 Prove New Bower Chamber. So, he must cytometer by its definition. We will be able to count different types of blood cells here. Different types of blood cells. So, RBC, WBC count and your platelet can all be counted using your New Bower Chamber. Your improved New Bower Chamber. So, having said that now, okay, having said that now, when it comes to the New Bower Chamber, when it comes to the New Bower Chamber, perhaps you are wondering pano binibilang yung RBC, WBC ay wala pang machine. We do not count it on your peripheral smear because that will only be an estimate. Lahat ng lang dun ay estimate lang. But here, we will be able to somehow accurately count the RBC, WBC and even WBC, okay? So, when we are discussing hemocytometer, eto yung mga bagay na gusto kung tandaan ninyo. And hopefully, you would be able to make a list of this one para ma-make sure ninyo na you were able to cover everything about hemocytometer. The first one is the location on to where san kamis mo sa hemocytometer magbibilang, okay? Because we have a lot of squares on your hemocytometer. There are squares dedicated for RBC and dedicated for WBC. Secondly, what are the different pipets being used here? If you have red of tomapipet, if you have red of the WBC pipet, RBC pipet, we have a lot of those things and also the different diluting factor, okay? When I say diluting factor, later on, I'll explain why we have different diluting factor for RBC and for WBC, okay? So, let's start the first one. So, in hemocytometry, so this is the numerical evaluation of form elements of the blood. So, meaning to say, numerical evaluation or the counting of your form elements or your cells in your RBC, in your full blood. So, in hemocytometry, it is an estimation of the number of blood cells in a known volume of blood. Why is it that I am saying estimation pa rin? If you said a wild boxer that it is more accurate than performing it in a peripheral blood smear. We are calling it estimation because at the end of the day, the absolute count would be best taken from an automated machine, okay? from an automated machine. So, we have different methods. We have your turbidimetric method, your microscopic method and your automated method. I believe I will not be able to cover the automated method here anymore. But again, I will be recording a video about that. I will be recording a video on that. So, para ano, hindi kumansya mapalabas lahat sa exam which I think would be best, would be beneficial for you guys. At least na cover ko sa lahat, okay? At least na cover ko sa lahat. So, I will be uploading videos na lang, if ever, with the topics we will be missing. But ito lang yung gusto kong iano para walang magubulat dahil yung automation is part of the routine hematological procedures. There is a possibility that the automation, even if it is asynchronous, I would need to include that in the exam. Basta update you, tingnan ko kong sobarang dami na nang coverage. Hindi naman ako masamang tao na pipilitin ko lahat magkasha kong hindi naman nakaya, okay? So, moving forward, let's go to the microscopic method of hemocytometry which we will be discussing today because we are using your improved new-bower chamber. So, of course, you will be needing these three materials. Number one, your counting chamber or your improved new-bower chamber or your hemocytometer, your pipettes and your diluting fluid. Okay? Your pipettes and your diluting fluid. So, we have the different examples of your counting chambers according to type. We have your open type, your closed type, your addies, your exton and your petrof. I believe that in RODAC, you will no longer be seeing this but mind you, okay? Our board of exam is... Board of exam kasi natin mayroon paring manual methods. That's why we still have to discuss this. You can actually see this in an ancient book, your Barbara Brown. When I was in review, I really did buy and look for a copy of Barbara Brown and yun, book-based dapat ang pag-aaral, guys. Book-based ang pag-aaral, hopefully, you would be able to read your RODAC then para hindi siya sayang, okay? So, moving forward, okay, moving forward, we also have different chambers