This document provides instructions for making and examining a blood smear. It describes three types of blood smears - the cover glass smear, wedge smear, and spun smear. The wedge smear technique is explained in detail, including steps for placing a blood drop and spreading it across the slide at a 30-40 degree angle while controlling thickness. Characteristics of a good smear and common causes of a poor smear are also outlined. The document also covers fixing slides, staining techniques including Leishman's stain, and manually performing a differential white blood cell count and examining red blood cell morphology.

1. BLOOD SMEAR
EXAMINATION
Making Blood smear

2. I- Preparation of blood smear
 There aarree tthhrreeee ttyyppeess ooff bblloooodd
ssmmeeaarrss::
11.. TThhee ccoovveerr ggllaassss ssmmeeaarr..
22.. TThhee wweeddggee ssmmeeaarr ..
33.. TThhee ssppuunn ssmmeeaarr..
 TThhee aarree ttwwoo aaddddiittiioonnaall ttyyppeess ooff bblloooodd
ssmmeeaarr uusseedd ffoorr ssppeecciiffiicc ppuurrppoosseess
11.. BBuuffffyy ccooaatt ssmmeeaarr ffoorr WWBBCCss << 11..00××110099//LL
22.. TThhiicckk bblloooodd ssmmeeaarrss ffoorr bblloooodd ppaarraassiitteess ..

3. WEDGE BLOOD SMEAR
 SSppeecciimmeenn :: EEDDTTAA bblloooodd wwiitthhiinn 22 ttoo 33
hhoouurrss && ccoolllleecctteedd ttoo tthhee mmaarrkk oonn ttuubbee..
 NNoott''ss :: MMaayy cchhaannggee RRBBCCss mmoorrpphhoollooggyy
ssuucchh aass SSppiiccuullaatteedd ((ccrreennaatteedd)) cceellllss iiff ::
11.. EExxcceessssiivvee aammoouunntt ooff aannttiiccooaagguullaanntt ttoo
ssppeecciimmeenn
22.. OOlldd bblloooodd -- lloonngg ssttaannddiinngg..
33.. WWaarrmm eennvviirroonnmmeenntt ((rroooomm tteemmppeerraattuurree))
mmaayy hhaasstteenn cchhaannggeess..

4. PPRROOCCEEDDUURREE
 ppllaacciinngg aa ddrroopp ooff bblloooodd ffrroomm mmiixxeedd
ssaammppllee oonn aa cclleeaann ggllaassss sslliiddee..
 SSpprreeaaddeerr sslliiddee uussiinngg aannootthheerr cclleeaann ggllaassss
sslliiddee aatt 3300--4400 ddeeggrreeee aannggllee..
 CCoonnttrrooll tthhiicckknneessss ooff tthhee ssmmeeaarr bbyy
cchhaannggiinngg tthhee aannggllee ooff sspprreeaaddeerr sslliiddee
 AAllllooww tthhee bblloooodd ffiillmm ttoo aaiirr--ddrryy ccoommpplleetteellyy
bbeeffoorree ssttaaiinniinngg.. ((DDoo nnoott bbllooww ttoo ddrryy.. TThhee
mmooiissttuurree ffrroomm yyoouurr bbrreeaatthh wwiillll ccaauussee
RRBBCC aarrttiiffaacctt..))

5. STEPS FOR BLOOD FILM

7. The thickness of the spread
Notes:
1. If the hematocrit is increased, the angle of
the s preader slide should be decreased.
2. If the hematocrit is decreased, the angle of
the spreader slide should be increased.

