Atlas Hematologi

8/11/2019 Atlas Hematologi

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1.Maturation of major leukocytes & others blood cells...........................3

2.Normal peripheral blood......................................................................5(1) rythrocytes and platelets (!oto1"2).....................................................................................5

(2) Neutrophil band #ranulocyte (!oto3"$).................................................................................5

(3) %olymorphonuclear neutrophil (!oto "1')............................................................................$

() osinophil (!oto 11&12)..........................................................................................................

(5) asophil (!oto 13"1$)............................................................................................................*($) +mall lymphocyte (!oto1"2')..............................................................................................,

() -ar#e lymphocyte (!oto 21"2)...........................................................................................1'

(*) Monocyte (!oto25"2*).........................................................................................................11

(,) %lasma cell (!oto 2,"32)......................................................................................................12

(1') typical lymphocyte (!oto 33"3*).....................................................................................13

3.rythoroblastic series of normal bone marro/...................................1(1)%roerythoroblasts (!oto3,"2)..............................................................................................1

(2)asophilic erythoroblast (!oto 3"$)..................................................................................15

(3)%olychromatic erythroblasts (!oto"5').............................................................................1$

()0rthochromatic erythroblasts (!oto51"5)...........................................................................1

.ranulocytic series in normal bone marro/.......................................1*(1)Myeloblast (!oto 55"$)........................................................................................................1*

(2)%romyelocyte (!oto$5"$*)....................................................................................................2'

(3)Myelocyte (!oto $,"2).........................................................................................................21

()Metamyelocyte (!oto3"$).................................................................................................22

5.0ther cells in normal bone marro/....................................................23(1)Mast cell (!oto&*)............................................................................................................23

(2)!at cell (!oto,&*')...............................................................................................................23

(3)Macropha#e (!oto*1"*3)......................................................................................................2

()0steoclasts (!oto*&*5)........................................................................................................25

(5)0steoblasts (!oto*$&*).......................................................................................................2$

($)eticulum cell(istiocyte) (!oto**&*,).................................................................................2$

()Me#akaryocyte(!oto,'&,1)...................................................................................................2

$.nomalies of erythrocytic series in peripheral blood.........................2*

(1)-eptocytes(!oto,2)................................................................................................................2*

1

4ontents


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(2)+pherocytes(!oto,3)..............................................................................................................2*

(3)lliptocytes(!oto ,)..............................................................................................................2*

()+tomatocytes(!oto ,5)...........................................................................................................2*

(5)+ickle cells(!oto,$)................................................................................................................2,

($)ar#et cells(!oto,)................................................................................................................2,

()nisocytosis(!oto,*)..............................................................................................................3'

(*)chinocytes(!oto,,)..............................................................................................................3'

(,)+chistocytes (6eratocytes) (!oto1'')....................................................................................3'

(1')canthocytes(!oto1'1)........................................................................................................3'

(11)ouleau7 formation (!oto 1'2)............................................................................................31

(12)ear drop cells (!oto 1'3)....................................................................................................31(13)asophilic stipplin# (!oto 1').............................................................................................32

(1)o/ell89olly bodies and %appenheimer bodies (!oto 1'5)...................................................32

.%reparation of smears and stainin# of peripheral blood.....................33(1)%reparation of +mear.............................................................................................................33

(2)+tandard +tains......................................................................................................................3

1):ri#ht +tain..........................................................................................................................3

2):ri#ht8iemsa +tain.............................................................................................................3

3)May8r;n/ald8iemsa +tain................................................................................................3

)iemsa +tain.........................................................................................................................3

2


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1.Maturation of major leukocytes & others blood cells

.Maturation of #ranulocytic series

1.Myeloblast 2"3.%romyelocyte (neutrophil) .Neutrophilic myelocyte

5.Neutrophil metamyelocyte $.Neutrophil band #ranulocyte

.Neutrophil se#mented #ranulocyte *",.%romyelocyte (osinophil)