according to rulings, okay? When I say rulings, according to the linya, okay? According to the lines in the counting chamber. So, we have DOMA, we have the twerk, the fuchs-rosentile, the new-bower, the improved new-bower, the base-jones. So, the most commonly used counting chamber is the open-type Spencer, improved new-bower. Open-type Spencer, the new-bower, the new-bower, improved new-bower, chemo-cytometer, okay? Open-type and improved new-bower. Sir, do I really have to memorize like everything with the rulings? With the rulings and the open-types, not anymore. I just want you to remember that it is an open-type Spencer and an improved new-bower chemo-cytometer. Ayan. So, eto yung rulings na sinasabi natin. So, yung mga account dyan, one millimeter. So, madaming numero kayong imememorize, this coming examination. So, please remember these things that I am going to say to you which can be found on the notes of my PowerPoint and I am currently sending it on your chat box. So, the following, ayan. Ayan. So, these are the following. Amay nag, ano, sinabi ko, dinanilaginawa, okay? So, amay ginagawa pa rin, nang nakakita kami ng double head, double tail, go na go pa din. Ayan. Tapos, anyways, later na yung side-quento ko for that para hindi-recorded. So, with regards to ano, with regards to the, with regards to your, with regards to our chemo-cytometer, okay, this is the etura. Okay, this is the etura of your new, improved new-bower. So, this is the improved new-bower and as you can see parang hawa wala naman rulings. Wala ng rulings kasi these are like microscopic and this is the cover slip. The cover slip of your new-bower is different from the usual covers slip. And men, eto yung lagi na babasag. So, we usually have to replace it every now and then, okay? So, as you can see here, everyone, okay? So, we have a primary square. So, the primary square is a total of, kaksinabi kong primary square, eto, eto mis mong lahat ngto. The primary square is a total of, a total of nine millimeters squared, okay? Nine millimeters squared. So, each of this is actually, etong nine square na to one, two, three, four, five, six, seven, eight, nine. Yung malalaking square ha, eto, yan, eto. Waita, ay, I'll change the, I'll change, ay, ay magpachange. Etong pen na to, eto yung tinatukoy kong one, eto. One, kuha, kuha po tayo ha, sorry, tabingin. So, that is one, one, two, three, four, five, six, seven, eight, nine. A total of three millimeter, three square millimeter. So, each of this, okay? Each of this is one millimeter. One, one by one yan, one millimeter. So, one by one. So, for this one, ayan, I want to put it this way. So, sana sinusulat niyo rin na ganin to ha. This actually are for your WBC. This four are for your WBC. And remember that this four that are allotted for your WBC contains, okay? Contains 16, okay? They contain 16. One, two, three, four. One, two, three, four. They contain 16 smaller squares, okay? Eto naman center na to, okay? Etong center na to, this is for your, this center here is for your RBC, right? This one is for your RBC, okay? And this center square is actually divided into five tertiary square, okay? So, una ha, an tawag dito sa ibuong square na to, primary square. Etong primary square natin may nine na secondary square. The four outer secondary square are for the WBC. And the central secondary square is for the RBC, okay? These, okay? These central square, this central square are divided into five, okay? Five paulit, dinivide pa siya sa five. Etong five na to, nakadivide pa siya sa 16. So, nagigets niyo kong gano ka complicated yung squares natin. So, ganan lang siya, ha? So, ayan. So, this is, this is four. Wag yung kakalimutan, ha? Ang WBC 16 lang yan. 16 squares ang RBC natin 25, okay? 25, okay? 25 squares. So, tong 25 squares na yan, nadevide pa siya into 16 smaller squares, okay? So, in addition to that, please remember this, ang depth po ng ating hemocitometer is 0.1 millimeter. Again, the depth of your hemocitometer is 0.1 millimeter. 