8. large angle
low HCT
small angle
high HCT

9. CHARACTERISTICS OF A GOOD SMEAR
11.. TThhiicckk aatt oonnee eenndd,, tthhiinnnniinngg oouutt ttoo aa ssmmooootthh
rroouunnddeedd ffeeaatthheerr eeddggee..
22.. SShhoouulldd ooccccuuppyy 22//33 ooff tthhee ttoottaall sslliiddee aarreeaa..
33.. SShhoouulldd nnoott ttoouucchh aannyy eeddggee ooff tthhee sslliiddee..
44.. SShhoouulldd bbee mmaarrggiinn ffrreeee,, eexxcceepptt ffoorr ppooiinntt ooff
aapppplliiccaattiioonn..
Note: As soon as the drop of blood is placed on the glass slide, the smear
should be made without delay. Any delay results in anabnormal
distribution of the white blood cells, with many of the large white
cells accumulating at the thin edge of the smear.

10. COMMON CAUSES OF A POOR BLOOD
SMEAR
11.. DDrroopp ooff bblloooodd ttoooo llaarrggee oorr ttoooo ssmmaallll..
22.. SSpprreeaaddeerr sslliiddee ppuusshheedd aaccrroossss tthhee sslliiddee iinn aa jjeerrkkyy mmaannnneerr..
33.. FFaaiilluurree ttoo kkeeeepp tthhee eennttiirree eeddggee ooff tthhee sspprreeaaddeerr sslliiddee aaggaaiinnsstt
tthhee sslliiddee wwhhiillee mmaakkiinngg tthhee ssmmeeaarr..
44.. FFaaiilluurree ttoo kkeeeepp tthhee sspprreeaaddeerr sslliiddee aatt aa 3300°° aannggllee wwiitthh tthhee
sslliiddee..
55.. FFaaiilluurree ttoo ppuusshh tthhee sspprreeaaddeerr sslliiddee ccoommpplleetteellyy aaccrroossss tthhee sslliiddee..
66.. IIrrrreegguullaarr sspprreeaadd wwiitthh rriiddggeess aanndd lloonngg ttaaiill:: EEddggee ooff sspprreeaaddeerr
ddiirrttyy oorr cchhiippppeedd;; dduussttyy sslliiddee
77.. HHoolleess iinn ffiillmm:: SSlliiddee ccoonnttaammiinnaatteedd wwiitthh ffaatt oorr ggrreeaassee
88.. CCeelllluullaarr ddeeggeenneerraattiivvee cchhaannggeess:: ddeellaayy iinn ffiixxiinngg,, iinnaaddeeqquuaattee
ffiixxiinngg ttiimmee oorr mmeetthhaannooll ccoonnttaammiinnaatteedd wwiitthh wwaatteerr..

11. Examples of unacceptable smears
A: Blood film with jagged tail made from a spreader with achipped
.end
B: Film which is too thick
C: Film which is too long, too wide, uneven thickness and made on
.a greasy slide
.D: A well-made blood film

12. Examples of unacceptable smears

13. BIOLOGIC CAUSES OF A POOR SMEAR
11.. CCoolldd aagggglluuttiinniinn -- RRBBCCss wwiillll cclluummpp
ttooggeetthheerr.. WWaarrmm tthhee bblloooodd aatt 3377°° CC ffoorr 55
mmiinnuutteess,, aanndd tthheenn rreemmaakkee tthhee ssmmeeaarr..
22.. LLiippeemmiiaa -- hhoolleess wwiillll aappppeeaarr iinn tthhee ssmmeeaarr..
TThheerree iiss nnootthhiinngg yyoouu ccaann ddoo ttoo ccoorrrreecctt tthhiiss..
33.. RRoouulleeaauuxx -- RRBBCC’’ss wwiillll ffoorrmm iinnttoo ssttaacckkss
rreesseemmbblliinngg ccooiinnss.. TThheerree iiss nnootthhiinngg yyoouu
ccaann ddoo ttoo ccoorrrreecctt tthhiiss

14. Notes:
1. Although this is the easiest and most popular methods for
producing a blood smear, it does not produce a quality smear.
2. The WBCs are unevenly distributed and RBC distortion is seen at
the edges Smaller WBCs such as lymphocytes tend to reside in
the middle of the feathered edge.
3. 2. Large cells such as monocytes, immature cells and abnormal
cells can be found in the outer limits of this area.
4. 3. Spun smears produce the most uniform distribution of blood
cells.

