CONTENTS

1 INTRODUCTION, INTENDED USE 3

2 DESCRIPTION OF THE ANALYZER 4
2.1 Parts of the analyzer 4
2.1.1 Front panel 5
2.1.2 Rear panel 6
2.2 Fluidic Schematics 7
2.3 Function of fluidics 7
2.4 Control panels 7
2.4.1 Sampling bar 7
2.4.2 Status LED 7
2.4.3 Display 7
2.4.4 Keyboard 8
3 DESCRIPTION OF THE SOFTWARE 9
3.1 General 9
3.2 Menu system 9
3.3 Menu structure 10
4 OPERATING PRINCIPLES 11
4.1 Impedance method 11
4.2 Principle of the HGB Measurement 11
4.3 Parameters 12
4.4 Absolute and Linearity Ranges of Measured Parameters 13
5 ANALYZER INSTALLATION PROCEDURES 14
5.1 Unpacking 14
5.2 Connecting reagent containers 14
5.3 Connecting power cord 14
5.4 Connecting an external printer 14
5.5 Connecting the instrument to a host computer 14
5.6 Turning the instrument ON 15
5.7 Display contrast setting 15
5.8 Reagent priming 15
5.9 Turning the instrument OFF 15
5.10 Emergency handling 16
6 SOFTWARE SETTINGS 17
6.1 Setting Date and Time 17
6.2 Setting the name of the Laboratory 17
6.3 Results units setting 18
6.4 Setting reference limits 18
6.5 Fluid sensors 20
6.6 Printer settings 21
6.6.1 Troubleshooting guide for printing problems 23
6.7 User settings 24
6.7.1 Multi-user mode 25
7 ROUTINE UTILIZATION 26
7.1 Measuring process 26
7.1.1 Sampling 26
 7.1.2 Blank measurement 26
7.1.3 Calibration 27
7.1.4 Sample Analysis 30
7.1.5 Prediluted mode 32
7.1.6 Lyse boost 33
7.2 Quality control 33
7.2.1 QC database 34
7.3 Auto stand-by 35
7.4 Printouts 35
7.5 Patient results database management 37
7.6 Data Back up 39
7.7 Maintenance 40
7.7.1 Cleaning 40
7.7.2 Priming 40
7.7.3 Draining 40
7.7.4 Manual cleaning of the aperture 40
7.7.5 Weekly maintenance 41
7.8 Diagnostics 42
7.8.1 Device Statistics 42
7.8.2 Self test 42
7.9 Software update 43
8 APPENDIX "A": REAGENT SOLUTIONS AND CONSUMPTION 44
9 APPENDIX "B": TECHNICAL SPECIFICATIONS 46
10 APPENDIX "C": FLUIDIC Schematics 47
11 APPENDIX "D": ACCESSORIES 48
12 APPENDIX “E”: FINDING THE CORRECT LYSE VOLUME FOR VETERINARY APPLICATIONS 49

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1 INTRODUCTION, INTENDED USE
Humacount PLUS hematology analyzer is a fully automated cell counter for in vitro diagnostic use. The
compact instrument was developed for small to medium size labs with today’s technology.
The analyzer can up to process 60 samples per hour with the specified accuracy and reproducibility and has a
10,000 samples storage capacity including histograms.
It is intended to determine the following 18 hematology parameters from a 25 μl whole blood sample:

 WBC - LYM# - MID# - GRA# - LYM% - MID% - GRA% (three-part WBC differential)
 HGB - RBC - HCT - MCV - RDW - MCH - MCHC
 PLT - MPV - PCT - PDW
The instrument has a built-in veterinary software module, which allows measuring and determining 18
hematology parameters (three-part WBC differential) from different animal blood samples.
The interfaces allow sending results to an external printer (parallel port), to a host computer (serial port). By
using the integrated 3½” floppy disk drive, the instrument can be easily upgraded at any time and it can backup
data to floppy disk.

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2 DESCRIPTION OF THE ANALYZER
2.1 Parts of the analyzer
Humacount PLUS Hematology Analyzer consists of three main parts:
 Fluidic System: performs aspirating, sampling, diluting, mixing, lysing and rinsing, and additionally
generates the regulated measuring vacuum.
 Data Processing System: counts and measures blood parameters, generates and stores results
and histograms.
 Control Panels: include display, keyboard, parallel (external printer), and serial (computer) interface.

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2.1.1 Front panel

1 2 3 4 5 7 10 9 8

Figure 1. Front view

1. Graphic liquid crystal display (LCD)
2. Function buttons
3. Numerical keypad
4. Delete key
5. Cursor moving buttons
6. OK button
7. HELP key
8. Glowing LED
9. Aspirating needle
10. OK button

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2.1.2 Rear panel

Figure 2. Rear view

1. Reagent tubing connections
2. Waste full sensor connector
3. External keyboard connector
4. Serial (RS 232) connector for computer
5. Printer connector for external printer
6. Opening for air-flow rotor
7. Small power switch for separating the instrument from the mains power
8. 115/230 V selector
9. On/Off switch

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2.2 Fluidic Schematics
See Appendix “C”.

2.3 Function of fluidics

Sample flow prior to counting:
25 µl (50 µl in Prediluted mode) of anti-coagulated whole blood sample is aspirated by the sampling needle, and
4 ml of diluent is added into the mixing chamber. From here, 25 µl of primary dilution is aspirated and then added
with 5 ml of diluent into the RBC chamber (RBC dilution). The primary dilution remaining in the mixing chamber
flows via the tubing into the WBC chamber where the necessary (preset) amount of lysing reagent is added
(WBC dilution).

Dilution rates:
1. Primary dilution 1:160
2. RBC dilution 1:32 000
3. WBC dilution 1:196

Measurement times:
1. WBC counting: approx. 5 sec (float in xVM moves downward)
2. HGB measurement (float in xVM is at bottom position)
3. RBC/PLT counting: approx. 8 sec (float in xVM moves upward)

2.4 Control panels

2.4.1 Sampling bar
A push on the sampling bar triggers a measurement process.

2.4.2 Status LED

The status LED is located above the sampling bar (START button)
It has three colors indicating the actual status of the Fluidic System:

Color Analyzer status

 Green The analyzer is ready to work. New measurement can be initiated.

 Red The analyzer is performing a measurement process, no new measurement can be started.

 Yellow The analyzer is carrying out a maintenance process, or it is in stand-by.

2.4.3 Display
The display is a 240 x 128 dots, high contrast, CCFL backlit graphic LCD (Liquid Crystal Display) module.

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2.4.4 Keyboard
Keyboard is a 24-button foil keypad, including:
 Numerical buttons: for entering numerical data, and selecting menu items;
 Function buttons: for starting the action displayed with an icon above the function key at the bottom of
the LCD;
 Cursor moving arrows:  and , for moving between database items;
 and , for moving between parameter columns or menu levels;
 OK button: used for initiating a measurement cycle or confirming data;
 Del button: for deleting missed characters;
 Help button: for HELP function.