0.1 millimeter. Moving forward, saabi ko nga kanina, the central square, okay? Alam po yung central square, eto po, mga kapanalig, yan, tunasagit na yung may R. The central square is subdivided into 25 intermediate squares, okay? 25 intermediate squares. Ang measurement, the area of this intermediate square is 0.04 square millimeter. So, do we even have to memorize the zero point keme keme? Not this, etong measurement na po, not anymore, ha? Pero gusto ko lang, alam niyo kung paksinabi kung central square, outer square, secondary square, nakakuha. So, yun yung aaralin natin, ha? On the notes that I will be giving you, maganda yung notes na yun. Nakalagay dun kung saan niyo bibilangin. So, W may nakalagay ng W, okay? So, each intermediate square is further subdivided into 16 squares. So, imagine that, nabah? So, the red blood cells are usually counted on the central and the four, in the central and the four corner intermediate square. Four corner intermediate square, ayun. Four corner intermediate square. So, ayun, okay? So, that is for your red blood cells, for your four corner large squares on the other hand, that is one square millimeter, bakit one millimeter? Kasi ayun, one, one, one by one, that is one square millimeter. So, that is for your WBC count, okay? And it is only subdivided into 16 smaller squares, okay? So, hopefully, okay pa tayo, mga kapanalig. So, ano yung import yung nilagay ko sa inyong chat box that are the measurements that are important. For you to remember, okay? Moving on, ayun, kita niyo, ganyan yung ityura niya. So, in the new bower, actually, when you look at the new bower, you can see the grid lines macroscopically, but very, very mind niyo. Pero, pagtingin mo sa ano, okay, winner, ganyan ityura niya, okay? So, um, there were cases na ganyan ito yung nangyari. There were cases na, sir, paano pag overlap? So, when I count it, pag nandito yung, yung ano, cell, yeah, you still have to count it. Pumbagan ang dung siya sa linya, ang dung siya sa linya, it's still included in the count, ha? So, for the pipet, ayun, meron tayong dalawang, two types of pipet, okay? So, kung meron kang, masamang panaginip sa pipet, okay? Sir, ba't iba na naman po yung spelling? Pipets na ganyan, or TTE, any spelling would do. Automatic pipet, we have your trainer and your UNOPET. Okay, UNOPET. So, itong UNOPET na tolumala. Alam niyo ba yung UNOPET na rinig ko lang nung nag-review na ako for board? So, UNOPET, automatic pipet yan, for your hemocitometer. We also have the non-automatic pipet, this one, the TOMA pipet. This is the reason why, siguro, yung generation natin right now, would wonder, baka tiyan nila sinasabi na no-mouth pipeting? Kasi nung unang parang unang nag-mummouth pipeting sila. So, imagine nung nag-mummouth pipet sila using your TOMA pipet, your TOMA pipet can also be used in your seminalysis. So, imagine mo, di ba? Nag-tomapipet ka, mouth pipeting ng seminalysis, tapos na todo-todo mo yung hicop. Ay, just call it, okay? So, huwag nana nang pag-usapan nyo. So, TOMA pipet, ayyan, TOMA pipet, we have your RBC TOMA pipet and your WBC TOMA pipet. Obviously, yung isa para sa RBC, yung pangalawa para sa WBC. Are we clear? So, TOMA pipet and your, TOMA pipet and your, yeah, RBC TOMA pipet and WBC TOMA pipet. So, nang pinakaiba, later, pag nandunatay sa deluting factors, ma lalaman nito, okay? Now, let's go to the deluting factor, deluting fluids. So, the deluting fluids are used to disperse blood cells to facilitate counting of RVs, counting of cells. So, anong ibig sabihin nito, sir? Kelangan natin ng deluting fluid para hindi dikit-dikit yung ating RV, yung mga ating cells, so that they would be dispersed with one another, okay? So, we have RBC deluting fluid and we also have WBC deluting fluid. Sir, remember this, everyone have. Pag RBC deluting fluid, isotonic yung solution. Sir, bakit po isotonic so that the RBC would retain its shape, okay? Para yung shape ng RBC mo all throughout the counting is still the same. On the other hand, WBC deluting fluids. Sir, ma'am, kailangan po hypotonic yung inyong solution. Sir, bakit po kailangan hypotonic and I need you to write this down, okay? I need you to write this down. Sir, bakit po kailangan hypotonic pag WBC deluting fluid? This is the reason why. Hypotonic to allow the RBC to lyse. Para