15. SSLLIIDDEE FFIIXXAATTIIOONN &&
SSTTAAIINNIINNGG
LLEEIISSHHMMAANN''SS SSTTAAIINN

16. II- Fixing the films
 To preserve the morphology of the cells, films must be fixed as soon as
possible after they have dried.
 It is important to prevent contact with water before fixation is complete.
 Methyl alcohol (methanol) is the choice, although ethyl alcohol ("absolute
alcohol") can be used.
 Methylated spirit (95% ethanol) must not
be used as it contains water.
 To fix the films, place them in a covered staining jar or tray containing the
alcohol for 2-3 minutes. In humid climates it might be necessary to replace the
methanol 2-3 times per day; the old portions can be used for storing clean
slides.

17. III. Staining the film
Romanowsky staining:
Romanowsky stains are universally employed for staining blood
films and are generally very satisfactory.
 There are a number of different combinations of these dyes, which
vary, in their staining characteristics.
1. May-Grunwald-Giemsa is a good method for routine work.
2. Giemsa stain is thought to produce more delicate staining
characteristics.
Wright's stain is a simpler method.
4. Leishman's is also a simple method, which is especially suitable
when a stained blood film is required urgently or the routine stain
is not available (e.g. at night).
5. Field's stain is a rapid stain used primarily on thin films for malarial
parasites.

18. Principle
 The main components of a Romanowsky stain are:
… A cationic or basic dye (methylene blue or its oxidation
products such as azure B), which binds to anionic sites and
gives a blue-grey color to nucleic acids (DNA or RNA),
nucleoproteins, granules of basophils and weakly to granules
of neutrophils
… An anionic or acidic dye such as eosin Y or eosin B, which
binds to cationic sites on proteins and gives an orange-red
color to hemoglobin and eosinophil granules.
… pH value of phosphate buffer is very important.

20. Eosinophilic granules
Blue nucleus Basophilic granules

21. Staining procedure ( (Leishman’s stain
 Thin smear are air dried.
 Flood the smear with stain.
 Stain for 1-5 min. Experience will indicate the optimum time.
 Add an equal amount of buffer solution and mix the stain by
blowing an eddy in the fluid.
 Leave the mixture on the slide for 10-15 min.
 Wash off by running water directly to the centre of the slide
to prevent a residue of precipitated stain.
 Stand slide on end, and let dry in air.

22. SSttaaiinniinngg pprroocceedduurree
 TThhiinn ssmmeeaarr aarree aaiirr ddrriieedd..
 FFlloooodd tthhee ssmmeeaarr wwiitthh ssttaaiinn..
 SSttaaiinn ffoorr 11--55 mmiinn.. EExxppeerriieennccee wwiillll iinnddiiccaattee tthhee
ooppttiimmuumm ttiimmee..
 AAdddd aann eeqquuaall aammoouunntt ooff bbuuffffeerr ssoolluuttiioonn aanndd mmiixx
tthhee ssttaaiinn bbyy bblloowwiinngg aann eeddddyy iinn tthhee fflluuiidd..
 LLeeaavvee tthhee mmiixxttuurree oonn tthhee sslliiddee ffoorr 1100--1155 mmiinn..
 WWaasshh ooffff bbyy rruunnnniinngg wwaatteerr ddiirreeccttllyy ttoo tthhee cceennttrree ooff
tthhee sslliiddee ttoo pprreevveenntt aa rreessiidduuee ooff pprreecciippiittaatteedd ssttaaiinn..
 SSttaanndd sslliiddee oonn eenndd,, aanndd lleett ddrryy iinn aaiirr..