1'.osinophilic myelocyte 11.osinophilic metamyelocyte

12.osinophilic band #ranulocyte 13.osinophilic se#mented #ranulocyte

1.%romyelocyte (asophil) 15.asophilic myelocyte 1$.asophilic metamyelocyte

1.asophilic band #ranulocyte 1*.asophilic se#mented #ranulocyte

.Maturation of Monocytic series

1. Monoblast 2.%romonocyte 3".Monocyte

4.Maturation of -ymphocytic series

1.-ymphoblast 2.%rolymphocyte 3.-ar#e lymphocyte .+mall lymphocyte

3


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$.Motosis) .%romyelocyte (!a##ot cell)

*.rape cell (includin# ussel bodies)

!

1.Neutrophil se#mented #ranulocyte sho/n basophilic cytoplasm

2.Neutrophil band #ranulocyte includin#


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2.Normal peripheral blood

(1) rythrocytes and platelets (!oto1"2)

!oto 1 !oto 2

rythrocytes are 8*m in diameter. =ie/ed throu#h the microscope> red cells are round.

+ince the actual shape> ho/eAer> is a biconcaAe disc> the central area is bri#hter. %latelets are 28

3(or 18m) in diameter. he cytoplasm /hich stains li#ht blue contains aBurophilic #ranules.

Nuclei are absent in both erythrocytes and platelets. he normal erythrocyteCplatelet ratio is

appro7imately 1582'D1> a helpful inde7 for reco#nition of thrombocytopenia and thrombocytosis.

(2) Neutrophil band #ranulocyte (!oto3"$)

!oto 3 !oto

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!oto 5 !oto $

(1)(3) and #ranulocytes (1)Myelocyte >(2)()+e#mented #ranulocytes

() +e#mented #ranulocyte>(5)Myelocyte (3)and #ranulocyte

Neutrophil band #ranulocyte haAe diameters about t/ice that of erythrocytes (about 15m)

he cytoplasm contains Ene #ranules stainin# li#htly oran#e8purple. he nuclear chromatin is

condensed> formin# coarse> dark purplish8red clumps. he nucleus is like a band of Aaryin#

/idths but the narro/ portions are not Eliform as in the mature neutrophil. @n normal blood>58

1'F of leukocytes are neutrophil band #ranulocytes.

(3) %olymorphonuclear neutrophil (!oto "1')

!oto !oto *

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!oto , !oto 1'

he siBe> cytoplasm and nuclear chromatin structure of polymorphonuclaer neutrophils are

+imilar to those of neutrophilic band #ranulocytes. o/eAer> the nucleus is se#mented into

three lobes by t/o Elaments. @n healthy adult> about 5'F of leukocytes are polymorphonuclear

neutrophils. Most polymorphonuclear neutrophils haAe t/o or three lobes.

() osinophil (!oto 11&12)

!oto 11 !oto 12

osinophils are sli#htly lar#er than neutrophils. he cytoplasm contains lar#e oran#e8red

#ranules.he nuclear chromatin structure is similar to that of neutrophils but the nucleus is

only se#mented into t/o s/ollen oAal lobes of eGual siBe by a Eliform stand. @n the normal

blood> about 3F of leukocytes are eosinophils. he leAel increases in cases of parasitic diseases>

such as ancylostomiasis> aller#ic reactions> such as bronchial asthma> and chronic #ranulocytic

leukemia.


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(5) asophil (!oto 13"1$)

!oto 13 !oto 1

!oto 15 !oto 1$

asophils are sli#htly smaller than neutrophils and characteriBed by the presence of lar#e>

darkblue stainin# #ranules in the cytoplasm and oAerlyin# the nucleus. +ince the #ranules sho/

metachromasia> the nameHbasophilH is really not correct. he nuclear chromatin structure and

shape of the nicleus often diIcult to obserAe clearly. @n normal blood> the percenta#e of

basophils is lo/> about '.5F of leukocytes. he #ranules contain histamine and heparin. he

#ranules may be diIcult toidentify in iemsa8stain smears> because they are too /ater8soluble

stain.

