Beckman Coulter

Technologies
•Coulter Principle of counting and sizing
•Sweep Flow
•Pulse editing
•PLT curve fitting
•Coulter Histogram Differential
•Hgb Measurement
Prepared by
Muhammad A. Ghori
Current does not pass from
cells - they are insulator
Current does not pass from
cells - they are insulator
Electrolytes solutions allow
current to pass from them

Electrolyte
Like
ISOTON III
 Using this property of cells Wallace
Coulter invented the Coulter Counter in
late 50s
The Coulter Principle
RBC aperture
RBC aperture
The Coulter principle
A red cell
Red Blood Cell
passes through
RBC aperture

Sensing Zone

Oscilloscope
Oscilloscope
The Coulter principle
A red cell
Red Blood Cell
passes through
RBC aperture

Sensing Zone

Oscilloscope
Oscilloscope
The Coulter principle
A red cell
Red Blood Cell
passes through
RBC aperture

Sensing Zone

Oscilloscope
Oscilloscope
The Coulter principle
A White cell
Neutrophil
passes through
WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
The Coulter principle
A White cell
Neutrophil
passes through
WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
The Coulter principle
A White cell
Neutrophil
passes through
WBC aperture

Oscilloscope

Sensing Zone

Oscilloscope
 Passage of a RBC & Plt
through aperture

RBCs

+ -

PLT Cross section of Oscilloscope screen

RBC aperture
(60x50 micron)
 Passage of a RBC & Plt
through aperture

RBCs

+ -

PLT Cross section of Oscilloscope screen

RBC aperture
(60x50 micron)
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System without sweep flow

RBCs
recirculating

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
System with sweep flow

RBCs pushed
away by diluent

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
Diluent stream
System with sweep flow

RBCs pushed
away by diluent

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
Diluent stream
System with sweep flow

RBCs pushed
away by diluent

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
Diluent stream
System with sweep flow

RBCs pushed
away by diluent

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
Diluent stream
System with sweep flow

RBCs pushed
away by diluent

+ -
RBC

PLT

Oscilloscope screen

Sensing Zone
Diluent stream
Sweep Flow technology

RBCs swept away

from sensing
zone

Diluent stream
Pulse editing
Cell flows through centre of aperture

Good Pulse

Diluent stream
Pulse editing
Non central cell flow

Pulse to be
edited

Diluent stream
Coincidence Correction

Pulse to be
edited

Diluent stream
 Applying Thresholds
to separate Plt from RBC

20 fl

2 fl

Base line

Platelets
RBC of various
sizes
Coulter RBC histogram

Main RBC
population

RBC doublets

256 channel high resolution RBC histogram

display cell population data between 24 and 350 fl
 PLT histogram
and curve fitting
A fitted curve accurately reflecting
the collected data is displayed
between 2 and 35 fl

Raw data
Fitted Curve displayed up to 35fl

PLT histogram is based on 64 channels

 PLT curve fitting
The Platelet count is derived from the
number of cells under the fitted curve beyond
the 35fl display limit extending up to 70 fl

Raw data

Fitted Curve extend up to 70 fl to report very large PLT count

 PLT curve fitting
The curve fitting process allows more
accurate counts when platelets of
larger than 20 fl are present
 PLT curve fitting
The curve fitting process allows more
accurate counts when platelets of
larger than 20 fl are present

PLTs RBCs
Coulter WBC histogram

Lymphocyte Monocyte Granulocyte

256 channel high resolution WBC histogram

display cell population data between 35 and 450 fl
Coulter WBC histogram

Mono
Lymp

Neut

Baso Eos
 Hgb Measurement
In Beckman Coulter Instruments

WBC Bath Signal

ADC
Processor
HGB Lamp

Derivation
Absorbance= log 10 ( VR / Vs )

Where VR = Reference voltage

Vs = Sample voltage

HGB Sensor &

Sample PRE-AMP