Hematology Analyzer Engineer

Training

Manufacture standard, manufacture reliance

 Training schedule

1、Use of Hematology Analyzer

2、Basic knowledge of Blood
2、Hematology Analyzer’s principle and
development.
3、Clinic operation, parameter and test performance.
4、Parameter introduction and instrument
installation.
 What is Hematology Analyzer
Hematology analyzer : It use for blood clinic
diagnose.

Hematology analyzer function.

Test item：HGB(Hemoglobin content)、RBC( red
blood cell count), WBC (white blood cell count)
and WBC label.
Basis blood knowledge
The compose of whole blood
 Blood corpuscle sort and modality

WBC RBC PLT

life-span：7days 120days 8days
diameter：20um 6.7~7.7um 2~30fl
Function :Anti-infection carry oxygen
Immunization metaboly coagulation
WBC
Granulocyte（75%） monocyte （5%） lymphocyte（25%）
Neutrophile (65~70%)
Eosinophile(0.5%~1.5%)
Basophile (0~1%)
Blood corpuscle compose
 Three group definition
Three group is that separate the cell of WBC：
lymphocyte , middle cell( Basophil、Eosinophil、monocyte）、
granulocyte。

WBC histogram ：

From left to right, lymphocyte, single karyon cell and grain cell, the
relative volume range are 35-90fl、90-160fl、160-450fl。
 RBC

• The HGB is included in the RBC, the HGB make the blood
red. HGB combine with the oxygen and release the carbon
dioxide, which make the body breathe in the fresh oxygen
and breathe out the carbon dioxide. In human body, the
quantity of RBC is more than other blood cell,
Adult man 4.29-5.23X
Adult woman 3.83-4.83X

The diameter of RBC is between 6 nanometer and 9

nanometer, the average is 7.2 nanometer.
 WBC

• WBC has karyon，lick up pathogen

invading body, which can protect the human
body prevent the invasion of microbe and
other infection, so that, the RBC goes by the
name of human body kavass.
• Healthy people RBC quantity is 4-10X
• The diameter is between 7 nanometer and
25 nanometer.
 PLT

PLT has not karyon with small and irregulary

body, it help the blood coagulate and gore
shrink.
Healthy people PLT quantity is100-300X
The diameter is between 2 and 3 nanometer
 HGB

HGB is one kind of ferriferous composite albumen,

HGB is basis of RBC. It displays cardinal red when it
combine with oxygen flowing through the lung.
The cardinal red is feature of artery blood, it displays
dark amethyst which is the feature of vein blood.

Adult man HGB concentration in 100 milliliter: 12-16g，

Adult woman HGB concentration in 100 milliliter: 11-15g。
RT-7600
 Main feature

1、RBC trisection hive off, 20 item test parameter and 3

histogram.
2、10.4 inch clear color big screen, touch screen , mouse
and keyboard operation enabled.
3、9.8ul Aspiration sample volume ，Whole blood, Pre-
dilute Peripheral blood
4、50000 test results, USB port for data transfer.
5、Test rate: 60test/hr
6、RBC/PLT/WBC histogram can use the drift terminus
for accuracy result.
Hematology Analyzer’s development
and principle
 Early blood corpuscle counting
analyzing method

Manual counting:
Classical microscope counting
Use the microscope counting cell and
analyzing
Procedure of counting by manual

1、Blood corpuscle counting（RBC、WBC、PLT

etc counting under the microscope）
2、Quantificational HGB
3、Sorting of WBC
4、Other: blood corpuscle volume rate, RBC
modality, counting etc. Eosinophil counting
 Current blood corpuscle counting

• 1950~80 year (semi-auto): need to dilute the

sample by manual then test by instrument,
high carry over, high random error.