*


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($) +mall lymphocyte (!oto1"2')

!oto 1 !oto 1*

!oto 1, !oto 2'

+mall lymphocytes are sli#htly lar#er than erythrocytes(about 11Jm in diameter)and circular

in shape.he cytoplasm is scanty and clear blue> haAin# no #ranules. he nucleus stains deeply

due to condensationof chromatin. @n healthy adults> 3'85F of leukocytes are classed as

lymphocytes.

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() -ar#e lymphocyte (!oto 21"2)

!oto 21 !oto 22

!oto 23 !oto 2

-ar#e lymphocytes are round and t/ice as lar#e as the erythrocyte(about 15Jm in diameter).

he nucleus is lar#e and the cytoplasm plentiful. he nuclear chromatin structure and the

colorin# of thecytoplasm are similar to those of small lymphocytes> e7cept that a fe/

aBurophilic btanules are present in thecytoplasm.

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(,) %lasma cell (!oto 2,"32)

!oto 2, !oto 3'

!oto 31 !oto 32

%lasma cells are immuno#lobulin (@#> > < and ) producin# cells diKerentiated from 8

lymphocytes. hey haAe an aAera#e diameter of about 15Jm(1'2'Jm). he cytoplasm attains

deep blue> /hich indicates ih is ribosome rich and producin# immno#lobulin Ai#orously. +mall

Aacuoles are often ocserAed in the cytoplasm> indicatin# the presence of immno#lobulin. he

nucleus is eccentric. he lar#e ju7tanuclear li#ht area indicates /elldeAeloped ol#i apparatus.

he nucleus is round and relatiAely small. he nuclear chromatin is condensed and forms

clumps. No nucleolus is Aisible.

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(1') typical lymphocyte (!oto 33"3*)

!oto 33 !oto 3 !oto 35

!oto 3$ !oto 3 !oto 3*

!oto 33

typical lymphocytes are lar#e cells /ith diameters up to 2'Jm. he cytoplasm stains deep

blue. he nucleus is eccentric. here appear to be Aisible nucleoli. he shape of the cell is

similar to the plasma cell.

@f many atypical lymphocytes are found> the case can be dia#nosed as infectious

mononucleosis.

!oto 3*

/o cells /hich are similar to but sli#htly smaller than in the preAious foto are seen. hese are

not leukemia cells. +uch cells may appear not only in infectious mononucleosis> but also in Airal

hepatitis> rubella and hypersensitiAity to medicaments or serum.

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3.Erythoroblastic series of normal bone marrow

(1)%roerythoroblasts (!oto3,"2)

!oto 3, !oto '

!oto 1 !oto 2

%roerythroblasts are the lar#est cells of the erythroblast series. hey are around and 1

1,Jm in

diameter. he nucleus is lar#e and round. he nuclearCcytoplasmic (NC4) ratio is hi#h. he

cytoplasm is deeply basophilic (darker than that of myeloblasts) because of the abundance of

ribosomes and Ai#orous hemo#lobin synthesis. +eAeral bri#ht> less basophilic parts are ol#i

apparatus. he smaller ones in the bottom> /ith a honeycomb appearance> are mitochondria.

he nuclear chromatin structure is delicate and Enely #ranular. /o> three or more nucleoli are

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seen. 4haracteristically> they stain more deeply blue than the surroundin# re#ion (cf. in the

myeloblast> the nucleoli stain less blue than the surroundin# re#ion.)

(2)asophilic erythoroblast (!oto 3"$)

!oto 3 !oto

!oto 5 !oto $

asophilic erythroblasts are the second Aisible diKerentiation sta#e> follo/ih# proerythroblasts.

hey are sli#htly smaller (121Jm in diameter). s in proerythroblasts> the cytoplasm stains

dark blue. +ome bri#ht parts correspondin# to ol#i Bones and mitochondria are also seen

around the nucleus. he nucleus is lar#e. he nuclear chromatin structureis sli#htly coarser and

#ranular. No nucleous is Aisible. ransient forms bet/een proerythroblasts and basophilic

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erythroblasts may cause classiEcation diIculties. @f nucleoli are not eAident> ho/eAer> such

erythroblasts can be classiEed as basophilic.