masira lahat ng, para maglyse lahat ng RBC. At dahil naglyse lahat ng RBC, ang may iiwan na lang sa solution mo are your WBC. Mas maglalis, mas madali. Kailangan yun, makakapagbilang sa New Bauer Chamber. Nakakukuha po ako. Can I see a raise of hands if I'm clear with the difference between the RBC and the WBC pipet? I delusion fluid, okay? Thank you so much. So yung yung reason ha, kailangan mawala ni WBC. Para hindi siya pang gulo. Nakakukuha. Ganon din yung importance kung bakit may delusion tayo, okay? Yung delusion ng RBC papasok siya sa delusion factor. Kasi magtataka kayo. Sir, isotonic yung RBC. Mag-remain po yung ating, may iiwan po yung ating WBC. Not anymore because it will now fall on the delusion factor, okay? So we also have platelet deluting fluid. So the reason, the number one consideration for platelet deluting fluid is that they should preserve the integrity of your platelet while inhibiting their aggregation, okay? Prevent po yung aggregation ng platelets mo habang nabibilang mo siya, okay? And remember this, your formal citrate or your dasis fluid is the best RBC deluting fluid. Dasis fluid, okay? Dasis fluid for your RBC deluting fluid, okay? Moving forward for the different RBC deluting fluid. So in general, madami man tayong RBC deluting fluid. They are the same in a way na all of them are isotonics, okay? Isotonic solution. So we have your HAMs, the RBC deluting fluid. We have your goers. We have your toysons. And we have your toysons deluting fluid. So nandun sa kabilang slide yung iba. So your HAMs deluting fluid, it initiates mold formation and rollu formation which is a no-no because you want each and every RBC to be individual. Your goers on the other hand, prevents rollu formation but the problem is that it precipitates protein in cases of hyperglobulinemia and hemoglobinemia. So another fall for it. So your toysons initiates mold formation so it should be filtered. It has high specific gravity and it has your stain, okay? It has stain. So for starters or beginner, it would be very useful, okay? It would be very useful to use your toysons. Nakalimutan ko yata sabihin sa inyo. Now when counting blood or counting cells in your new power chamber, wala kang stain usually. Wala kang stain na ginagamit as in yung ano lang, deluting fluid lang. So ang anon, when you look at in the microscope, alam moong parang transparent lang silang mga bilog-bilog. Sir, pano mo na lalaman hindi yung double, hindi double ubc yung nababasa mo. Kasi yung wbc naman, makikita mo yung nucleus niya kahit transparent, right? So medyo mahirap siya sabihin ngayon kasi we're not looking at it in the microscope, but trust me when I say madali lang siya ma-identify pagdating sa practice, okay? Madapapraxis tayo hopefully next year. So ayun ha, guys, I want you to remember toysons because it's the only RBC deluting fluid with stain, right? Okay, clear? So moving forward, ayun, ano yung stain na gamit na yung methyl violet? Ayun, your methyl violet. Moving forward, that is toysons, toysons, toysons. So moving forward, we have here your bethels, we have your formal citrate which is your basey fluid, the best RBC deluting fluid because it preserves the action and has a preservative action without mold formation, okay? And it preserves cell morphology as well. Aside from that, we can also use NSS in case of excessive ROLU formation or autoagglutination. And you can also use 30.8% sodium citrate. 3.8%, 3.8% sodium citrate. Okay? So formal daysies, again, guys, remember, all of this collectively are all isotonic solutions. We go now to the WBC deluting fluids. In the WBC deluting fluids, ang goal natin is to hemolize the red cell except for metarugricite. What are metarugricite? These are nucleated RBC, okay? These are nucleated RBC. The last stage in your erythrocyte maturation that has in the cruise is your metarugricite. So the leuens, okay? The leuens for counting only leucocytes should be capable of lysing erythrocytes without destroying your leucocytes. One of the following leuens may use, we can use your glacial acetic acid which is actually one of the most common. We can also use your hydrochloric acid your