• After 1980 year（auto）：the blood is

aspirated into instrument , auto-dilute , auto
hemolytic agent adding, auto-counting. The
high precision and accuracy.
By manual instrument counting
Manual Instrument
High random error, counting The test speed is increasing
equipment system error and with the technology improve.
method error. Counting need The test item increase, high
more time and effort ,results speed and high accuracy is
with low precision and the feature of the instrument.
accuracy. Supply more experiment data
with clinic.
Fundamental of the hematology analyzer
 Electronic impedance variation principle
Blood corpuscle is one kind of nonconductor, put some corpuscle in
liquid, which has no effect on the resistant of the whole liquid
because the small size of the cell.
However, place some corpuscle in certain small size liquid circuit,
which will generate variation on the resistance.
Often, we let the blood corpuscle pass though the micro-aperture ,then
test the resistance variant of the relative electrical field.
The figure showing:
 Different pulse

If the conduct liquid has been insert the constant electric current (current remain
unchanged), the voltage is increase between the two electrode, generate the electric
pulse is proportion to the resistance (impedance)

When specific quantity volume liquid with blood corpuscle pass through the micro-aperture,
one serial pulse generate, counting the pulse can get the numerical value of the blood
corpuscle.
Since the difference diameter of blood corpuscle, the impedance is different, then the pulse
is different, which can grouped the blood corpuscle.
 Counting principle 1

An electronic micro-field is generate around the micro-

aperture in which the blood cells pass through . The cells
create a resistance in the electronic field as they pass the
calibrated micro-aperture. This in-turn cause an electronic
pulse to be generate, which is amplified, measured, and
then mathematically calculated to create a numerical value.
First, the volume dilute blood sample is diluted in an
electronic diluent (electronic current conducting filed)
mixed, then pulled through a calibrate micro-aperture.
There are two electrodes that are placed on each side of the
aperture and a constant electronic current pass between
them.
 Counting principle 2

As the blood cell pass through the aperture, they create

resistance (impedance) in the electronic field between the
two electrodes ,since the current is constant and remain
unchanged, the large the cell is, the “more” resistance it
has. The small the cell is, the “less” resistance it has. The
voltage which measure the cell is proportional to the
voltage will be. The small the cell, the lower voltage will
be.
These electronic voltage vary in pulse as cells pass through
the aperture . The pulse are then changed according to
pulse size. The pulse size are then threshold grouped, then
mathematically calculate to create a Numerical value for
the determination of RBC’s and PLT’s
 Counting principle 3

• A certain amount of cells will pass through

the calibrated micro-aperture within a
specific time frame. They are then measure
by pulse height, threshold; grouped by size,
and mathematically calculated along with
the calibration coefficient to give the final
numerical value for both RBC’s and PLT’s
 Counting principle 4
As we all know ，the RBC number in human body is unit，WBC is
unit, that is to say the RBC is 1000 times to the WBC. When counting RBC,
the WBC can be included, since error is 1/1000，which has no effect on the
clinic diagnose.。
The diameters of the RBC, WBC and PLT are different:
RBC: 6-9 um WBC: 7-25 um PLT: 2-3 um
They generate different pulse when pass through the micro-aperture, the pulse of
WBC is more, then the RBC’s and PLT’s, the figure showing
 WBC counting

Before counting, the lysing reagent is added

to dilute blood in WBC chamber, the lyse
reagent contains potassium ferricyanide and
potassium cyanide, (most of reagent without
cyanide), the lysing reagent break down the
RBC cell membrane and release the
hemoglobin within the RBC, then count the
WBC.
 HGB testing

When the RBC cell membrane is break down, the

HGB inside will be released, it diffused in the
liquid.
The Hemoglobin combines with potassium cyanide
to form a chromogenous cyanmethmoglobin
compound, this chemical compound is measured
by spectrophotometry through the optical pathway
in the WBC chamber, the light wavelength of
measurement is at 550 nm.
HGB= Log (blank value/sample value)x the
calibration coefficient
 Superposition counting loss

In the perfect condition, the cell pull through

the micro-aperture one by one.
In the practical operation, there will be two or
three cell pull through the aperture at the
same time, though they can generate one
high pulse, only one cell is counted, which
lead to the loss in counting cell.
It is defined that superposition counting loss.
 Superposition loss remedy

In order to remedy the loss，some instrument have circuit for

revising or software revising.
The revising according to the posson law:
when counting number lower than 8000 unit, no revising.
When counting number is between 8000 and 38000，100
unit are add to per 900 unit.
When counting number is greater than 38000 ,200 unit are
add to 800 unit.
The revised counting number approach to the clinic real result.
 Micro-aperture principle 1
Micro-aperture is the key part of the signal transfer, which is the aperture on slice
of the ruby or sapphire，the slice is sticked to the glass or steel .

