Platelet Counts

Dr Kunal Sehgal, M.D.

Associate Consultant
Hematology Laboratory
Department of Lab Medicine
PD Hindua !ational Hospital and M"C
dr#unalsehgal$gmail.com
PD Hindua Hospital %ata memorial Hospital P&'M(", Chandigarh
Platelets ) Historical Perspecti*e

+,,-. Platelets recognised as distinct corpuscles ) 'talian pathologist

&iulio /i00o0ero

+123 . Manual Phase Contrast Microscopic Method using !eubauer

chamber . 'CSH . &old Standard +1,,.-44+

+152 .6-. Semi. Automated and 7ully Automated Counters

+1,+. Hydrodynamically focused 8hole blood aperture 'MP(DA!C(

counter

+1,2. 9P%'CAL Platelet Counts

(arly +114s. 7lo8 cytometric methods based on CD:+;5+

-44+. 7lo8 Cytometric "/C platelet "atio ) the ne8 'nternational reference
Method <'"M=
Manual Platelet Counts.
%he 9ld &old Standard

Laborious

%ime 'ntensi*e

Subecti*e

High 'nter. obser*er C>s of +4.-2 ?

Briggs et al. Continuing developments with the automated platelet count. Int. Jnl. Lab. Hem.2007,2,77!".
'nternational 7lo8 "eference
Method
%he !e8 &old Standard
"/C;Platelet "atio Method
Dual Platform Method
Absolute Platelet Count@
Platelet e*ents A "/C count
"/C e*ents <Automated Cell Analy0er=

ISLH Task Force, Am J Clin Pathol ++2, :54.:5:.<-44+=
C#
$"%&"
"/C
P
l
a
t
e
l
e
t
s
Peripheral /lood Smear
<Platelet count chec# only=

Platelets to be counted in a region 8here "/Cs and platelets

are 8ell dispersed.

Atleast +4 oil immersion fields to be counted <more in lo8er

counts=
A*erage no. of platelets in a field multiplied by +4444 is the
approBimate platelet count

Problems of Peripheral Smear
Platelet Chec#

Platelet Clumps

Platelet Satellitism on C/Cs

Poor Smearing

Highly subecti*e
Peripheral /lood Smear
<Platelet count chec# only=

(g D
a= +4 fields ) :2 platelets
A*g. plt per field is :.2
ApproBimate Platelet count@$.'("0000)$'000
b= -4 fields ) :4 platelets
A*g. plt per field is -
ApproBimate Plt count@2("0000)20000

A"%(7AC%S
Automated C/C Analysers

'mpedance principle

9ptical Principle
Counters count many more cells and hence
more reproducible results
'mpro*ed C.>. . typically less than 2?
'mpedance Principle

Coulter Principle or "esistance detection

method

Cells suspended in an elecrolyte solution

Change in electric impedance Eimpedance

signal

'mpedance signalE Directly proportional to

the *olume of the cell
C/C Histograms
!ormal Platelets histogram
&iant Platelets histogram
Problems 8ith 'mpedance
Counts
9ptical Principle
Two dimensional Light Scatter
%8o angles of laser ight scatter are measured
Light Scatter. -.3FC. *olume <plt si0e=
Light Scatter. 2.+2FC. refracti*e indeB <plt density=
"bc fragments ha*e a different "' as compared to
platelets and hence can be separated
Optical Fluorescence platelet counting
Si0e *s. 7luorescence plot <Polymethine Dye=
"/C fragments do not contain "!A 8hile giant
platelets and immature forms contain "!A and are
called reticulated platelets
%hese are easily separated from microcytic "/Cs
and fragments
Advantages of Optical Platelet
Counting
Microcytic RBC
Giant PLT
Optical Platelet Enumeration
CAS( S%GD'(S
Case Study +
Automated C/C .Platelet count ) +.42lacs
PS. many large platelet clumps
Chat do you doH
Peripheral Smear ) comment )
Platelets are seen in many clumps. Platelets are adeIuate
on smear <J+lac=. Kindly repeat C/C for accurate platelet
count if clinically indicated.
Case Study -
Automated C/C .Platelet count ) -.32lacs
PS. many platelet clumps
Chat do you doH
Peripheral Smear ) comment )
Platelets are adeIuate on smear. Platelets are also seen in
clumps.
Case Study 3 . *"%+,Blood #onor, ,ast Indian -rigin,
.ormal Hb and /BC, Impedance 0lt! "*$, 0latelet - 1"&2,
2orphologicall3! 2an3 4iant platelets
Case Study :. C/C Histogram
Case Study :. continuedK
Platelet Clumps in C/C &host Area
&host area in a case of platelet clumps
&host area in a normal C/C

6- year old male

Hemogram re*ealed thrombocytopenia

<2:,444;cmm=
Case Study 2
/ased on platelet histogram findings, a
peripheral smear eBamination 8as done

&iant platelets 8ere seen

Platelet clumps seen

%he sample contained adeIuate platelets,
ho8e*er 8e got spurious results on
automated analy0er
Peripheral smear sho8ing many
platelet clumps <+4B=.
(D%A induced Pseudothrombocytopenia
Citrated P/ Sample )Platelet count. -.32 lacs
Case Study 5
22;M A #no8 case of Acute Leu#emia
Hb !7.'g5 /BC! 2".' ("0* %ul 0latelet count! "6 ("0* %ul
/hat do 3ou do ne(t7
Peripheral smear chec#.

"ule out micro clots in sample

Loo# for fibrin strands and platelet clumps on slide

Do a peripheral smear estimation of platelet counts

/e a8are of the clinical decisions that depends on your result. i.e #no8 the
transfusion threshold le*els

Discuss case 8ith clinician if reIuired

Case study 6. Acceptable C.>.
Case Scenario +

7irst run, platelet count. -44444

Second run, platelet count ) +1-444

A difference of ,444. 's this AcceptableH Les. the
difference is only :?
Case Scenario -

7irst run platelet count. -:444

Second run platelet count ) +5444

A difference of ,444. 's this AcceptableH
.-! the di88erence is o8 **5 and will have a huge
clinical impact9
Any Muestions H