2.4.4.1 Function key description

Function key Action triggered
Exit from menu or action
Leave data-entry menu without saving any changes made to it
Confirm the results or changes made
Display histograms of the highlighted patient ID or QC Lot No.
Redo action (e. g. Blank measurement)
Modify sample/patient data
Send results to host computer
Print data (results, patient ID, QC)
Select between result pages (switch to PLT)
Select between result pages (switch to RBC, WBC)
PAGE-DOWN key in a multi-page menu
PAGE-UP key in a multi-page menu

Change the Levey-Jennings chart scaling (16 or 64 days)

Patient type selection

Patient type selection in veterinary mode (dog/cat/horse/other)

Confirm error
Go to local menu (database, measurement)

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3 DESCRIPTION OF THE SOFTWARE
3.1 General
The integrated software controls instrument operations, displays, stores, recalls data, and allows the User to
perform quality control (QC) and calibration procedures.
Pressing the OK button validates newly entered data, or activates/enters the highlighted item.

3.2 Menu system

The instrument uses a menu system to initiate actions, and to modify settings. In the menu system there are
three ways to navigate between menus and menu items.

 Position the selection bar to the desired item with the and buttons and press the OK button. This will
validate the highlighted menu item. Press the function key to return to the previous menu level. This
method is suggested while learning instrument operation.
 Pressing the number that corresponds to the desired menu item will also validate it. In this case there is no
need to press OK. Pressing the 0 (zero) button has the same effect as the function key. This method
is much more efficient in practice.
 You can move between the higher and lower menu levels using the  and  keys.

The existence of a sub-menu is indicated by a ► symbol at the end of the menu line.

Some results can be displayed in table format. In this case, button 3 (three) has the same effect as the Page Up
key, and button 9 (nine) has the same effect as the Page Down key. Use these buttons to move between table
pages. Further positioning keys during browsing the database are 1 (Jump to top) and 7 (Jump to bottom).

Some menu items have selected and deselected (on and off) states. These items are marked with a  symbol in
front of the item number. The filled  mark indicates selected state. Selecting the menu item toggles its state.
Other menu items work as so-called “radio-buttons”. It means that only one of the buttons of the group can be on
at once. In some cases, more buttons are displayed together, though they correspond to different groups. In this
case, a horizontal line separates the groups.

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3.3 Menu structure
Measure Measure Local Prediluted mode Cleaning
Database Measure blank Maintenance Priming
Maintenance Patient type Patient type Draining
Calibration Repeat last sample
Quality control Limits
Settings
Shut down View database Local Go to specified record Select by date time, and ID
Backup database Selection Select all
Restore and view Change sort order Deselect all
Send selected records***
Backup selected records Unsorted
Delete selected records Sort by time
Sort by sample ID
Back up one day Sort by patient ID
Back up selected
Cleaning
Priming Prime all
Draining Prime diluent
Diagnostics Device information Prime lyse
Service Statistics Prime cleaner
Self test
Drain all chambers
Calibrate Calibrate MCV and MPV Drain RBC chamber
View calibrations Calibrate HCT and PCT Drain WBC chamber
Calibration settings Factorial calibration Drain MIX chamber
Calibration with one measure
Calibration with two measures
Calibration with three measures
Calibrate prediluted mode

Set QC reference QC level 1 Set date and time

QC measure QC level 2 Day.Month.Year
View table of QC measures QC level 3 Month.Day.Year
View QC diagram QC level 4 Year.Month.Day
Set QC level QC level 5
QC level 6 Human
Male
Date and time Female Cat
Laboratory Baby Dog
Units Toddler Horse
Limits Child Other1
Fluid sensors Diluent Veterinary limits ** Other2
Printer settings Lyse Other3
User settings Waste Other4
Other5
Shut down Single-user mode
Logout * Multi-user mode
Prepare for shipment Add new user *

* only in Multi-user mode

** only if veterinary mode is available
*** only if serial communication is enabled

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4 OPERATING PRINCIPLES
4.1 Impedance method

The impedance method counts and sizes cells by detecting and measuring changes in electrical impedance
when a particle in a conductive liquid passes through a small aperture.

Each cell, passing through the aperture – where a constant DC current flows between the external and internal
electrodes – causes some change in the impedance of the conductive blood cell suspension. These changes are
recorded as increases in the voltage between the electrodes.
The number of pulses correlates to the number of particles (CBC - Counting Blood Cell). The intensity of each
pulse is proportional to the volume of that particle (WBC, RBC, PLT histograms). Pulses are counted only in
channels (in terms of femtoliters, fl) which are between the pre-set lower and upper discriminators.

4.2 Principle of the HGB Measurement

The lysed 1:196 sample dilution can be measured by a cyanmethemoglobin method. The reagent lyses the red
blood cells, which release hemoglobin.
Hemoglobin iron is converted from the ferrous (Fe 2+) to the ferric (Fe3+) state to form methemoglobin, which
combines with potassium cyanide (KCN) to produce the stable cyanmethemoglobin, or hemoglobincyanide.
Subsequently, the HGB concentration is measured photometrically.
Note:The above-mentioned measuring method is used to determine the HGB concentration. The HGB
concentration can be measured using cyan-free lysing reagents as well. In this case the effect is
the same but the used lyse is environmental-friendly reagent.

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4.3 Parameters
White Blood Cell – WBC Number of leukocytes.
(cells/l, cells/µl) WBC = WBCcal x (cells/l, cells/µl)
Red Blood Cell – RBC Number of erythrocytes.
(cells/l, cells/µl) RBC = RBCcal x (cells/l, cells/µl)
Hemoglobin concentration - HGB Measured photometrically at 540 nm; in each cycle blank
measurement is performed on diluent
(g/dl, g/l, mmol/l) HGB = HGBcal x (HGBmeasured – HGBblank)
Mean Corpuscular Volume - MCV Average volume of individual erythrocytes derived from the RBC
(fl) histogram.

Hematocrit – HCT Calculated from the RBC and MCV values.

HCTpercentage = RBC x MCV x 100,
(percentage, absolute) HCTabsolute = RBC x MCV
Mean Corpuscular Hemoglobin – MCH Average hemoglobin content of erythrocytes, calculated from RBC
and HGB values.
(pg, fmol) MCH = HGB / RBC
Mean Corpuscular Hemoglobin Calculated from the HGB and HCT values.
Concentration – MCHC MCHC = HGB / HCTabsolute
Unit of measurement is displayed according to the one chosen for
(g/dl, g/l, mmol/l) HGB result (g/dl, g/l or mmol/l)
Red cell Distribut. Width - RDW-SD The distribution width of the erythrocyte or platelet population
And derived from the histogram at 20% of peak
Platelet Distribut. Width - PDW-SD
(fl)

Red cell Distribut. Width - RDW-CV

And
Platelet Distribut. Width - PDW-CV xDW-SD = RDWcal x (P2 - P1) (fl),
(absolute) xDW-CV = RDWcal x 0.56 x (P2 - P1) / (P2 + P1)
by the factor of 0.56 CV is corrected to the 60% cut
Platelet – PLT Number of thrombocytes (platelets)
(cells/l, cells/µl) PLT = PLTcal x (cells/l, cells/µl)
Mean Platelet Volume – MPV Average volume of individual platelets derived from the PLT
(fl) histogram.