(3)%olychromatic erythroblasts (!oto"5')

!oto !oto *

!oto , !oto 5'

%olychromatic erythroblasts are smaller bein# 1'15Jm in diameter. ecause of Ai#orous

hemo#lobin synthesis> the cytoplasm sho/s polychromatophilia> due to the blue stain of

basophilic ribosomes mi7in# /ith the oran#e8red of eosinophilic hemo#lobin. s the hemo#lobin

content increases> the color becomes more reddish. he nuclearCcytoplasmic ratio is smaller.

his means the siBe of the nucleus decreases more si#niEcantly than that of the cytoplasm. he

chromatin is more condensed> haAin# a so8called /heel8spoke appearance. %olychromatic

erythroblasts are normally the most numerous representatiAes of the erythroblast series. he

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ratio of proerythroblasts to basophilic erythroblasts to polychromatic erythroblasts to

orthochromatic erythroblasts is about 1D,D*'D1' in normal bone marro/.

()0rthochromatic erythroblasts (!oto51"5)

!oto 51 !oto 52

!oto 53 !oto 5

0rthochromatic erythroblasts are much smaller (*12Jm)> but sli#htly lar#er than mature

erythrocytes. he nuclear chromatin is markedly condensed. @t often attains deeply /ithout

e7hibitin# any structure> althou#h some structure may be obserAed on occasion. he

nuclearCcytoplasmic ratio decreases further. .s seen here> the cytoplasm possesses the same

color as mature erythrocytes. +trictly speakin#> orthochromatic erythroblasts under#o e7trusion

of the nuclei to become reticulocytes that are polychromatic because of the small amount of

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ribosomes persistin#. he ability to diAide is lost at the orthochromatic erythroblast sta#e.

4.ranulocytic series in normal bone marrow

(1)Myeloblast (!oto 55"$)

!oto 55 !oto 5$

!oto 5 !oto 5*

!oto 5, !oto $'

1*


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!oto $1 !oto $2

!oto $3 !oto $

Myeloblasts are the most immature cells of the #ranulocytic series that can be identiEed

microscopically. hey are 122'Jm in diameter and round. he cytoplasm is relatiAely scanty

and stains blue (li#hter than that of proerythroblasts) but more intensely at the mar#in of the

cell. lthou#h there are not many #ranules> a fe/ Ene aBurophilic #ranules may be obserAed in

some cases. he ju7tanuclear li#ht area is in #eneral not so distinct as in proerythroblasts. he

nucleus is #enerally round> haAin# a delicate> lace8like chromatin structure. here are 2 5

nucleoli that are bri#hter than the surroundin#s> in contrast to those in proerythroblasts. +ince

the ori#in of monocytes and bone marro/ #ranulocytes is the same> it may be impossible to

distin#uish monoblasts from myeloblasts. +ince only

a fe/ promonocytes are present in normal marro/> it is diIcult to identify them. Monocytic

leukemia monoblasts and promonocytes can be discriminated by usin# esterase stain.

1,


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(2)%romyelocyte (!oto$5"$*)

!oto $5 !oto $$

(1)%romyelocyte> (2)Metamyelocyte>

(3)"()+e#

(5)($)Myelocyte

!oto $ !oto $*

(1)%romyelocyte> (2)Myelocyte> (3)and form

%romyelocytes are lar#er than myeloblasts (1223Jm in diameter). he nuclearCcytoplasmic

ratio decreases. his means that the cytoplasm is more plentiful> the stainin# Aaryin# from that

of the myeloblast to the li#ht blue of the myelocyte. 4oarse aBurophilic #ranules (%rimary

#ranules) are conspicuous. hey also oAerlie the nucleus in some cases. he nucleus is round or

oAal and tends to lie eccentrically in the cytoplasm. he nuclear chromatin structure is still

delicate and nucleoli are Aisible. @n some cases> ho/eAer> the chromatin is as condensed as in

myelocytes. @t is said that the appearance of speciEc #ranules (secondary #ranules) can be

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obserAed at this sta#e by electron microscopy and they can be classiEed as neutrophi>

eosinophil> or basophil. +uch discrimination is impossible by li#ht microscopy.