turx solution. Maganda yung turx solution. Hindi ito yung shawarma kung nagugutom ka ngayon kung it enhances the leucocyte nuclear definition. Okay? Leucocyte nuclear definition. Am I clear with that, guys? So ayan yung ginagawa, ginagawa natin for your WBC. The most common is this one, glacial acetic acid. It is the most commonly available kasi in Dalak. So moving on, we also have here important reminders, by the way, in manual counting, do not overcharging when lethal to decrease in cell count. What do I mean by overcharging? Okay? Puno a rin yung pipet mo magdideliver ka na ng fluid sa counting chamber. Usually kasi, meron ka ng counting chamber, tapos yung slide. And then dun sa pagita nila, dun mo ipapipet yung fluid. Okay? So avoid, okay? Avoid overcharging kasi magpaflod yun, mawawala sila dun sa counting chamber. Aside from that, it is important to allow the counting chamber to stand for at least three minutes after charging. So this is Poison's Law of Distribution. Bakit? Kasi, I don't know if you were able to look into the microscope na of a urine, pero gumagalaw yun, kasi nga water siya di ba? Solution. So kapag pinilit mo magbasa, right after charging, ang mga yari niyan, gumagalaw yung mga cells. So malilito ka. Pwede yung maulit mo mga count or hindi mo mga count yung isang cell dahil nga gumagalaw sila dun sa fluid. Okay? So yun yung ano natin, ha? Yun yung important na reminder. Aside from that, the allowable difference between two chamber counts are 15 to 16 for RBC and 10 to 12 for your WBC. Sir, bakit po two chamber counts? Anong ibig sabihin mo two chamber counts, sir? Babalikan ko to, no? Alam nyo, itong nakikita nyo yung ito, guys. Ay nasa dalawang position. Okay? Isa sa taas, isa sa baba. So yung would count on top and on the bottom. Okay? On top and on the bottom as well. So clear tayo dun. So remember that, ha? Dapat no overcharging and wait for three minutes before you read. Okay? So most of the questions here would actually be situational. So I would really encourage you to, siguro, when you study the lessons, aside from reading the PowerPoint, aside from reading the notes, I'll be sending you and reading the book. You also listen again to the recording if necessary. Okay? So characteristic of an ideal, diluting fluid except, ayan, nakikita nyo nga yung mga tanungan natin. So isotonic should be for RBC, hypotonic for WBC, even economical, easy to secure and prepare with preservative action, high specific gravity. It should be stable with buffering action without, with buffering action rather, non-allergenic and non-corrosive. Okay? So let's start with our, let's start with our, let's start with our hemocitometer, the different steps. Okay? The different steps for your hemocitometer. So yung nan dito kasi is for platelets na, which is okay for me to miss, because we have an exam pa for hematool. Okay? So I'm just actually going to finish this, guys, until, until the formula for your WBC, yun na lang. Ha? Para hindi na tayo magkandalok-aloka pa. So, for the steps, ayan. So for, you have to suck the blood, you have to aspirate the blood into the 0.5 mark of your pipet. Okay? 0.5 mark of your pipet. And then afterwards, you would also aspirate the diluting fluid, 100 at 101 for your RBC pipet and 11 for your WBC pipet. Okay? For your WBC pipet. And then afterwards, you need to shake the pipet well. Sir, um, do I need to do it manually? No? Because in, in TOA, meron tayong mga pipet shaker. Okay? Kung ginagawa nila mag-shake lang, okay? May pipet shaker tayo sa, sa school. Aside from that, um, before charging, you have to discard a few drops first from the stem para talagang sure ka na walang bubbles na doon. And then afterwards, you need to charge the counting chamber. When we say charge, we're not talking about electrical charging here, but charging using your, charging using your charge lang. Arang, um, anong taong mo doon? It's a charge using your pipet. Okay? Using your pipet. So again, um, in your, in your RBC, you count your RBC in your 5 intermediate square, central and the central square and the 5. Large squares. So yung W, yung sa WBC, mag-account ka dun ng WBC. Mag-account