Thrombocrit – PCT Calculated from the PLT and MPV values.

PCTpercentage = PLT x MPV x 100,
(percentage, absolute) PCTabsolute = PLT x MPV
White blood cell differential: Absolute values counted in the channels determined by the three
 LYM, LY% : lymphocytes WBC discriminators:

 MID, MID% : monocytes and some 1. 2. 3.

eosinophil granulocytes 1.: RBC-LYM discriminator
LYM MID GRA 2.: LYM-MID discriminator
 GRA, GR% : neutrophil, eosinophil 3.: MID-GRA discriminator
and basophil granulocytes

Percentages calculated from the absolute WBC value.

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4.4 Absolute and Linearity Ranges of Measured Parameters
In the linearity range, the instrument works within the specified accuracy. Out of this linearity range, the system
is able to display the results, but without the specified terms.
If the value is over the maximum range, the instrument can not measure it and the result is marked with an E
(Error) flag. In this case, pre-dilution of the sample is recommended (e.g. very high WBC). For the Prediluted
mode see section 7.1.5 of this manual.

The linearity ranges of primary parameters in normal measuring mode:

Parameter Linearity Ranges Maximum Unit
WBC 0...75 100 109 cells/liter
RBC 0...15 20 1012 cells/liter
PLT 0...700 1000 109 cells/liter
HGB 0...250 400 g/l
HCT 0...100 - %
MCV 30...150 - fl
MPV 3...30 - fl

The linearity ranges for 1:3 predilution mode:

Parameter Linearity Ranges Maximum Unit
WBC 4...300 400 10 cells/liter
9

RBC 2...40 60 1012 cells/liter

PLT 200...3000 4000 109 cells/liter

User Manual HumaCount Plus 13/51

5 ANALYZER INSTALLATION PROCEDURES
5.1 Unpacking
Only trained Service Personnel should perform unpacking. It's recommended to carefully inspect for any damage
or missing items at the time of unpacking.
CAUTION! Before the first operation, let the instrument warm up to room temperature for at least 6 hours;
otherwise the temperature gradient could cause water condensation, which may damage the electronic parts.

5.2 Connecting reagent containers

Waste
Sensor

All containers should be left open to provide free airflow.

5.3 Connecting power cord

Connect the power cord to the connector located on the rear panel and then to a grounded electrical outlet.

Do not, for any reason, power the instrument from an electrical outlet that is not properly
grounded.

5.4 Connecting an external printer

Connect external printer to the parallel port located on the rear panel. A built-in parallel interface protocol is
provided for Epson 9-pin or compatible, Epson 24-pin or compatible, and for HP PCL4 compatible printers. (For
printer settings see 6.6.)

5.5 Connecting the instrument to a host computer

The instrument has a built-in serial communication protocol that allows connecting the analyzer to a host
computer. This serial interface is able to send results (including histograms) to the host computer. The serial I/O
settings are in the service menu system. Therefore if you want to connect your instrument to a host computer,
please call your Service Person.

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5.6 Turning the instrument ON
1. Turn on the connected external printer
2. Turn on the power supply of the instrument by flipping the small power switch to I position.
3. Press the large switch located on the left side of the rear panel to power up the instrument.

An automatic start-up procedure will be initiated. During start-up, the following screen is displayed. The software
version number appears in the bottom line when the program is started.

When the start-up procedure is completed, the MAIN MENU is

Main menu displayed.
1.Measure ► Sub-menus are indicated with an ► arrow in the end of the line.
2.Database ► In this case, you can use  key to enter the selected sub-menu.
3.Maintenance ►
4.Calibration ►
5.Quality control ►
6.Settings ►
7.Exit ►
CAUTION! Wait 5 minutes before starting any
measuring process. This time is required to reach
the optimal working temperature.

5.7 Display contrast setting

From the Main menu go to Maintenance menu. After selecting Service submenu, you can adjust the display
contrast (visibiltiy / brightness). Up arrow () lightens, down arrow () darkens the screen.

5.8 Reagent priming

In some cases, a priming cycle must be performed. The instrument will perform the cycle automatically if the fluid
sensors are on, and the instrument detects that there is not enough liquid in the tubing system. Priming cycle
should be run in the following cases:
 At installation
 In case of extended time out of use
 After any component replacement related to the Fluidic System

5.9 Turning the instrument OFF

EXIT (7)
Exit
(SHUT DOWN (1))
1. Shut down The analyzer should not be switched off by simply unplugging it.
2. Preparing for shipment For shutdown, always use EXIT menu (7), item 1.

The software will prompt you for confirmation. Press 1, and the
analyzer will perform a priming cycle to fill the chamber to avoid
dust and salt build-up, and shuts itself down.

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The second item should be used when the instrument is to be shipped. In this case, the instrument will ask you
to connect the cleaning tube kit with 100ml of distilled water. Follow the instructions appearing on the display.
EXIT (7)
! Message 5001/19300 PREPARING FOR SHIPMENT (2)
Remove reagent tubing at rear reagent inputs Here, the instrument asks you to remove tubing connectors, so
(Diluent, Lyse and Cleaner) that they can be drained. Leave waste connector on.

Next, you should

connect at least 100 ml of distilled water, ! Message 5002/19300 which the
instrument uses to wash out any reagents Connect min 100 ml distilled water to reagent remaining in the
system. Waste must still be on. inputs using cleaning tube kit.

! Message 5003/19300 Next, the analyzer asks you to remove cleaning tube kit (waste
Remove cleaning tube kit. Keep reagent inputs must still be on).
free. When finished, the analyzer will ask you to power off the system.

5.10 Emergency handling

In case of emergency situation - like catching fire the instrument (short-circuit, etc.) - cut off the current
immediately and use the fire-extinguisher.

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6 SOFTWARE SETTINGS
Settings The SETTINGS sub-menu allows you to set customer-defined
limits, hardware and software configurations required for proper
1.Date and time ►
instrument operation.
2.Laboratory
3.Units
From this sub-menu, you can access the date and time, limits,
4.Limits ►
patient type, and other settings related to the instrument.
5.Fluid sensors ►
6.Printer settings
7.User settings ►

6.1 Setting Date and Time

The date and time of each measurement is stored with the results. This menu allows setting the built-in clock of
the instrument and date format.
SETTINGS (6)
Date and time
DATE AND TIME (1)
1.Set date and time
○2.Day.Month.Year By selecting item 1, You will enter date and time setting mode
○3.Month.Day.Year (next screen).
●4.Year.Month.Day
Items 2-4 are radio-buttons, so only one can be selected at once.