(3)Myelocyte (!oto $,"2)

!oto $, !oto '

!oto 1 !oto 2

(1)&(2)Myelocyte> (3)&()Metamyelocyte (1)&(2)Myelocytes

Neutrophil myelocytes are smaller than promyelocytes (122'Jm). he cytoplasm is still

bluish but turns pink as the cell matures. No coarse aBurophilic #ranules are Aisible (althou#h

electron microscopy reAeals that primary #ranules remain> they no lon#er stain). +cattered Ene

neutrophil (secondary) #ranules are found. he nucleus is smaller but still round. he chromatin

is obAiously condensed and stains deeply. he mar#ins of the chromatin clump are distinct. No

nucleolus is Aisible. 4ells after the myelocyte sta#e haAe no diAision potential.

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()Metamyelocyte (!oto3"$)

!oto 3 !oto

(1)Metamyelocyte> (2)and form (1)&(2)Metamyelocytes> (3)and form

!oto 5 !oto $

(1)&(3)Metamyelocyte> (2)and form

he cytoplasm of neutrophil metemyelocytes is similar to or sli#htly less than that of

myelocytes (12 1*Jm). he color is pink and close to that in band forms and

polymorphonuclear neutrophils. feature useful for discriminatin# neutrophil metamyelocytes

from myelocytes is that the nuclei are concaAe and kidney8shaped. he nuclear chromatin is

much more condensed.

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!."ther cells in normal bone marrow

(1)Mast cell (!oto&*)

!oto !oto *

Numerous relatiAely coarse #ranules stainin# deep purple because of metachromasia occupy

the cytoplasm. +uch #ranules> often resultin# in unclear contours oAerlie the round nucleus.

Mast cells can be identiEed easily eAen under lo/ ma#niEcation> because the /hole cell is seen

as a black8purple clump. Mast cells are round or spindle8shaped. +ome cells haAe a lon# major

a7is. ranules contained in mast cells are similar to those in basophils at a #lance but they

diKer from each other in that the former are insoluble in methanol and thus are clearly obserAed

by iemsa stain. he ori#in of mast cells is unkno/n. lthou#h mast cells are distributed not

only in the bone marro/ but also throu#hout the body> none are obserAed in the peripheral

blood. he #ranules contain heparin and histamine.

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(2)!at cell (!oto,&*')

!oto , !oto *'

!at cells are Aery lar#e (about 1''Jm in diameter). hey haAe a round or oAal nucleus that may

be located centrally or peripherally. he nucleus has a sparse chromatin structure. No nucleolus

is Aisible.

+ince fat in the cytoplasm escapes durin# smear preparation> the cytoplasmic area may appear

empty> only a folded membrane bein# obserAed. @n bone marro/ hyperplasia fat cells are said

to contain less fat.

hey are star8shaped by becomin# trapped by the proliferatin# haemopoietic cells. s a result>

they can be diIcult to identify.

(3)Macropha#e (!oto*1"*3)

!oto *1 !oto *2

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!oto *3

Macropha#es Aary in siBeL some are as lar#e as monocytes> others are like me#akaryocytes.he nucleus is round or oAal> and appears bri#ht due to the sparse> spon#e8like chromatin

structure. 0ne or t/o blue8stainin# nucleoli may be obserAed. he cytoplasm is #enerous>

stainin# li#ht blue. he cell is irre#ularly shaped /ith lon# protrusions and an often indistinct

border. +ome macropha#e may contain pha#ocytosed cells or cell debris (a pha#ocytosed

erythroblast is indicated by the arro/).

