Usually three objectives are mounted on the nose piece a. Low power objective (10x), which magnifies 10 times b. High power objective (40x or 45x), which magnifies 40 or 45 times © ilimmersion objective (10x), which magnifies 100 times Note: some microscopes also have a 5X objective v. Eye piece: consists of a system of lenses at either end of a small tube, which is. fitted onto top of the body tube. Eye piece may be of different powers like 5X, 6X, 10x. In the eye piece, a virtual magnified inverted image of the object is formed Total magnification obtained = Power of eye piece X Power of objective 10 X 10 = 100 (low power) 10 X 40 = 400 (high power) 10 X 100 = 1000 (oil immersion) IDENTIFY AND LABEL THE PARTS OF A MONOCULAR COMPOUND LIGHT MICROSCOPE acyustmumt f. < Coanse kmob mao bloc Meow slag £ nelinatior oe: aesDEFINE THE FOLI 1, FOCAL LENGTH The focal unig pon the Les Tyocussed on the 2. NUMERICAL APERTURE The numeucal diameter Of wns AAs Oo 3. POWER OF RESOLUTION The Limit of useful LOWING TERMS RELATED TO th of ent Ot gptical varie sapoatune the Lis weoustae mummericoe opi ‘atens / MICROSCOPE ; i vouctome ton us volefined! Oe ¢ vo, deme 24 ft whore panollel Mays aru do woe pow a. wns vis the Halo the eee fore Hh parcticutos, re. NA. mnagnification 4 vO. Micx0slope us Set os its Hesolving pone, 4, WORKING DISTANCE Working voliotance us the the objective lide. Tt wlucntase WW! ification. WRITE THE ADJUSTMENTS TO BE MADE IN THE MIC! DIFFERENT OBJECTIVES: LOW POWER | 10x | * To achieve focus | the NA mudt be closely, | amattehed . paducwg, the oe do the specimen) ushich om be vochieved. by vowering the vonclinsor vomel by | losing, the is | ao rragn volistamee between the objective and ith un caning, ROSCOPE FOR FOCUSSING UNDER OIL IMMERSION HIGH POWER »4Oo% ies | * FOX 0 /00x% eet a Peek Ae of oil- immersion oy | objective is valvoys ° oe ote tom +0 of ey ad a | “the conclusory ae ne eS | ° Concumaor should be eee mee | Placta vat highest a or should be ti on er fe) i | iris volia. phoma. Should be per Partially opuned. oben. |paw LABELED DIAGRAMS ‘of Rac PIPE [ >) 1 pubber L i q | | mube ae ub | | V\ a Beak Ip. mouth piec + mouth (white) S piece + stam | Ire ND Ree pipete WD a Ls Peper crioyonI TE RIMERCMEDANEUaAe DRAW A DIAGRAM OF THE CHAMBER, SHOWING ALL THE MEASUREMENTSDRAW REPRESENTATIVE 4X4 WBC SQUARE AND 4x4 SQUARE SHOWING ND 4x4 RBC sal " ] on + lmm t RY mm Let fT Hl fies || | Ea YXH WEE Suane | | 14 Wlaounumunt Beene aiaee M toournin ‘ANSWER THE FOLLOWING QUESTIONS 1. What are the differences between RBC pipette and WBC pipette? 2, What are the other uses of RBC pipette? 3. What are the other uses of WBC pipette? 4. What are the other uses of Neubauer’s counting chamber? RBC pippette WBC pipbette 4. Hod +d Beool 4. HOS white Reaol ea 9 acluatiens upto a. Has “quad dation, mork 5 make mark | 0l dd gh 3 dize of bulb is se 3 Size 4 Bulb is wwnatuy, . | B size of Aum (2 snatter Oe Se panne RBC pippeve vom be Woda pat WRC count when wee vepumt u& Da + Count shown vamel count’ platelets A wec pipette a wided to vdlilute blood with suagent com be uinid to count othen macromolecules siwilos in ize to wBe. i1 The ether wuses of Neubaturs wounding, chamber experiment No: Syuane plang iat thrnen ds sed fer WIRE Couening spp vente square fs nee counting while cbombey won bp uuseal do count mo. wf Matelits , perms éte | See tac ns DETERMINATION OF ToraL eRyruRocrTE (nec) couwT own blood The compon! 