SETTINGS (6)
Set date and time DATE AND TIME (1)
SET DATE AND TIME (1)
The current date and time are
Enter the actual date and time using the numerical keypad. The
12.05.2000 15:00 form of the date is as it was set in the previous menu.

Confirm by pressing the button.

6.2 Setting the name of the Laboratory

This menu allows entering laboratory information, which will be printed in the header of reports generated by the
instrument.

Set laboratory name SETTINGS (1)

The name of the laboratory is: LABORATORY (2)
Hematology-1 A maximum of 40 characters can be entered in each line of 4 (or
8* if Extended header enabled by Service) either using an
external standard PC (US-layout) keyboard connected to the
This name will be printed in the header instrument, or by scrolling each letter character using the and
of the printouts buttons.
Move between character places using and  keys.

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6.3 Results units setting
SETTINGS (6)
Units UNITS (3)
Units can be set in a sequential order. The arrow on the right
Count unit cells/l ↓ indicates that there are more options for the entry. Select among
HGB unit g/l them using the and buttons. When done, pressing OK opens
PCT, HCT unit % the next unit type for setting. When the last entry is set,
RDW, PDW mode sd confirms data, and returns to the Settings menu.

The possible units for above parameters are as follows:

Count unit cells/liter, cells/μl

HGB unit grams/liter (g/l), grams/deciliter (g/dl), millimols/liter (mmol/l)
PCT, HCT unit percentage (%), absolute (abs)
RDW, PDW mode Standard Deviation (sd), Coefficient of Variation (cv)

6.4 Setting reference limits

This function allows the User to specify the patient reference (so-called normal) ranges used in your laboratory.
Lower and upper limits - stored in the memory - are displayed and can be modified according to the patient type
selected in the PATIENT TYPE menu.
SETTINGS (6)
Limits LIMITS (4)
1.Human In this menu, you can set the limits of the different patient types.
2.Male
3.Female
4.Baby
5.Toddler
6.Child
7.Veterinary limits ►

The next three pictures show the reference range settings of Female patient type.
SETTINGS (6)
Patient limits LIMITS (4)
Female FEMALE (3)
Lyse 0.70 ml
WBC 5.0 - 10.0 10^9/l Modify the highlighted lower and upper limits with the numerical
RBC 4.00 - 5.00 10^12/l keypad. Confirm with OK.
HGB 120 - 160 g/l By pressing and function key buttons, you can turn to
HCT 36.0 - 48.0 % other pages with additional parameters.
MCV 76 - 96 fl
MCH 27.0 - 32.0 pg

Patient limits If 0.0 - 0.0 is

specified as lower and upper limit, MCHC 300 - 350 g/l the parameter
will not be checked for its proper PLT 150 - 400 10^9/l range.
PCT 0.0 - 0.0 %
MPV 8.0 - 15.0 fl
PDWs 0.0 - 0.0 fl
PDWc 0.0 - 0.0 %
RDWs 20.0 - 42.0 fl
RDWc 0.0 - 0.0 %

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 Patient limits
LYM 1.3 - 4.0 10^9/l
MID 0.2 - 0.7 10^9/l
GRA 2.5 - 7.5 10^9/l
LY% 25.0 - 40.0 %
MI% 3.0 - 7.0 %
GR% 50.0 - 75.0 %

Actual lyse quantity can be modified in Measure mode before sampling in 0.1 ml steps to a maximum extent of
0.2 ml for that particular measurement (see 7.1.6).
In the same submenu the software also allows the user to set the different veterinary normal ranges and the
quantity of the added lyse reagent.

Limits SETTINGS (6)

1.Dog LIMITS (4)
2.Cat VETERINARY LIMITS (7)
3.Horse Other1…5 submenus let the user to set any kind of animal type,
4.Other1 name, limits and lyse quantity. The next three figures show the
5.Other2 reference ranges settings of veterinary patient type.
6.Other3
7.Other4
8.Other5

Patient limits SETTINGS (6)

Dog LIMITS (4)
Lyse 0.60 ml DOG (1)
WBC 6.0 - 17.0 10^9/l
Modify the highlighted lower and upper limits using the numerical
RBC 5.50 - 8.50 10^12/l
keypad. Confirm with OK.
HGB 120 - 180 g/l
By pressing and function keys, you can move to
HCT 37.0 - 55.0 %
other pages with additional parameters.
MCV 60 - 77 fl
MCH 19.5 - 24.5 pg

If 0.0 - 0.0 is specified as lower and upper limit, the

Patient limits
parameter will not be checked for its proper range.
MCHC 310 - 340 g/l
PLT 200 - 500 10^9/l
PCT 0.0 - 0.0 %
MPV 0.0 - 0.0 fl
PDWs 0.0 - 0.0 fl
PDWc 0.0 - 0.0 %
RDWs 0.0 - 0.0 fl
RDWc 0.0 - 0.0 %

Patient limits Actual lyse quantity can be modified in Measure mode before
LYM 1.0 - 4.8 10^9/l sampling in 0.1 ml steps to a maximum extent of 0.2 ml for that
MID 3.0 - 12.0 10^9/l particular measurement (see 7.1.6).
GRA 0.1 - 1.3 10^9/l
LY% 12.0 - 20.0 %
MI% 3.0 - 10.0 %
GR% 62.0 - 87.0 %

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6.5 Fluid sensors
Fluid sensors check for presence of the most important reagents: diluent and lyse.
If both waste connectors are used, the waste container full alert, called waste sensor is operating.

Cleaner reagent has no sensor, operator should check cleaner amount daily.

If any of the fluid sensors get out of order, the analyzer can still continue operating, if the malfunctioning sensor
is manually disabled.
SETTINGS (6)
Fluid sensors FLUID SENSORS (5)
□ 1. Diluent
■ 2. Lyse
■ 3. Waste Any sensor can be switched on and off by toggling its state using
the OK button.

Here, Diluent sensor is disabled.

If any of the fluid sensors is switched off, an S warning can be

seen in the top-left corner of the screen during
measurement.

WASTE HANDLING – VERY IMPORTANT

Waste contains poisonous substances (because of possible cyanide content) and human origin substances
causing biohazard. These substances are representing potential danger to environment. For this reason, safe
handling of the waste liquid is very important.

The following steps should be made for environmental protection and safety reasons:

Neutralization of biohazard effect

 Independently of the fact that the waste contains cyanide or not, you should make this step.
 Put 2 ml/l hypochlorite solution into the waste. Close the cap, shake the container and wait 1 hour.
 Annihilate the waste by spilling it into the drain system.