ka din doon ng RBC. Okay? So, um, for the WBC, outer squares lang. Okay? Outer squares. So, ito siya. This is how you charge it. And dito na yung new power, and dito na rin yung cover slip, and then you charge your, um, new power chamber using your pipet. Okay? So for the formula, okay? The formula for your RBC count is this one. RBC counted times 10, times 10, times 200, times 5. Ano po yung 10? That is the depth correction factor. The 20 is the delusion factor. Okay? Although not constant, it depends upon the blood that you are using. But the usual, yung .5 na sinasabi natin na mark, the .5 mark for the blood is already using the 200 delusion factor. Right? So delusion factor. Same thing, okay? So, for the delusion factor, eto siya, volume of the valve, volume of the valve is 100 for RBC, and 10 only for your, um, 10 only for your, um, tau dito. 10 only for your WBC. So blood volume use may differ. So, 100 tapos nagpalit ka. So, um, mag-vari yung, mag-vari yung delusion factor. Okay? Mag-vari yung delusion factor natin. So again, remember that the 10 here is the depth factor. Sure, kailangan i-memorize, yes, kung ano yung ibig sabihin. This is the deluding factor, and this is the area correction factor. Okay? This is area correction factor, constant unless otherwise specified. Okay? So, ayan, for your area correction, kung ayaw mo talaga, gusto mo mag-bilang. So, sige, mag-bilang ka. So total number of internal, uh, intermediate square for your RBC over the number of intermediate square used for counting. So meaning to say, kasi di ba meron tayong inter yung counting natin, you counted in, um, at total of, sabihin mo nang ilang intermediate square, yung 25 for the 16, 16, 16, 16 times 4 plus 25, din yung number ng intermediate square mo. So, ibig yung sabihin dito, kung saan ka lang nag-count? Okay, kung saan ka lang nag-count, yun yung ano hin mo. Okay? So, let's try this one. Everybody, okay, copy the formula. So RBC times 10 times 20 times 5, and this is the, this is the, this is an example of a problem that you will be seeing in your, in your exams. So, quadrant 1 meron tayong 109. So what you're going to do is actually to sum it all up. Okay? Sum it all up and then start multiplying it with the constants times 10 for the depth correction factor, another 10 for the delusion factor, and another 5 for your, um, area correction factor. Nakukuka po ako. So ilan ng sagot. Anong sagot ninyo dito? Anyone? So I'll also try to compute it. Ilan po ang ating sagot, mga kapanalig sa paniniwala? 109 plus 93 plus 105, 116, 123. Okay, time 10 times 200 times 5. What is the answer? Ano po ang sagot? Alatulog na yata sila. The answer is 5,460,000 um, millimeter, cubic millimeter. Rather, cubic millimeter. There. Cubic millimeter or you can express it um, times 10 to times 10 to 12 power per liter. Okay? So gamitin na lang natin para hindi kayo malito para mabilis lang din itype sa TLC millimeter mercury. MM3, okay? MM3. Everybody got the answer. Nakukah. So ito yung sinasabi ko, Jericho and the rest. So when you count, na niyan. You count there, then baba, then you go left, then down, yun yan. This is for the new power para sure ka na naka-count mo lahat. Okay? So Sir, bibigay mo ba kami ng ganyan? Ang sama ako ng mga taon kung ganito yung exam niyo, kus bibilangin yung isa-isayan. One, two, tapos 45 minutes lang. So ito yung example na ito, hindi niyo ito makikita, guys. So okay yung magalala. Okay? Numero na po, yag adang i-bibigay. So for the WBC, okay? Last, last, okay? Last slide for today, for Hima. So for me lang for WBC count, ayan. Almost the same. We have your WBC count on the four intermediate square times ten, the correction 20 is the dilution factor all over four, which is the correction, area correction factor. Okay? Unlike in your RBC, we multiply it, here in WBC, we divide it. Okay? We divide it. And that is it actually for today, for hematology. Okay? So hopefully everybody's still awake and thank you so much once again for listening. So if you have any questions and clarifications, you can start sending in your questions on our chat box so that I'll be able to answer them all. So the time is 1028.