4 Sodium xulphate > Preventsfir & ws é PRECA\ 1 UTIONS Prick should be bold enough to give free flowing blood. At no stage finger should be squeezed to take out the blood, Both hemocytometer and cover slip must be dry and free of grease. Use only dry pipette. Never use a broken cover slip. Before charging the chamber, the fluid in the stem of the pipette has to be discarded, The cover slip should be placed symmetrically There should be no under — or over — charging of the chamber (count will be low In both cases). After charging the chamber, allow some time for cells to settle down but the Count should be made before the fluid in the chamber starts drying up. While counting the cells, the stage of the microscope should not be tilted 9 OBSERVATIONS. Draw representative RBC squares and enter your observations af | I2) 25] 0 231 4 4 la} || efi : fo jyols [ous Bal Selee. | 4 a4 fio |Y¥ fii] 019 la}iefo 1& 21 2] 8 14 40} 4b} 10 feat H16 ay aie nm By by 5 1} 3 lol £6 |5 6 g RBC = 5H X 10000 = 5H Mi/mm$Polerhemi 4. Tm Ree count exbodnunt, we volibute tae fluicl 40 vavoid daca itination of veULs ame cto thames Has. virtue of 4 Hunee vo vavoid errors we mimimize ie of volitucion + ty @ ce voowud clibution fatty. Aiilution factor = Final valuens. ach ieviel = Web = 20: Oniguaank volun akin Aitution pacer = Qv0 2 Nonmak Homoe of Rec um woliffount wages vamnel gem ; um swe bow > 6:8 M]mm3 Im schilowun — 34 Mimm* dm woclultt males = 4-5-6 Mims am sacle females. 9 4- 5°5 (4-3) rom3] yo 8) Bone marvore failure ©) Etegthyopoctin aepiciony. 4] Hatmolysis ¢] Intomal/ Bdomad Gee fl Leukemia 4] The al tM -O] Cgaretle smoking. ee) Ae oe fac ec) Achypratin A] Renod Ch wancinema. ey Aubmenatag frmss ‘ fl foly eythemia VO. 9) Miitute aicknors 4] sleep vapnea. |] Aetawlo ayes A ee aru the nen— nucltottol wm eda precumsor 4 blood ells that oleh um the yuel Bene ynosonr fw tu PHsc's. The normel sbieudouge poreuntage tn ae Blove!ron oF TOTAL LEUCOCYTE (WBC) COUNT acid avlutivn composition > 17 valacial ete eae an ples stoi ov aqueous mmuttigivne Blue + Aistilleal water runchin -— [¥- bunclalacrbievoeld anusc cltabructiov — Gumnbien violet stains Stains Hh uiclil of feu » Distived water att as solvimt 1 Toke WAC ain fd Tur’ ud) ina watch ass. 2._Aher peng flog, such the second drop of blood ‘pte OS mark and ut with WAC ting fid mate vars on “ho ymm® of cliliuk Blood = N 1o {important Note: WBCs are seen as regular ; ‘lean refractty around them, The cel sails on Imm of wclilute blood = * ei scat ac 6 Count the number of WBCS in ech WC sauare objet eres obienedforcunng tee f undiluted Blood = NXID x 20 = Nx AW &n lmm® oj q 3 a3400 lene ap 80 @ Acouare ini toto wise nga s s volud Pipestone Prepioleysie) LOADED i g al oto py- & Rocio tape. fathologucot counts de HEV -ATDS a. Zupus ony th Umetosus 3 Hook gkauins tymph oma | © Leuk bua rae 7 Ce clene cari which . Flee im wwmbtr of WRC Wn te Body . | dymptoms ~ Fabgue Fevers Regulon din fectiovs | WMiioze)cisollin Lymph mooltr ele. Rec. pipeve uw used for counting WRC cobls -cl Unikurnia , lange com Invuose vim wise num) vane presint pa Jakhs |m* oy millions |granulocytes: peeve 23/7 encores: 24)14 deel maede peuiph erat srt te thick becorms aa ee CL, tires with | Aves the opposite ond | at the frosteal eni gon spuvalion of B Fixation és the procers by. which