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6.6 Printer settings
The instrument supports the following printers or printer modes:
 HP DeskJet
 HP LaserJet
 Epson 9-pin
 Epson 24-pin
 Epson 24-pin in 9-pin mode
 Canon BJC
 Seiko DPU414
Any printer being able to emulate any of the above modes can be connected to the instrument.
To set up the instrument for your printer, go to the “Settings/Printer Settings” menu. Choose one of the
possibilities using the up and down arrow keys within the text fields, and fill in the numerical fields using the
numerical keys.

The general characteristics of the printable area of printer paper are below:

The paper is defined by its size:

it can be standard (A4, Letter) or any other
custom size paper (actual size).

Printers cannot print on the whole surface of the

paper. The blank area is described by the
physical Margins, which may vary by printer
models. The paper area inside the physical
margins is called printable area.

Top margin and Left margin settings are used

for determining the exact place where the
results inside the printable area are printed.

If more than one result is to be printed on one

sheet of paper, use Vertical spacing to
determine the distance between the reports.

Printer settings On the first page of Printer Settings, printer type, graphics mode
Printer: and paper settings can be set.
Seiko DPU414
Mode: Select the Printer matching your printer hardware.
Normal Wide ↑
Physical margin: Normal Different Modes result in different printout sizes. Choose the one
Paper: A4 matching your needs. Options are Small, Normal, Normal Wide.
Unit: inch Recommended mode is Normal.
Size 8.27 x 11.69
It is suggested to make a sample printout after setting up printer
settings.

Physical Margin is used to correct the printout for a printer having lower printable area. Bigger margin means
bigger physical margin. Alter the ‘normal’ setting only, if the right side of the printed result is missing, or appears
in the next line.
Select Paper, or enter correct Size parameters: either in inches or in cm, specified at Unit.

User Manual HumaCount Plus 21/51

The second page of printer settings enables changing further margins, vertical spacing of reports, and other
options.

If rollpaper is used instead of sheets, set Rollpaper to Yes.

Printer settings
Margins: If all results are to be printed automatically after each
Left: 0.00 Top: 0.00 measurement, set Autoprint to Yes.
Vertical spacing: 0.50
Rollpaper: No Printout format can be specified as one of the following:
Autoprint: No  Full with histograms
Printout format: Full with histograms  Text only
Table format: Normal table format
 Short, text only
One result per page: No

If you set One result per page to Yes, each measurement will be printed on another page.
Printer settings The third page of printer settings holds the options for the
Print ranges: Yes individual result-printning.
Print flags: Yes
Print warning flags: No If Print Ranges is set to Yes, the upper and lower limits (if
Print measure time: specified in Settings/Limits) will be printed next to each measured
In case of warning parameter for verification.

If Print Flags is set to Yes, and any measured value is out of the normal range, or there was any error, those
will be shown on the printout as well. It is recommended to set it to Yes.

If Print warning flags is enabled, any flags that was shown on the result screen will be included with the
printout.

The Print measurement time option has several settings.

The time of measurement gives information about the counting process, and is a good verification of the results.
If it was too high, it means that the measurement should not be accepted. Possible settings are: “No”, “Always”,
“In case of warning”.
Recommended setting is “In case of warning”!

22/51 User Manual HumaCount Plus

6.6.1 Troubleshooting guide for printing problems

Problem Possible reason, remedy

Printer does not respond,  Printer is off. Turn it on.
no printout.  Printer is not connected to the analyzer.
Connect it to the parallel port at rear.
 Printer is not On-line. Switch it to On-line mode.
 Printer is out of paper. Feed the printer with paper.
Strange signs or letters appear on  The selected printer type does not match your printer. Select the
the printout. appropriate type in Printer Settings menu.
 Printer is not set up properly for HP or Epson
(or compatible) mode. Modify the printer setup. Consult the printer’s
manual
Right side of the printed report is  Decrease Margin settings in the Printer Settings menu.
missing, or appears in the next  Try selecting another Mode making a smaller printout.
line.
The printed report is too small,  Try selecting another Mode resulting in larger printout.
there is a lot of space on the
paper.
The end of the printout appears on  Enter the correct Paper size.
the next page.  Try increasing Margin.
It seems that one more patient  Enter the correct Paper size.
report could fit on the same page.  Try decreasing Margin, Top margin, Vertical spacing.
The printed result is not centered  Modify Left margin.
horizontally.
The printed result is not centered  Modify Top margin.
vertically.
The distance between two results  Modify Vertical spacing.
is too small or too big.
After printing, the printer does not  It is quite usual for bubble-jet or laser printers.
eject the paper. Do not repeat printing.
But if you repeat printing, it When the page is full, or you leave the actual menu, the printer will
appears in more copies. eject the paper automatically.

User Manual HumaCount Plus 23/51

6.7 User settings
User settings SETTINGS (6)
●1.Single user mode USER SETTINGS (7)
○2.Multi user mode This menu allows configuring the instrument for single-user or
multi-user mode.

Care must be taken when changing to multi-user mode.

SETTINGS (6)
USER SETTINGS (7)
User settings MULTI USER MODE (2)
○1.Single user mode By selecting item 2, the instrument enters multi-user mode, and a
●2.Multi user mode new item appears on the screen: Add new user.
3.Add new user

Multi-user mode allows more people using the same instrument

with individual personal settings, so-called profiles.

In a multi-user environment users have their own profiles describing their rights within the system. In an
environment like this there must be a so-called supervisor having the right to add, delete users and modify their
rights.

The software of the instrument has three levels of user-rights: Basic, Advanced and Supervisor.

A Basic-level user has absolutely minimal rights within the system. These are: performing measurements and
entering patient data before the measurement.

A user of the Advanced level has the power to modify settings of the instrument: as described in chapters 6.1-
6.6, do QC and Calibration. An Advanced user can also modify patient data when browsing in the database.
A Supervisor has the ability to do all the above, and additionally to modify user rights and user passwords.

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In Multi-user mode, there must always be a user with Supervisor rights, otherwise only a Service-person can
reset the instrument.

SETTINGS (6)
User information
USER SETTINGS (7)
User ID 1 ADD NEW USER (3)
Name
Frank Smith____________________ To each new user, the software assigns an individual ID.
Level Basic
Active Yes In the next field, a user name of 32 characters can be specified.
Password

When the name is entered, the level should be defined. The default setting for Active is Yes. Use this option if
you want to disable a user.

The last data to enter is the password, a maximum of 8 alphanumeric characters.

6.7.1 Multi-user mode

Login If the analyzer is in multi-user mode, a logon screen will be

User ID 0________ displayed during start-up, asking for a user ID and a password.
Password
User name

Login When the User ID is entered (confirm with OK), the user name
User ID 1 corresponding to the ID appears in the lower line. If the user
Password ****_____ name was correct, enter the password, and confirm with the
User name function key. If the password was correct, the analyzer
Frank Smith continues initialization and is ready to work.