@t is considered that the ori#in of monocytes and macropha#es is the same and that monocytes

become macropha#es /hen they enter the tissues from the peripheral blood. herefore> cells

considered as intermediate forms may be obserAed in some cases. @t is no/ common to use the

termmacropha#eas a synonym for histiocyte. @n other /ords> macropha#es belon# to

the monocyteChistiocyte (monocyteCmacropha#e) series. s a result> it has become necessary to

reclassify and considerably modify deEnitions of cells in the traditional reticuloendothelial

system.

()0steoclasts (!oto*&*5)

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!oto * !oto *5

0steoclasts are Aery lar#e cells of $'81''Jm or mare diameter. he cytoplasm stains a li#ht

blue and numerous Ene purple8red aBurophilic #ranules are Aisible. hey are distin#uished from

ma#akaryocytes by their round or ellipsoid nuclei. he nucleus is about 15Jm in diameter> /ith

one> or in some cases> t/o or three deeply stainin# nucleoli. he nuclear chromatin structure is

relatiAely delicate and #ranular.

0steoclasts are not uncommon in the marro/ of infants because oh the Ai#orous bone

remodelin# that is occurrin#. hey are less common in adults bein# seen only in patients /ith

myeloma and metastatic cancer.

(5)0steoblasts (!oto*$&*)

!oto *$ !oto *

0steoblasts look like plasma cells at a #lance> but are much lar#er than the latter (about 2'8

'Jm).

hey are round or spindle8shaped /ith irre#ularly shaped contours. lthou#h the cytoplasm

stains blue in #eneral> li#hter parts (chromophobic areas) may be obserAed in some cases. +uch

parts are not in the ju7tanuclear li#ht area but a/ay from the nucleus. small number of

aBurophilic #ranules are Aisible. he periphery of the cell has sli#htly irre#ular contours.

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4haracteristically> the nucleus is round or oAal> and located eccentrically. he nuclear chromatin

structure is delicate and lace8like. ypically> one> or> in some cases> t/o or three blue stainin#

nucleoli are obserAed. 0steoblasts can be obserAed in persons /ith actiAe ne/ bone marro/ but

rarely in adults. o/eAer> they can be obserAed alon# /ith osteoclasts in disease associated

/ith ne/ bone formation.

($)eticulum cell(istiocyte) (!oto**&*,)

!oto ** !oto*,

he cell sho/n here is lymphoid histiocyte /ith an indistinct periphery. %ha#ocytosis is not

obserAed. his cell used to be called the lymphoid reticulum cell.

()Me#akaryocyte(!oto,'&,1)

!oto ,' !oto ,1

!oto ,'

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Me#akaryocytes are considerably lar#er than prome#akaryocytes. his foto sho/s that the

cytoplasm contains a lar#er number of purplish8red #ranules. +ome sli#ht platelet producin#

potential is obserAed in the cytoplasm belo/ the nucleus.

!oto ,1

his me#akaryocyte is much lar#er (5'8$'Jm in diameter). he nucleus is irre#ularly shaped.

he nuclear chromatin is condensed. he cytoplasm is plentiful> /ith platelets bein# produced

Ai#orously. platelet demarcation system is formed in the cytoplasm of me#akaryocytes> part of

/hich is

#.$nomalies of erythrocytic series in peripheral blood

(1)-eptocytes(!oto,2)

he bri#ht central area is lar#er> /hile the cell is sli#htly smaller than normal.

nisocytosis is more conspicuous. +uspect iron deEciency anemia Erst.

(2)+pherocytes(!oto,3)

hese are small> circular> deeply stained (thick) erythrocytes. :hen present> suspect

hereditary spherocytosis or autoimmune hemolytic anemia

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!oto ,2 !oto ,3

(3)lliptocytes(!oto ,)

llipsoid or oAal erythrocytes are numerous. :hen present in such numbers there must be

a hi#h suspicion of hereditary elliptocytosis.