The diameter is about 75um or 100um. The aperture is near to the tube side of
bottom, some locate on the bottom.
100um aperture can pass and test the 2-40um particulate.

The diameter has an effect on the impedance value of the electrolyte between the
electrode A and B. Commonly, the 100um aperture in clinic brine，which is
about 15kΩ resistance.

When the aperture is blocked, please do not use the tinsel to get through,
otherwise, it will make the aperture coarse or destroyed the aperture.

The micro-aperture is precious part, please keep it well.

Micro-aperture principle 2
 Blood corpuscle histogram
The hematology analyzer also supply the volume
distribution histogram of RBC,WBC and PLT. The
histogram x-axis represent the size of the cell (pulse
height), Y-axis represent the number of the cell (pulse
frequency).
The histogram base on the pulse signal size and
frequency, plotted by the computer system.
The histogram supply not only the intuitionistic
test result, but instrument working condition.
Blood corpuscle pulse and
histogram
WBC Histogram
 Description of WBC histogram

The hematology analyzer group the cell from

35 ～ 450fl according to the size of the cell.
First group: 35 ～ 90fl small cell section，mainly
the lymphocyte
Second group: 90 ～ 160fl middle cell section,
mainly the monocyte, eosinophile, basophile and
neutrophile .
Third group:160fl-450fl big cell section, mainly
the granulocyte.
 Abnormal histogram of WBC

The histogram belong to jaundice patient, the lipid within

RBC increase, the protein within RBC decrease, which
contradict the lyse reagent, the lyse reagent has difficult in
breaking down RBC.
Histogram of RBC
 Description of RBC histogram
Hematology analyzer analyze RBC on 36 ～ 360fl, X-
axis represent the size (pulse height) of the cell, Y-axis
represent the number (pulse frequency) of the cell.
Normal RBC distribute in the range of 50 ～ 200fl，
there are two cell group on the histogram, one is from
50fl to 125fl, which is the symmetry distribution curve.
On the right side is the cell in range of 125 ～
200fl,which is the big RBC and netting shape RBC.
when the size of RBC change, the distribution will move
left or right, sometime the double apices, or the fundus
of curve widen.
 Abnormal histogram of RBC

Double apices on the curve, which means the anaemia condition or the
affect by the blood filtering , the different RBC group occur.
PLT histogram
 Description of RBC histogram
Hematology analyzer analyze the PLT in the range of 2 ～
30fl. The normal histogram curve is distribute
symmetry in the range of 2 ～ 20fl，the main apex is in
the 7.6 ～ 13.2fl. When the apex move left means that
the PLT size become small, the apex move right side
means that the size become biger.
If the curve area of the PLT become narrow means the
number decrease.
When small RBC and particulate of RBC exist, the
another apex will be on right side of main apex.
 Abnormal histogram of PLT