In multi-user mode the Exit menu is changed: a Log-out menu point appears.

EXIT (7)
Exit
1. Shut down When the user logged in has finished working with the instrument,
2. Preparing for shipment it is not necessary to switch off. From the main menu, selecting
3. Logout EXIT (7) brings up the EXIT menu.

LOGOUT should be used instead of shutdown, as in this case the

log-in screen will be displayed waiting for the next user to log in.

User Manual HumaCount Plus 25/51

7 ROUTINE UTILIZATION
7.1 Measuring process
7.1.1 Sampling
The aspirating tip (a needle with rounded end) is used to draw sample from the tube containing the blood to be
analyzed. It can not always be seen, as it has two positions:

a) Retracted position: inside the instrument (the status LED is either red, yellow or off).
b) Aspirating position: in front of the sampling bar; it can be activated only in certain menus associated directly
with the sampling process; status LED is green.

1. Invert the closed sample tube 11 times to achieve a homogenous sample. Do not Shake!
2. Take off the cap of the sample tube.
3. Immerse the aspiration needle well into the sample.
4. Push the sampling bar or the START/OK button.

The instrument draws 25μl of sample, and the aspirating needle is retracted while its outer surface is
automatically rinsed with diluent. This ensures a low carry-over between samples. Simultaneously, the status
LED turns red.

During aspiration, hold the sampling tube in a stable position (for approx. 1 second) until you see the status LED
flashing and hear the beeps indicating the end of the sampling process.

After these signals, you can remove and recap the sampling tube.

Attention! If you hurt yourself during analysis biohazard substances can cause infection! Use always
rubber gloves!

Make sure to immerse the sampling needle well into the sample, otherwise it can cause
erroneous sampling, giving inaccurate results.
Important! When the needle stops for a while during its movement upwards, the needle must
be out of the sample, otherwise the analyzer makes another aspiration in this state. This
extra aspiration can also cause an inaccurate measurement.

7.1.2 Blank measurement

Blank measurement is used for checking the cleanliness of the system and reagents.
Blank measurement must be performed in the following cases:
 Once daily, before the sample analyses (this is done automatically before the first measurement in
MEASURE).
 After any reagent change (it can be activated manually from the MEASURE/ MEASURE BLANK menu).
 After any component replacement, which is closely related to a measuring process (aspiration, dilution,
counting, rinsing).
User must accept the blank values by pressing .

Blank measurement can be repeated by pressing .

If any of the tested parameters has High blank value, the

message: Unsuccessful blank measure appears at the top
of the screen.

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There are 3 regions for blank value handling:
1. Optimal, all results are OK.
2. Blank is high, * flag is displayed at related parameter results.
3. Blank is too high, no result displayed in measurement mode.

Blank measurement ranges:

Parameter 1. No flag at parameter 2. * flag at result 3. E (error) flag at result

HGB 0-10 g/l 10-25 g/l > 25 g/l
WBC 0 - 0.5 x10 cells/µl
3
0.5-1.0 x10 cells /µl
3
> 1.0 x103 cells /µl
PLT 0 - 25 x103 cells /µl 25 - 50 x103 cells /µl > 50 x103 cells /µl
RBC 0-0.05 x106 cells/µl 0.05-0.5 x106 cells /µl > 0.5 x106 cells /µl

Accepted blank values are essential for proper calibration and QC measurement. For this reason, no calibration
or QC measurement can be performed without accepted blank values.

Quality control (QC) measurement and Calibration can be performed only if all blank values are in the 1st region.

7.1.3 Calibration
Calibration is the procedure used to standardize the instrument by applying the necessary correction factors.

It is recommended to do calibration in the following cases:

1. At analyzer installation, before beginning the analyses.
2. After replacing any component, related to the process of dilution or measurement.
3. When quality control measurements show any systematic error (bias) or they are outside predefined
limits.
4. At regular time intervals (determined by the lab itself).
5. If you want to use the instrument in Prediluted mode.

Calibration can be performed in two ways:

1. User can enter calibration factors - without any calibration measurements – using the numerical keypad.
2. One-, two- or three-fold measurements of control or special calibrations material with known parameters.
In this case, the instrument automatically calculates new factors using the following formula:

Assigned value x Stored factor

New factor =
Measured value(s) (or average of those)

CAUTION! New calibration will invalidate the previous one. Old values cannot be retrieved,
but can be reviewed in the VIEW CALIBRATIONS menu.
Calibration can be initiated by choosing Calibration in the Main Menu.

Calibration CALIBRATION (4)

Item 1 initiates calibration procedure.
1.Calibrate
2.View calibrations
Item 2 displays previous Calibrations
3.Calibration settings
Item 3 opens calibration settings dialogue.

User Manual HumaCount Plus 27/51

7.1.3.1 Factorial calibration
If the CALIBRATION MODE has been previously set to Factorial Calibration, the factors can be set manually in
the 0.80 - 1.20 range.

Calibration CALIBRATION (4)

RBC 1.02 CALIBRATE (1) (factorial)
MCV 1.05 Set previously calculated factors using the numeric keys, confirm
RDWc 1.12 with OK.
PLT 1.00 Only parameters concerning the selected MCV, MPV / HCT, PCT
MPV 1.00 mode are displayed.
HGB 0.92
WBC 1.01

7.1.3.2 Automatic calibration by measurement

If the CALIBRATION MODE is set to one-, two- or three-fold measurements, the instrument will perform
calibration measurements with hematology blood control.

Calibration CALIBRATION (4)

RBC 4.57 10^12/l CALIBRATE (1) (automatic)
MCV 85 fl Set the target values of the control material using numeric keys.
RDWc 16.4 % Use OK to accept a value.
PLT 255 10^9/l
MPV 9.1 fl Specify 0 as target value for parameters should be omitted from
HGB 136 g/l calibration.
WBC 7.4 10^9/l
After setting the target values, press to validate and start
calibration measurements.

Target values for the calibrated parameters can be set in the following ranges:

Parameter Low limit High limit

RBC 1.00 8.00
HCT 0.1 0.6
MCV 50 120
RDW CV 10 50
PLT 30 800
PCT 0 2
MPV 5 15
PDW CV 5 50
HGB g/l 30 300
WBC 1.0 30.0
Having set the required target values, perform measurements on hematology blood control, hold the sample
tube in sampling position and press the sampling bar.
Press to accept results.

The actual and the total number of calibration measurements are

shown in the first line.