23. TTOOOO AACCIIDDIICC SSUUIITTAABBLLEE TTOOOO BBAASSIICC

24. CAUSES & CORRECTION
 TToooo AAcciidd SSttaaiinn::
11.. iinnssuuffffiicciieenntt ssttaaiinniinngg ttiimmee
22.. pprroolloonnggeedd bbuuffffeerriinngg oorr wwaasshhiinngg
33.. oolldd ssttaaiinn
 CCoorrrreeccttiioonn::
11)) lleennggtthheenn ssttaaiinniinngg ttiimmee
22)) cchheecckk ssttaaiinn aanndd bbuuffffeerr ppHH
33)) sshhoorrtteenn bbuuffffeerriinngg oorr wwaasshh ttiimmee

25.  TToooo AAllkkaalliinnee SSttaaiinn::
11.. tthhiicckk bblloooodd ssmmeeaarr
22.. pprroolloonnggeedd ssttaaiinniinngg
33.. iinnssuuffffiicciieenntt wwaasshhiinngg
44.. aallkkaalliinnee ppHH ooff ssttaaiinn ccoommppoonneennttss
 CCoorrrreeccttiioonn ::
11)) cchheecckk ppHH
22)) sshhoorrtteenn ssttaaiinn ttiimmee
33)) pprroolloonngg bbuuffffeerriinngg ttiimmee

26. PPEERRFFOORRMMIINNGG AA MMAANNUUAALL
DDIIFFFFEERREENNTTIIAALL AANNDD AASSSSEESSSSIINNGG RRBBCC
MMOORRPPHHOOLLOOGGYY

27. PPRRIINNCCIIPPLLEE
 WWhhiittee BBlloooodd CCeellllss..
11.. CChheecckk ffoorr eevveenn ddiissttrriibbuuttiioonn aanndd
eessttiimmaattee tthhee nnuummbbeerr pprreesseenntt ((aallssoo,,
llooookk ffoorr aannyy ggrroossss aabbnnoorrmmaalliittiieess
pprreesseenntt oonn tthhee ssmmeeaarr))..
22.. PPeerrffoorrmm tthhee ddiiffffeerreennttiiaall ccoouunntt..

28. PPRRIINNCCIIPPLLEE
 RReedd BBlloooodd CCeellllss,, EExxaammiinnee ffoorr::
11.. SSiizzee aanndd sshhaappee..
22.. RReellaattiivvee hheemmoogglloobbiinn ccoonntteenntt..
33.. PPoollyycchhrroommaattoopphhiilliiaa..
44.. IInncclluussiioonnss..
55.. RRoouulleeaauuxx ffoorrmmaattiioonn oorr aagggglluuttiinnaattiioonn
 PPllaatteelleettss..
11.. EEssttiimmaattee nnuummbbeerr pprreesseenntt..
22.. EExxaammiinnee ffoorr mmoorrpphhoollooggiicc aabbnnoorrmmaalliittiieess..

29. PROCEDURES
 Observations UUnnddeerr ××1100
11.. CChheecckk ttoo sseeee iiff tthheerree aarree ggoooodd ccoouunnttiinngg
aarreeaass aavvaaiillaabbllee ffrreeee ooff rraaggggeedd eeddggeess aanndd cceellll
cclluummppss..
22.. CChheecckk tthhee WWBBCC ddiissttrriibbuuttiioonn oovveerr tthhee
ssmmeeaarr..
33.. CChheecckk tthhaatt tthhee sslliiddee iiss pprrooppeerrllyy ssttaaiinneedd..
44.. CChheecckk ffoorr tthhee pprreesseennccee ooff llaarrggee ppllaatteelleettss,,
ppllaatteelleett cclluummppss,, aanndd ffiibbrriinn ssttrraannddss..

30. OOBBSSEERRVVAATTIIOONNSS UUNNDDEERR×× 4400XX :: WWBBCC
EESSTTIIMMAATTEESS
 UUssiinngg tthhee ×× 4400 hhiigghh ddrryy wwiitthh nnoo ooiill..
 CChhoooossee aa ppoorrttiioonn ooff tthhee ppeerriipphheerraall ssmmeeaarr wwhheerree
tthheerree iiss oonnllyy sslliigghhtt oovveerrllaappppiinngg ooff tthhee RRBBCCss..
 CCoouunntt 1100 ffiieellddss,, ttaakkee tthhee ttoottaall nnuummbbeerr ooff wwhhiittee
cceellllss aanndd ddiivviiddee bbyy 1100..
 TToo ddoo aa WWBBCC eessttiimmaattee bbyy ttaakkiinngg tthhee aavveerraaggee
nnuummbbeerr ooff wwhhiittee cceellllss aanndd mmuullttiippllyyiinngg bbyy 22000000..