()+tomatocytes(!oto ,5)

hese cells are characteriBed by their narro/ slitlike li#hter8stained central areas.

he dia#nosis is stomatocytosis. +tomatocytes look like this as a result of smearin#. :henobserAed by scannin# electron8microscopy> stomatocytes are shaped like a bo/l.

!oto , !oto ,5

(5)+ickle cells(!oto,$) lon#ated erythrocytes lookin# like a sickle /ith sharp8pointed ed#es are conspicuous. he

dia#nosis is sickle cell anemia (b+ disease).

($)ar#et cells(!oto,)

ar#et cells are hypochromic erythrocytes lookin# like a tar#et because they stain

they are like a thin cup or salad bo/l. :hen present> thalassemia major or

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obstructiAe jaundice should be suspected. ar#et cells are also found in b +4

such an

appearance su##ests hereditary (con#enital) nonspherocytic hemolytic anemia. ed cell

enByme deEciencies> such as pyruAate kinase deEciency> or unstable hemo#lobin disease

should be considered.

(*)chinocytes(!oto,,)

he arro/s in the !oto indicate t/o small (probably dehydrated) contracted erythrocytes.

chinocytes are characteristically found in postspenectomy patients /ith enByme

anomalies of the erythrocyte #lycolysis system. o/eAer> note that a fe/ echinocytes may

be obserAed eAen before splenectomy. chinocytes are also called crenated cellsand

may be artifacts> for e7ample> /hen dryin# a smear is delayed before stainin#.

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!oto ,* !oto ,,

(,)+chistocytes (6eratocytes) (!oto1'')

+chistocytes are also called red cell fra#mentsorburr cells. %oikilocytosis due to

mechanical red cell fra#mentation is conspicuous. he shape Aaries from a trian#le or

helmet to a small sphere. his smear is from a patient /ith hemolytic uremic syndrome.

(1')canthocytes(!oto1'1)

canthocytes (or spur cells) /hich are irre#ularly shaped poikilocytes /ith Aariably lon#

protrusions> are conspicuous. hey are found in patients /ith cirrhosis accompanied by

plasma lipid alteration. +uch erythrocytes may be found in some patients /ith chorea8like

hereditary neuropathy. canthocytes can be clearly distin#uished from echinocytes.

!oto 1'' !oto 1'1

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(11)ouleau7 formation (!oto 1'2)

his looks like a pile of coins. ouleau7 formation is characteristic of hyper#lobulinemia.

+uspect myeloma or macro#lubulinemia> /hen present.

(12)ear drop cells (!oto 1'3)

he arro/s in the !oto 1'3 indicate tear drop cells. +uspect myeloEbrosis.

!oto 1'2 !oto 1'3

(13)asophilic stipplin# (!oto 1')

he erythrocytes indicated by the arro/s haAe prominent basophilic stipplin#. +uspect

acute lead poisonin# or pyrimidine858nucleotidase deEciency of erythrocytes. asophilic

stipplin# may be also obserAed in other disease> such as mali#nant anemia> unstable

hemo#lobin disease> thalassemia and myelodysplastic syndrome (M no such Aacuoles are usually obserAed. o/ell89olly bodies are nuclear residues

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that are round /ith 1 Jm diameter and stain deep purple. %appenheimer bodies contain

iron and therefore appear blue8#reen> /hen stained by iron stain. his means that

erythrocytes /ith %appenheimer bodies are siderocytes. oth types of inclusions tend to

appear after splenectomy. o/ell89olly bodies are often seen in dyserythropoiesis> for

e7ample> pernicious anemia> osteomyelodysplasis syndrome and thalassemia.