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 After calibration the new factors are displayed. The actual factors
Calibration are shown in parentheses for reference. Offered calibration
RBC 1.02 (1.03) factors can be accepted by pressing .
MCV 1.05 (1.01)
RDWc 1.12 (1.09) The factors are not modified, but get flagged:
PLT 1.00 (0.92) – if 0 was targeted (no calibration)
MPV 0.98 (0.96) B if blank was not in the 1st region
HGB 1.08 (1.05) E if factor is out of 0.80-1.20 range
WBC 1.15 (1.11)

7.1.3.3 View calibrations

From the Calibration menu, you can monitor previous calibrations. The instrument logs all calibration events, and
displays them in the following format.
CALIBRATION (4)
Date Time OpID RBC MCV RDWc PLT
VIEW CALIBRATIONS (2)
23.05.2004 10:15 0 1.03 1.07 0.85 1.04 Use ← and → keys to browse between parameter columns.
29.05.2004 11:21 0 1.01 1.07 0.86 1.01
Exit pressing function key with symbol.
02.06.2004 09:51 5184 1.00 1.00 1.00 1.00

7.1.3.4 Calibration settings

By choosing CALIBRATION SETTINGS, you get to the following screen:
CALIBRATION (4)
Calibration settings CALIBRATION SETTINGS (3)
● 1.Calibrate MCV and MPV Items 1-2 select between calibration target,
○ 2.Calibrate HCT and PCT Items 3-6 select between calibration modes.
● 3.Factorial calibration
Select item 7 to perform CALIBRATION FOR PREDILUTED
○ 4.Calibrate with one measurement
MODE. By choosing this point, you can calibrate the instrument
○ 5.Calibrate with two measurements
with the same processes (factorial, automatic) as in normal mode.
○ 6.Calibrate with three measurements
□ 7.Calibrate prediluted mode

When Calibrate prediluted mode is selected,

Calibration/Prediluted mode will appear at the first line on the Calibration screens.

The previous Prediluted calibration factors can not be monitored in the VIEW CALIBRATIONS submenu.

To perform factorial calibration, you should set target parameter (1 or 2), and choose item 3, Factorial
Calibration. In this case, User must have had the necessary amount of measurements with the control material,
based on which an average value can be calculated. This average value is used for fine-tuning the calibration
parameters.

User Manual HumaCount Plus 29/51

7.1.4 Sample Analysis
7.1.4.1 Sample preparation
Use K3-EDTA anti-coagulated fresh whole blood as sample. Prior to sampling, mix the sample gently by
inverting it 11 times. Do not shake as this could damage the blood cells.

7.1.4.2 Sample information

The instrument allows the User to enter sample information for each sample that has been, or will be measured.
An external PC keyboard (standard USA layout) must be connected to the instrument before turning it on.
Sample information can be entered in two ways:

 immediately before measurement, or

 in the Database menu

To enter sample information before sample analysis, press the function key. This brings up the following
screen:

Here sample ID and patient data can be specified (name, sex,

Sample information date of birth). Also, the name of the doctor having ordered the
Sample ID: 1 laboratory analysis can be given.
1
Date: 2004.05.12. Patient name will appear on the LCD below the PLT histogram
Patient ID: (second page), and in the header of the printed result sheet.
Name:
John Johnson
Birth. 1958.04.16. Male
Doctor: dr. Harrison

On this screen, the patient age can be entered in years or in months depending on your selection.
As the requirements vary from lab to lab and is strictly related to
Sample information
regular laboratory work, the user cannot change this option.
Sample ID: 1 Contact service for further information.
1
Date: 2004.05.12.
Patient ID:
Name:
John Johnson
Age: 25 years ↓ Male
Doctor: dr. Harrison

Enter the name using up to 32 alphanumeric characters, (“A-Z”, “0-9”, space, comma, dot and parentheses “()”).
Use arrow keys to move between characters, backspace to delete missed characters.
Press Enter after entering the name, cancel with Esc or , confirm with .

7.1.4.3 Results
At the end of a measuring procedure, the following screen is displayed, including all measured and calculated
parameters as well as the WBC, RBC and PLT histograms.

Results and histograms will be stored automatically in the memory,

without any operator confirmation. (First panel of two)
If normal ranges are set (not 0.0), parameters will be verified and
marked by:
+ if the value is over,
- if the value is under the range specified.

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 If there is any kind of error, or the blank measurement is too high, E error flag will appear at the erroneous
parameter and no results will be displayed (---).
 If there are warnings or errors, a  flag will appear preceding the actual result.

In the last line of the first result screen, warning flags can be displayed. If the last line is clear, it means that no
errors or warnings were found during measuring cycle. The meaning of each warning flag and the recommended
user action is included in the next table.

Flag Meaning Recommended user action

W WBC three part  Repeat the measurement. Possibly lyse problem.
warning or WBC three  Check the discriminators in the WBC histogram. If the discriminators are
part diff. unsuccessful in the proper place (the populations can be separated by eye) then the
results can be accepted.
E No WBC three part  Possibly lyse problem, but in some pathological samples (too high
lymphocytes), it can be happened.
H HGB blank is high, or  Repeat the blank measurement and accept it.
no HGB blank  Possibly lyse or diluent problem.
B WBC blank is high, or  Repeat the blank measurement and accept it.
no WBC blank  Possibly lyse or diluent problem.
L WBC/RBC limit  Check the 1. RBC-LYM discriminator. If it is in the minimum point (or
warning close to it), accept the results. Otherwise repeat the measurement.
 If the retried action makes very similar results and the discriminator is in a
wrong place then the MID and GRA results are OK, but the WBC and
LYM results can be higher because of the RBCs.
R Too many RBC cut  Repeat the measurement. Possibly lyse problem.
from WBC  If the WBC measuring time is too high (more than 8 sec.) it could be
aperture clogging. In that case perform cleaning and repeat the
measurement.
M* WBC coincidence is  The results are out of the linearity range. Make a dilution with an external
too high. Linearity dilutor with a pre-defined dilution range. Do not forget to correct the
error. results with the defined factor.
D WBC data package  Perform cleaning, redo measurement (aperture clogging).
errors  If it is a general problem, please call your Service Personnel.
S WBC time error  The same action as in case of the D warning flag.
C WBC clogging  Aperture clogging. The same action as in case of the D warning flag.
p PLT blank is high, or  Repeat the blank measurement and accept it.
no PLT blank  Diluent problem. Replace diluent, open a new tank.
b RBC blank is high, or  Repeat the blank measurement and accept it.
no RBC blank  Diluent problem. Replace diluent, open a new tank.
l RBC/PLT limit  The RBC/PLT valley is too high. It is rather a diagnostic flag. If the
warning discriminator is in a wrong place (in the PLT or RBC histogram) then
repeat the measurement for a correct PLT result.
k RBC peak warning  Perform cleaning and repeat the measurement (clogging).
 If it is a general problem, change the RBC aperture.
m* RBC/PLT coincidence  The same action as in case of the M warning flag.
is too high. Linearity
error.
d RBC/PLT data  The same action as in case of the D warning flag.
package errors
s RBC/PLT time error  The same action as in case of the D warning flag.
c RBC/PLT clogging  The same action as in case of the C warning flag.
*M or m warning flag means that the coincidence is too high so the results are out of the linearity range - see
Section 4.4. In that case, make a predilution with an external dilutor and use the Prediluted mode (see section
7.1.5).