When the double apex occur on the distribution curve,

means that the PLT clot or number value is too low.
 RT-7600 HGB test principle

HGB calculate method

Lambert-Beer law
HGB=calibration coefficient ×Log10（Blank
value/sample value）
Clinic operation knowledge and
parameters introduction
 Blood sample collection
• 1. Anti-coagulating whole blood , Venous blood can be collected by
using vacuum negative pressure tube or ordinary method under at
atmospheric pressure, anticoagulant must be dropped to all venous
blood collection containers in advance, usually EDTA.K2.2H2O is
adopted as anticoagulant, with content of 1.5-2.2mg/ml.
• After collecting the blood, Prior to test, blood sample must be
thoroughly shaken, the recommended method is: shake up and down,
rotate test tube 3-5min, do not shake too violently.
• 2. Anti-peripheral blood collection, use the bullet blood tube for the
blood collection, the anticoagulant should be added, aspirate the 9.8ul
for testing.
• 3.Predilute peripheral blood collection: the typical collection method is
to pierce through finger end. Blood tube uses 20μL constant volume
blood tube or bullet blood tube. It is recommended to collect no less
than 30μL blood to facilitate double check. Please aspirate the 20ul
sample then add 700uL dilute liquid, form the1:36 liquid. The
instrument aspirate 300uL for test.
 Peripheral blood and Venous blood

Peripheral blood ：— convenient in collecting, low cost

——Test one time，easy clot
——Affect by the manual
—— Dilute sample need to stay for 3~5 minutes，
test within 20 minutes
Venous blood : —— Can test many times with sample
——High cost, need to add anticoagulant, not easy
to collect sample
——limit to some patients
——Good test accuracy (need to stay for some time),
Good test result within 8 hours.
 Introduction of reagent
• 1．Lyse reagent function：1.lyse RBC quickly, release HGB, ensure the
fragment of RBC has no effect on the WBC counting; 2.It help the HGB to
form stable compound for testing；3. Break down the membrane of the WBC,
release the plasm and ensure the integrity of the WBC.
• 2.Diluent function: 1.dilute the cell, prevent cell from clotting, let cell suspend
in liquid freely; 2. it has proper ion pressure, has the stable pervasion pressure,
which keep the blood corpuscle integrity and original volume; 3.It is actual one
kind of electrolyte with conductance feature, which has enough conductance
rate for the test.
• 3.Cleaning liquid function：mainly for the cleaning the ruby and parts of the
tubes; most of condition, it use the diluent for cleaning; there is much foam
when using cleaning liquid, after finish counting, sixty test (adjustable) finish,
clean by manual and clean before power off, the clean liquid will be used for
clean relative parts; the sampling needle is cleaning by the diluent.
• Concentration cleaning liquid (E-Z cleaning liquid)：E-Z washing liquid has
good performance on emulsification and catalysis, it help for dismissing the
solid smudge, make the protein smudge to water-solubility aminophenole, keep
the liquid path clean.
 Parameter introduction

20
P
A
R
A
M
T
E
R
 Test parameters

Full name Abbreviation (reference value) Unit

White blood corpuscle WBC（4～10， adult） 109/L

Red blood corpuscle RBC（4.0～5.5，adult） 1012/L

Hemoglobin HGB（110～160，adult） g/L

Platelet PLT （100～300） 109/L

 Parameter from histogram
Full name Abbreviation (reference Unit
value)
Lymphocyte percentage LYM%（20%～40%） %
Intermediate cells percentage MID%（3.5%～14%） %
Granulocyte percentage GRAN%（50%～70%） %
Mean corpuscular volume MCV （80～98fl） fL
Red cell distribution width RDW-SD（40～53） fL
standard deviation
Red cell volume distribution RDW-CV（0～15%） %
width CV
Mean platelet volume MPV（8.6～12.6） fL
Platelet distribution width PDW（9.8～17.2） fL
Platelet-large cell ratio P-LCR（16.9～46.7） %
 Parameter from the calculate

Full name Abbreviation (reference Unit

value)
Lymphocyte count LYM#（0.8～4.0） 109/L
Intermediate cell count MID#（0.14～1.2） 109/L
Granulocyte GRAN#（2.0～7.0） 109/L
Hematocrit HCT,（40-50%） %
Mean corpuscular MCH（26～32） pg
hemoglobing
Mean corpuscular MCHC（320～360） g/L
hemoglobin
concentration
Plateletcrit PCT（0.11%~0.28%） %
Parameter for calculate
Parameter for calculate
Instrument installation and test
 Unpack the instrument