unttrnal and celornol Strudlures of ctlls anel microaganisns ant pr | pra un position. ved arid | 3 Strain be frre of acetone berawuoe of Us prcsend. fb - vallohey, the cell membrane | ty Role of water in staining Pretes hs to serve 04 wcltoder ae veda wood vil fs soutinely UM for Mmmersion beau ut has wa tenatiney, to Harden camd won vase Lis dlamnage. af Mot smrard vofter use» Tis wm waportamt utest for evaluation blind sulatecl problems dich ao Hirt RRC, WEE or falatelets , 4 Neutrophils — 507--Fo7- Eosinophil I-47. ‘ Bas ophil He Uymphoesg te —9 ID —40 f+ ecteriocy te a adi 70 b no, a ear ey ripho oe » Diorneter 1a- dSum + Mametvs Is Jo-/1um 3 . . + Nucleus Is bovrge and oceuples is. i, abundant. ae Z f plow is 3 ra + eytoplaom 18 clear and Hav F v wingroumal glans 0 Vek in colous, 7 luc um appearance. . Gytoplasm Gniains granutes. aTcgunt al bes de rucleus of a type / d pt to ditect LLUULAS< 9) auneth pil in an abtiny muutrop— ~~ po. st fe toy Bor aes, compara’? eT ravenna lobin Stomalorvel Sa SAHIE'S HEMO GLOBINDMETER, A 092 mam a The HEM Lop PaPerrePeat ee of 2s ‘aucune omen caren PAETYOF YOUR ODD AT 200% SATURATION) 13 me 0; pon gra He. Ing poms ‘Hb > hBH KG F [2-06 mL. Ioortof my. Bloed haste veapaetty +0 bind to /2-06m se LOWE QUES: eee Lathe erent methods etna Remogin concentration Wye h34 2 Whatare the deadvantages of Sahi's method? 3 Which iste most acute method to estimate Hb? 4 whats the normal range of Mb concentration? 5. Which are the types of normal hemoglobin? What isthe difference in their ssrucure? DISCUSSION POINTS Structure of hemoglobin molecule Functions of Mb Different types of Mb vo b) at ©) he « 4. Nomod “ange Adult males — olult fornales — Ss. ie of nowmal eno aoe oes eBolult Hormoglobins + Harm lobin A (HbA) * Howméatobin Pa (HbA, Foetal Hatmogobin (Hb) alo a ae t's (Hb Fetal eens : 53, » Hb Gower L Hatmoglobi” Ei 7 Hb Gowtr af Hb foytland. EXPERIMENT — 2 EERE Estimation of oe - C50? Mtthod ) Nom — Chandan Nan Gunoler - fermale Advess— Dr Bnfe , Sang lors Ripert: a is 9 gm/at.OBSERVATION AND RESULT éle dome rntasured ty Luke ?s method is Qin 30 sec The normal range of BT by the Duke method is_/~ 5 Wwiini« ANSWER THE FOLLOWING QUESTIONS: 1. What is bleeding time? 2. What other methods can be used to estimate 81? 3. What conditions lead to prolonged BT 4. What aspect of hemostasis does BT indicate? DISCUSSION POINTS Hemostasis - definition, primary, definitive Steps of platelet plug formation Platelet granules and contents Platelet functions Abnormalities of platelet numbers and functions 4. Bleedu Es olefinedl vas voncoer of. Bisedia) time UiLL Ht atoppage of Blecdigh 4. Olin wethocls to ecb/nate Blecdiing tue 0) ' heal tect b> Platelet x ‘on test. e> Platdet lhe ivemness test. 3. igelengation of a is formed fowl in thrombo- topenio , throw bostrnia vamol ven -wille Brana oliscace FH occurs wen platelet count is Lins tram 60, v0 UL of — Bleocl . Thrembostuwa is the condition of Platelets are, vabromet. In vol wilebrancl d’scase , Ut alifect Is combined with factor eee er, oa’1g v4 ve pre foulons”” 4 Bleeding, He moat es +h fe a 4 a) vaspconetriho? I l > Temporoh4 plesent’s hea ao” _~ - adhesion » activation , aggregation a pis“answer THE FOLLOWING QUES Name dif i> pan bination Fy oy is absent vagal tinugn is vabser present in plocomner - is an exuption to the Hult vas ve ou is fs not peers naturally » 1» anti-A.