31. U OBSERVATIONS UNNDDEERR ×× 110000:: PPLLAATTEELLEETT
EESSTTIIMMAATTEESS
11.. UUssee tthhee ooiill iimmmmeerrssiioonn lleennss eessttiimmaattee tthhee
nnuummbbeerr ooff ppllaatteelleettss ppeerr ffiieelldd..
22.. LLooookk aatt 55--66 ffiieellddss aanndd ttaakkee aann aavveerraaggee..
33.. MMuullttiippllyy tthhee aavveerraaggee bbyy 2200,,000000..
44.. NNoottee aannyy mmaaccrrooppllaatteelleettss..
 PPllaatteelleettss ppeerr ooiill iimmmmeerrssiioonn ffiieelldd ((OOIIFF))
11)) <88 ppllaatteelleettss//OOIIFF == ddeeccrreeaasseedd
22)) 88 ttoo 2200 ppllaatteelleettss//OOIIFF == aaddeeqquuaattee
33)) >>2200 ppllaatteelleettss//OOIIFF == iinnccrreeaasseedd

32. PLATELETS

33. OBSERVING AND RECORDING
NUCLEATED RED BLOOD CELLS (NRBCS)
 If 10 or more nucleated RRBBCC''ss ((NNRRBBCC)) aarree sseeeenn,,
ccoorrrreecctt tthhee
 WWhhiittee CCoouunntt uussiinngg tthhiiss ffoorrmmuullaa::
CCoorrrreecctteedd WWBBCC CCoouunntt ==
WWBBCC xx 110000//(( NNRRBBCC ++ 110000))
EExxaammppllee :: IIff WWBBCC == 55000000 aanndd 1100 NNRRBBCCss hhaavvee bbeeeenn
ccoouunntteedd
TThheenn 55,,000000×× 110000//111100 == 44554455..5500
TThhee ccoorrrreecctteedd wwhhiittee ccoouunntt iiss 44554455..5500..

34. MMAANNUUAALL DDIIFFFFEERREENNTTIIAALL CCOOUUNNTTSS
 TThheessee ccoouunnttss aarree ddoonnee iinn tthhee ssaammee aarreeaa aass
WWBBCC aanndd ppllaatteelleett eessttiimmaatteess wwiitthh tthhee rreedd cceellllss
bbaarreellyy ttoouucchhiinngg..
 TThhiiss ttaakkeess ppllaaccee uunnddeerr ×× 110000 ((ooiill)) uussiinngg tthhee
zziiggzzaagg mmeetthhoodd..
 CCoouunntt 110000 WWBBCCss iinncclluuddiinngg aallll cceellll lliinneess ffrroomm
iimmmmaattuurree ttoo mmaattuurree..
 RReeppoorrttiinngg rreessuullttss
AAbbssoolluuttee nnuummbbeerr ooff cceellllss//μμll == %% ooff cceellll ttyyppee iinn
ddiiffffeerreennttiiaall xx wwhhiittee cceellll ccoouunntt

35. OObbsseerrvviinngg ddiirreeccttiioonn::
OObbsseerrvvee oonnee ffiieelldd aanndd rreeccoorrdd tthhee nnuummbbeerr ooff WWBBCC
aaccccoorrddiinngg ttoo tthhee ddiiffffeerreenntt ttyyppee tthheenn ttuurrnn ttoo aannootthheerr ffiieelldd
iinn tthhee ssnnaakkee--lliikkeedd ddiirreeccttiioonn
**aavvooiidd rreeppeeaatt oorr mmiissss ssoommee cceellllss