Unpack the package and take off the material on the

carton
Take out the instrument from the bag carefully

Check the items according to the packing list

Keep the packing material well for future use

Environment requirement

No direct sunlight;
No massive dust or powder;
No strong electromagnetic radiation;
Sufficiently large flat and solid desktop.
 Power supply requirement

A.C.110V～220V

50/60Hz

96VA
 Instrument structure

• ① Display screen: Display the software

interface
• ② Indicator light: yellow at startup, turn
red when starting sample test, turn yellow
after test. 2

• ③ Feed key: built-in printer feeds paper

outward 3
• ④ Aspiration key: in sample analysis, 1 4
press this key, the instrument will aspirate
sample
• ⑤ Printer cover: print paperinstalling
position for built-in printer
• ⑥ Sampling needle: use sampling needle
to aspirate sample into counting cell 6
• ⑦ Aspiration key: in sample analysis,
press this key, the instrument will aspirate
sample, function the same as key ④ 5 7
 Rear view
• ⑴ Rear cover switch: turn this button,
open the rear cover to replace reagent
• ⑵ Keyboard interface: PS/2 keyboard 2
interface
• ⑶ Mouse interface: PS/2 mouse
interface 3
• ⑷ USB port 4
• ⑸ RS-232 serial port: to connect with 5
data receiving devices
• ⑹ Network interface 6
• ⑺ Power interface: to connect with 7
external power supply 1
• ⑻ Power switch: switch instrument 8
power
9
• ⑼ Diluent sensor
• ⑽ Grounding hole: used for instrument 10
grounding
• ⑾ Cleaner sensor 11

• ⑿ Cleaner port 12

• ⒀ Diluent port
14
• ⒁ Waste port 13
 Preparation of power on

Before startup, the operator shall check by

following steps to make sure system is ready:
• Check if the diluent, hemolytic agent and rinsing
solution meet the need of current test. Check if
the waste bottle is full, tubing system is ok and
without folded, all the connection is ok.
• Whether instrument power adapter is safely and
stably connected to power socket.
• Check whether built-in or external printer paper
is sufficiently installed in right position.
 Power on
Switch on the RT-7600, indicator light is on,
system begins to check component status,
transfer parameter needed by front end, run
blank tests, as shown in figure below:
 Blank test acceptable range

Parameter Range
WBC ≤ 0.2 × 109 / L
RBC ≤ 0.02 × 1012/ L
HGB ≤1g/L
HCT ≤ 0.5 %
PLT ≤ 10 × 109 / L

If blank test result exceeds this range, operator has to repeat above test steps until
test result is acceptable. If after 5 runs, test result still cannot meet specification,
please check reagent and tubing connection, and use “Backflush” function in
“Maintenance” module to eliminate block hole.
After blank test, one can fill out ID manually in Data Edit window, or click “Return”
button to return to main screen, go to Sample Test window again to run sample
test.
 Main screen
After the blank test finish the following figure showing

Click “Exit” turn back to main screen, showing on right figure

 System parameters’ setting
Enter “system setting” set the parameter
 Test
On the main manual click ”Test”，enter the sample test
window

Note:1、Click “profile ”change the sample number

2、If select the “real time” on system setting, after finish testing, it
will print the result automatically.
 Calibration
On the main screen click “calibrate” ，“calibrate by manual” or “calibrate
automatically”。

Calibrate by manual Calibrate automatically

 Quality control
RT-7600 has three kind of controls，L-J、X-B and X-R
L-J control setting
X-B control setting
X-R control setting
 Print history data
On the main screen click “history data” ，open the history data
list.
 Power off
On the main screen click “Power off”，the instrument will tip the
dialogue :

Click “ok”, the instrument will turn off the system, then it display ”please
turn off the instrument “, please switch off the instrument.
7600 liquid path structure
 Liquid path cleaning

The clean liquid flow into the pipeline from the clean bottle, 3 valve switch on, liquid
pass through three direction valve reach the WBC adjunctive pool and RBC
adjunctive pool, then 5 value open, the waste liquid goes into the waste chamber
under the vacuum negative pressure.
 Waste drain

