]

everything was very nicely explained...........just the formula applied is a bit wrong...............
TLC = [ Counted particles x dilution factor ] / [ surface area (mm^2) x chamber depth ]

[@DKBiosolutions]

Thank you for your suggestion... Will check it... 😊👍

[@balajilandge569]

Sir platelets ka bi expiliment Kare na plz 🙏🏻🙏🏻

[@DKBiosolutions]

Sure, next video will be on 'Counting of RBCs and PLATELETS' 👍

[@lazypotato9377]

There are so many dots, how do we prevent ourselves from counting those too

[@DKBiosolutions]

If you get confused while counting cells, focus that dot under high power and then confirm wheather that dot is cell or just a partical or debris...

[@moreshwarmakade726]

Why first two drop in pipette is discarded

[@DKBiosolutions]

Hi...
Here we need to discard first two to three drops because we have diluted blood in bulb of WBC pipette and stem contains diluting fluid only. If we take those first two to three drops for counting WBCs, bias to our count will occur. To remove the error we have to do the same....
I hope the above information satisfies your concerns regarding this matter. Please feel free to ask me, if you have additional questions.
Best luck 😊👍💐

[@LearnWithSigma]

N×50 .. where did you get the 50? Please assist

[@DKBiosolutions]

Please go through video description to understand it...

[@harpalkaur1988]

Vry nc

[@DKBiosolutions]

Thank you 😊👍

[@priyalatha5817]

👍🏾👍🏾👍🏾👍🏾👍🏾

[@cumali6187]

Wow ..I didnt know that WBC pipet still exists...and they still use the tubing...wow..Im scared to use that...imagine if the patient has contagious disease like CJD...patay kang bata ka..

[@mulatalemu3610]

Rbc manual count

[@momori5785]

where did you get the 50 from?

[@DKBiosolutions]

Please go through video description....

[@thesunilbutiya]

Nice video sir

[@Prince_sen_0001]

Sir RBC pr vidio nhi h kya

[@DKBiosolutions]

th-cam.com/video/oPPAkQPur_k/w-d-xo.html hai na...

[@varunganipitla1199]

I want AEC slide microscopic vedio

[@DKBiosolutions]

We have lockdown here... Colleges are closed... I will try to do it once it ends...

[@younuszohaibilahikhan8819]

Thanks alot sir...
Smjh nhi aara aap ka shukriya kin alfaz se ada kru...

[@DKBiosolutions]

Hello Dr. Younus Zohaib Ilahi Khan...
Aapka Shukriya, muze bahot accha laga ki apko content pasand aaya😊. Aapka appreciation meri himmat badhata hai. Thank you so much, 😊👍...

[@younuszohaibilahikhan8819]

@@DKBiosolutions ji sir aap se yahi guzarish hai ki aap aise usefull content waale videos baante rahiyye

[@rajkumaripatrey3091]

Turk's solution hi kyo use karte h sir

[@DKBiosolutions]

Because that turk scientist has developed a formula to destroy the RBCs. It was named after him.

[@rajkumaripatrey3091]

DK Biosolutions ok thank you sir

[@BVNJayakiruthikaM]

Can you pls kindly upload the counting rules

[@DKBiosolutions]

Cells which are touching the left-hand lines or upper lines of the square are included in the count, while cells touching the lower and right margins are excluded.

[@sabrunjamil8505]

You said that 79 × 50, how you got 50 in formulas?

[@DKBiosolutions]

Please go through video description... 😊👍

[@akshitkesarwani4975]

please tell TLC 7220 IS OKAY OR NOT ?

[@DKBiosolutions]

The normal WBCs in the blood are 4,500 to 11,000 WBCs per microliter... Go through video description for more understanding.. 😊👍

[@lordcanning4362]

Uncountable thanks

[@DKBiosolutions]

Always welcome 😊👍...

[@vinayakbhandarkar4124]

50 kese liya bahi formulae mai.50 kyu consider kiya please tell mi

[@DKBiosolutions]

Please go through video description... Still if you have any questions please feel free to ask.... 😊👍

[@premroy8312]

Why we are multiplying by 50 ?

[@DKBiosolutions]

Please go through the video description to understand the calculations... 😊👍

[@IbcSahilChaudhary]

Thank u sir, but agr ap hindi me video banate to students ko kafi help milti. 90% you tube BMLT/DMLT channel hindi hote h.

[@DKBiosolutions]

Thank you Sahil.
Mai yahi topics pe naye videos shoot kar ke ya innhi ko hindi voice deke upload karunga. Thank you for your suggestion 😊.

[@cumali6187]

Dont tell me that youre still going to use that tubing after touching it with different surfaces...why cant you just remove it after sucking the wbc diluting fluid...and the fluid should be up to the marker only..you passed that marker by the way..

[@DKBiosolutions]

Thank you for suggesting... I will take care of that... It just a one time demonstration...

[@shreyahazra6960]

Pls provide trbc exp. Also 😭

[@DKBiosolutions]

Please visit channel... Total RBC experiment is also there...

[@Love-of-learning]

Peeeeerfect

[@DKBiosolutions]

Thank you 😊

[@josephaneesh9795]

tlc count is 19500 problem. ..?

[@DKBiosolutions]

Hi...
Normal range for TLC is 4k to 11k... 19500 is not normal... Repeat the experiment... Or visit your Dr...

[@zaminmushtaq2262]

4 hours before exam😅

[@DKBiosolutions]

Best wishes 👍🙂

[@skgofficialpk7136]

When you'll expert then It'll be easy and quick