36. NNOORRMMAALL PPEERRIIPPHHEERRAALL BBLLOOOODD SSMMEEAARR

37. LEUKOCYTOSIS
 Leukocytosis, a WBC above 10,000 is usually due to
an increase in one of the five types of white blood
cells and is given the name of the cell that shows the
primary increase.
1. Neutrophilic leukocytosis = neutrophilia
2. Lymphocytic leukocytosis = lymphocytosis
3. Eosinophilic leukocytosis = eosinophilia
4.Monocytic leukocytosis =monocytosis
5.Basophilic leukocytosis = basophilia

39. STAB NEUTROPHIL
 Diameter:12-16
 Cytoplasm : pink
 Granules: primary
secondary
 Nucleus: dark purple blue
 dense chromatin

40. BAND NEUTROPHIL

41. SEGMENTED NEUTROPHIL
 Diameter: 12-16
 Cytoplasm : pink
 Granules: primary
secondary
 Nucleus: dark purple blue
dense chromatin
2-5 lobes

42. SEGMENTED NEUTROPHIL

43. NEUTROPHILS.1
 Neutrophils are so named because they are not well
stained by either eosin, a red acidic stain, or by
methylene blue, a basic or alkaline stain.
 Neutrophils are also known as "segs", "PMNs" or
"polys" (polymorphonuclear).
 They are the body's primary defense against bacterial
infection.

44. Increased neutrophils count (neutrophilia)
1. Acute bacterial infection.
2. Granulocytic leukemia.
Decreased neutrophil count (neutropenia)
1. Typhoid fever
2. Brucellosis
3. Viral diseases, including hepatitis, influenza, rubella, and
mumps.

45. LEFT-SHIFT AND RIGHT-SHIFT OF NEUTROPHIL
 Normally, most of the neutrophils circulating in the
bloodstream are in a mature form, with the nucleus of the
cell being divided or segmented. Because of the segmented
appearance of the nucleus, neutrophils are sometimes
referred to as "segs.”
 The nucleus of less mature neutrophils is not segmented, but
has a band or rod-like shape. Less mature neutrophils -
those that have recently been released from the bone
marrow into the bloodstream - are known as "bands" or
"stabs".
 LLeefftt--sshhiifftt:: nnoonn--sseeggmmeenntteedd nneeuuttrroopphhiill >> 55%%
 RRiigghhtt--sshhiifftt:: hhyyppeerrsseeggmmeenntteedd nneeuuttrroopphhiill >>33%%

46. Segmented neutrophile Band neutrophil
Shift to left  Increased bands mean acute infection, usually bacterial.
Shift to right  Increased hypersegmented neutrophile.

47. EOSINOPHIL
 DDiiaammeetteerr:: 1144--1166
 CCyyttooppllaassmm :: ffuullll ooff ggrraannuulleess
 GGrraannuulleess:: llaarrggee rreeffrraaccttiillee,, oorraannggee--rreedd
 NNuucclleeuuss:: bblluuee
ddeennssee cchhrroommaattiinn
22 lloobbeess lliikkee aa ppaaiirr ooff ggllaassss

48. EOSINOPHIL

49.  The most common reasons for an increase in the
eosinophil count are
1. Allergic reactions such as hay fever, asthma, or drug
hypersensitivity.
2. Parasitic infection
3. Eosinophilic leukemia

50. BASOPHIL
 DDiiaammeetteerr:: 1144--1166
 CCyyttooppllaassmm :: ppiinnkk
 GGrraannuulleess:: ddaarrkk bblluuee ––bbllaacckk
oobbssccuurree nnuucclleeuuss
 NNuucclleeuuss:: bblluuee

51. BASOPHIL

52. Basophils
… The purpose of basophils is not completely understood.
… Basophile counts are used to analyze allergic reactions.
… An alteration in bone marrow function such as leukemia
may cause an increase in basophils.