The piston is pulled upon, negative pressure in the waste

chamber, the valve 10 and 11 are open, the liquid in RBC and
WBC flow into waste chamber; Valve 10 and 11 are close, the
piston is pulled down, the valve 4 is open, the waste in the
waste chamber will flow into waste bottle.
 Diluent's circuit

The piston of vacuum pump are pulled down, the diluent flow through valve 9(1-2) enter
the diluent pump; the piston of the vacuum are pulled up, the diluent flow through valve 9(2-
3), valve(1-2), valve(1-2), the diluent will go to cleaning chamber to washing the needle.
 Routine maintenance
• 1. Startup and shutdown process
At startup, the instrument will run mechanical component test and blank test,
for user to discover problem as soon as possible. While running shutdown
operation, daily shutdown maintenance program will be automatically
performed. After shutting off instrument power, just clean workbench and
instrument surface.
• 2. Automatic rinse
If number of samples instrument tests has reached the number preset by user,
then the instrument will run automatic rinse program. User can adjust
automatic rinse frequency at will. Refer to Chapter 7, System Setup. Rinse can
also be done in “Maintenance” menu.
• 3. Clean instrument surface
Keep instrument working environment clean.
Instrument surface can be cleaned with neutral detergent and wet cloth.
 Maintenance program 1
1 Backflush
Back flush gem hole, eliminate block-hole.
2. Drain counting cell
Drain off diluent in WBC and RBC counting cells.
3. Drain fluid circuit
Drain off liquid in fluid circuit.
4. Eliminate block-hole
The instrument will execute one special program for
rinse the tube.
 Maintenance program 2
5 Reagent filling
• In sample test, reagents will be automatically filled, if
drain operation is run or reagents are replaced, reagent
filling is needed.
• All reagents: fill diluent, lysis solution and cleaner
into related tubings.
Diluent: fill diluent into related tubing.
Lysis solution: fill lysis solution into related tubing.
Cleaner: fill cleaner into related tubing.
6. Rinse
Rinse probe, sampling needle and test tubing.
 Maintenance program 3
7 E-Z solution soaking
Suction 1.6ml probe rinsing solution with sampling needle to
fluid circuit and counting cells. The user shall perform E-Z
solution soaking operation once a week. Firstly, insert sampling
needle into E-Z solution bottle, then press “E-Z solution soaking”
key, run E-Z solution soaking operation.

8 Probe rinsing solution soaking

Suction 1.6ml probe rinsing solution with sampling needle to soak fluid
circuit and counting cells. The user shall perform probe rinsing solution
soaking operation once a week. Firstly, insert sampling needle into probe
rinsing solution bottle, then press “Probe rinsing solution soaking” key, run
probe rinsing solution soaking operation.
 Trouble shooting 1
1. Instrument cannot be started up
——check if instrument is powered on
• ——check if power plug gets loose or falls off
• ——check voltage

2. Instrument cuts power automatically

• ——check instrument power is connected or not
• ——check power cable is loose or not
• ——power off instrument internal circuit breaker, and restart.
 Trouble shooting 2
3. Temperature abnormal
——in software main menu, click “System Info” -> “System Status”, check
environment temperature, if not within 15C~35C range, restore instrument
ambient temperature to this range

4. Blank counting value higher repeatedly

——reagent is used up or not
• ——reagent is deteriorated or contaminated
• ——calibrate instrument
• ——check temperature or pressure if it is norma
 Trouble shooting 3
5 Parameter test incorrect
• ——calibrate instrument
• ——check if sampling needle location is right
• ——check if there is bubble in fluid syringe, piston slides
smoothly. If there is bubble, please make sure that reagent tubing
connection is normal.
• ——check solenoid valves if they work normally
• 6 Printer cannot print
• ——check printer if there is no paper
• ——check if normally connected
• ——check printer setup in system setup
 Trouble shooting 4
7 QC not in target range

• ——check reagent validity period

• ——check setup if it is right, and necessary to modify parameter.
• ——make sure that QC process is not contaminated
• ——test again in other method
The end

Thanks!