!oto 1' !oto 1'5

%.reparation of smears and stainin' of peripheral blood

(1)%reparation of +mear

o prepare smears> #lass slides are cleansed by soakin# in mild deter#ent> /ashin#>

immersin# in anhydrous alcohol for a half day or more> and then /iped thorou#hly

(alternatiAely clean #lass slides can be obtained commercially) L a coAer slip

(1*mm2mm> '.mm thickness) or #lass spreader is used to prepare the smear L drop $8

Jl of the blood containin# an usin# the tip of a #lass rod> capillary or needle (for blood collected

from a En#ertip or earlobe> place one drop of the blood directly on to the #lass spreader) L

place the #lass slide on a Elm Oat surface> and spread the blood so that it is in contact /ith

the entire ed#e of the spreader L immediately> position the spreader at an an#le of 3'

de#rees a#ainst the slide #lass and slide it in the direction the arro/ sho/n in !i#ure 1 /hile

33


34/35

pushin# it li#htly to smear the blood. @mmediately after smearin#> dry the blood /ith a fan

or hair dryer (at room temperature).

!or better Guality of erythrocyte morpholo#y> allo/ the smear to stand for 3' minutes to 1

hour before E7in# and stainin#.

(%0@N+)

@f the smear is too short> the correct re#ion for e7amination is limited. lood should be

smeared in such a /ay that the tail is about 2cm from the end of the slide. he smaller the

an#le bet/een the slide and the spreader and slo/er the speed of spreadin#> the thinner

the blood smear. @t is recommended that blood from a patient /ith anemia be smeared at a

lar#er an#le (358'rather than 3') and at a hi#her speed and that blood from a patient

/ith polycythemia be smeared at a smaller an#le and at a slo/er speed. automated deAices for smearin# and stainin# blood are aAailable. one marro/

smears should> ho/eAer> continue to be prepared by hand.

ently dra# the #lass to/ard you in the direction of the arro/ (1) spreadin# the blood oAer

an area eGual to the #lass spreader> then /ith a uniform speed slide the #lass in the

direction of the arro/(2)

(2)+tandard +tains

1()ri'ht *tain %lace ten drops of :ri#ht stain (a mi7ture of methylene aBure and eosin in methanol for

stainin# and E7ation) on to a horiBontally placed smear. llo/ the smear to stand for 1 to 3

minutes. :hen the room temperature is hi#h and humidity lo/> the preparation should be

coAered to preAent eAaporation of the stain. %lace ten drops of 1C15 molC- phosphate buKer

solution (%h $.). Psin# a #lass rod held horiBontally in the hand #ently stir the buKer solution

/ith the stain takin# care not to dama#e the smear. lternatiAely> blo/ #ently to mi7 the buKer

and stain solutions. llo/ the smear to stain for about 5 minutes (or a sli#htly lon#er time> /hen

the smear includes a lar#er number of leukocytes or is bone marro/). :ash the smear in a /eak

3

3'

lass spreader

+lide #lass

+uitable Aie/in#

+i'ure 1

lood


35/35

stream of /ater in such a /ay that stain residuals are remoAed.

2()ri'ht,iemsa *tain he iemsa stain is a mi7ture of aBure (an eGuimolar mi7ture of methylene blue and

methylene aBure) and an eGuimolar mi7ture of aBure and methylene blue eosinoate. he

stainin# procedure is the same as for the :ri#ht stainin# method> e7cept that the stainin# time

is shorter> about 3 minutes. :ash the smear /ith /ater and drain oK the /ater. compared

/ith that stained by :ri#ht8iemsa stain.

4(iemsa *tain !i7 the smear for 182 minutes in methanol and stain /ith the diluted iemsa solution for 2'83'

minutes. :ash the smear /ith /ater and dry. +mears stained only by the iemsa method>

produce poor stainin# of #ranules /hereas those stained by only the :ri#ht stain haAe poor

nuclear deEnition. s a result either May8r;n/ald8iemsa or :ri#ht8iemsa stains are

recommended. he @nternational 4ouncil for +tandardiBation in aematolo#y (@4+)

recommends a stain techniGue usin# a mi7ture of puriEed aBure and eosin

Atlas Hematologi

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