53. LYMPHOCYTE
 DDiiaammeetteerr:: ssmmaallll 77--99
llaarrggee 1122--1166
 CCyyttooppllaassmm:: mmeeddiiuumm bblluuee
 GGrraannuulleess:: ssmmaallll
aaggrraannuullaarr
llaarrggee aa ffeeww
pprriimmaarryy ggrraannuulleess
 NNuucclleeuuss:: ddaarrkk bblluuee
rroouunndd
ddeennssee cchhrroommaattiinn

54. LYMPHOCYTE

55. LYMPHOCYTES.4
 Lymphocytes are the primary components of the
body's immune system. They are the source of
serum immunoglobulins and of cellular immune
response.
 Two types of lymphocytes:
1. B lymphocyte : Humoral immunity
2. T lymphocyte : Cellular immunity

56. Lymphocytes increase (lymphocytosis) in:
1.Many viral infections
2.Tuberculosis.
3.Typhoid fever
4.Lymphocytic leukemia.
A decreased lymphocyte (lymphopenia) count of less than 500
places a patient at very high risk of infection, particularly
viral infections.

57. MONOCYTE
 DDiiaammeetteerr:: 1144--2200
 CCyyttooppllaassmm :: ggrreeyy bblluuee
 GGrraannuulleess:: dduusstt--lliikkee
lliillaacc ccoolloorr ggrraannuulleess
 NNuucclleeuuss:: bblluuee
llaarrggee iirrrreegguullaarrllyy sshhaappeedd
aanndd ffoollddeedd

58. MONOCYTE

59. Diseases that cause a monocytosis
include:
•Tuberculosis
•Brucellosis
•Malaria
•Monocytic leukemia

60. NNOOTTEESS
11.. DDoo nnoott ccoouunntt cceellllss tthhaatt aarree ddiissiinntteeggrraattiinngg
• eeoossiinnoopphhiill wwiitthh nnoo ccyyttooppllaassmmiicc mmeemmbbrraannee aanndd wwiitthh
ssccaatttteerreedd ggrraannuulleess
• PPyykknnoottiicc cceellll ((nnuucclleeuuss eexxttrreemmeellyy ccoonnddeennsseedd aanndd
ddeeggeenneerraatteedd,, lloobbeess ccoonnddeennsseedd iinnttoo ssmmaallll,, rroouunndd cclluummppss wwiitthh
nnoo ffiillaammeennttss iinntteerrccoonnnneeccttiinngg))..
• ssmmuuddggee cceellllss
• BBaasskkeett cceellllss
•ssmmuuddggee cceellllss
•BBaasskkeett cceellllss

61. 2- Abnormal differentials
1. 200 Cell diff:
a. WBC > 15.0 (>20.0 for babies under 1 month and labor unit)
b. Three or more basophils seen.
2. If more than five immature WBC's are seen (or any blasts) let
someone else diff slide and average results.
3. Correct WBC for NRBC's if you seen ten or more NRBCs/100 WBC.
4. Always indicate number of cells counted on diff.
5. If any cell type is extremely elevated (such as bands, monos, or eos >
20) indicate that you are aware of the abnormality by circling or
checking on the card next to the results.

62. 3-Morphologic Changes Due To Area Of Smear
 Thin area- Spherocytes which are really
"spheroidocytes" or flattened red cells. True
spherocytes will be found in other (Good) areas of
smear.
 Thick area - Rouleaux, which is normal in such
areas. Confirm by examining thin areas. If true
rouleaux, two-three RBC's will stick together in a
"stack of coins" fashion..

63. tail body head

64. 4. A well-made and well-stained smear is essential to the accuracy of
the differential count. The knowledge and ability of the cell
morphologist is critical to high-quality results.
5. Before reporting significant abnormalities such as blasts, malaria or
other significant finding on a patient’s differential, ask a more
experienced tech to review the smear for confirmation. In clinical
settings where a pathologist or hematologist is present, the smear
is set aside for Pathologist Review.
6. Never hesitate to ask questions concerning morphology or the
identification of cells. The differential is one of the most difficult
laboratory tests to learn. In fact, learning about cells and their
morphology is a process that continues for as long as you perform
differentials.