User Manual HumaCount Plus 31/51

In the next figure, one of the long-format printout patient results with histograms is shown.
Humacount PLUS 07.03.2000 15:38 #2 Female
At the bottom of the screen, warning
flags are displayed and the WBC
results have the  flag, meaning that
the WBC three-part differential does
not seem to be correct.

In this patient result, the L warning

flag means that the RBC-LYM
discriminator is not sure and that is
why the W warning flag (WBC three
part warning) appears in the last line.

With a little diagnostic experience, the User can accept the results, because just a very small part of the RBC is
in the LYM channel. (The discriminator is in the minimum point of the dotted RBC line) Similarly, the MID
population seems to be correct, thus the results can be accepted.

Second panel of the result screen including the PLT distribution

curve and measuring times.

7.1.5 Prediluted mode

The software has a spezial Prediluted mode, useful in the following situations:
 If sample values are above the linearity range (see section 4.4)
 In case of very small sample volumes
 In case of capillary blood samples

In this Prediluted mode, you have to make an external 1:3 (or 1:10*) pre-dilution of whole blood, as in the
following example:
Predilution: 1 UNIT OF SAMPLE + 3 (or 10*) UNITS OF DILUENT !
Example: if 20µl capillary tubes are used for blood collection, one should add 60µl (or 200µl*) of pure
diluent to create a proper pre-dilution.

<MEASURE LOCAL MENU>

Measure local menu To select Prediluted mode, you must enable it in the Measure
1.Patient type ► local menu. Now, the instrument is ready to measure the
□2.Prediluted mode prediluted sample.
3.Maintenance ► 1:3 (or 1:10*) warning flag will appear on the WBC or PLT
4.Repeat last sample histogram of the result screen.
5.Limits ►
* The instrument is capable of measuring in two
pre-dilution modes 1:3 and 1:10. The user cannot
change these modes. Contact service for further
information.

The results are corrected with the predilution factor (1:3 or 1:10) automatically.
Prediluted mode has its own calibration factors. To calibrate the instrument for this mode,
see the Calibration settings section of this manual.

To disable Prediluted mode, you must re-enter Measure local menu and deselect its box.

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 <MEASURE LOCAL MENU>
Measure local menu
1. Cleaning MAINTENANCE (3)
2. Priming ►
3. Draining ► From the Measure local menu, you can also access some parts
of the maintenance menu. There you can make priming,
cleaning, or draining of the chambers.

7.1.6 Lyse boost

Would you find, that the preset lyse volume is not giving
satisfactory results for WBC 3-part differential, you can modify
the actually used lyse volume in ±0.1 ml steps to a maximum of
±0.2 ml, by pressing the up and down arrows on the keypad. A
small L+, L++ (+0.2ml), or L-, L--(-0.2ml) sign indicates this
change on the screen.
For further information, see Appendix E: Finding the correct
lyse volume for vet applications.

7.2 Quality control

By measuring control materials, day-to-day reproducibility can be monitored. In this submenu, both target value
and acceptable ranges for each parameter can be determined for the different QC levels.

Humacount PLUS provides six different Quality Control profiles, so-called levels. You can set up six individual
reference sheets for each control material (e.g. low, normal and high control blood). QC measurement results
will be added to the selected level, is indicated at the top right corner.
The target values of the control material should be set only once, at the beginning of the QC
measurements. Resetting parameters deletes previous QC results of the active level.

CAUTION! Any change in the QC material setting deletes previous QC results, therefore it is
strongly recommended to have them printed out before making any changes to parameters.
QUALITY CONTROL (6)
Quality control
This menu allows setting and monitoring data related to the QC
1.Set QC reference procedure.
2.QC measure
3.View table of QC measures Use commercially available control material. The preparation
4.View QC diagram and measurement process is the same as with patient samples.
5.Set QC level ►
Actual QC level: 1

Set QC level QUALITY CONTROL (6)

●1. QC Level 1 SET QC LEVEL (5)
○2. QC Level 2 Select the level you wish to use.
○3. QC Level 3 The active level is displayed in the top right corner on QC
○4. QC Level 4 related screens.
○5. QC Level 5
○6. QC Level 6

User Manual HumaCount Plus 33/51

QUALITY CONTROL (5)
SET QC REFERENCE (1)

QC reference values
QC Level: 1
LOT No.: __________________
Exp. date: 12.05.2004.
WBC 5.5 - 19.5 10^9/l
RBC 5.00 - 10.00 10^12/l
HGB 80 - 150 g/l
HCT 24.0 - 45.0 %
MCV 39 - 55 fl

Both target values and acceptable ranges can be specified. Only the parameters displayed on these screens can
be specified. Modify displayed values using the numerical keyboard. Pressing the OK button validates values. To
move on to further parameters, press the page down function key.

Quality control measurements can only be made after an optimal blank measurement result
has been accepted (all parameters were in the 1st range).

QUALITY CONTROL (5)

QC MEASURE (2)
After selecting target values (or targeted level),
select this menu to do a QC measurement. After
the measurement has been completed, the
following result screen will be displayed. It is
similar to the normal measurement, but in the first
line there is Quality Control as an ID.

CAUTION! A result will be accepted only if it is confirmed with the key.

7.2.1 QC database
The database of measured and stored QC results can be displayed at any time in table or graphic
(Levey-Jennings) formats. The QC measurement results will have subsequent ID numbers.
QUALITY CONTROL (5)
SmpID Date PatID VIEW TABLE OF QC MEASURES (3)
□3 2004.05.02 07:06
□4 2004.05.03 07:12 Move the selection bar over entries with and ↓ buttons.
□5 2004.05.04 07:02
□6 2004.05.05 07:36 Moving among parameters with  and  buttons.
□7 2004.05.08 06:56
□8 2004.05.09 07:01
□9 2004.05.10 07:02
□10 2004.05.11 0650

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 CAUTION! Any change in the QC material settings is followed by deletion of the QC
database, so it is strongly recommended to print them out before modification.

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The analyzer makes graphic figures of the primary parameters (WBC, RBC, HGB, HCT, MCV and PLT).
QUALITY CONTROL (5)
VIEW QC DIAGRAM (4)

Assigned values (target value and accepted range) as well as

averages, standard deviations (StDev) and coefficients of variation (CVar) are calculated based on the QC
measurements and are displayed together with Levey-Jennings charts. The dotted lines show the acceptable
range.

7.3 Auto stand-by

In measurement mode, the analyzer will automatically go to stand-by if it is left alone - without starting any new
measurement - for more than 15 minutes. While in stand-by, the aspirating tip is inside the analyzer and the
chamber is filled with diluent. When the sampling bar is pressed, the chamber will be cleaned, the tip comes out
and the analyzer is ready for the next measurement. The same process is performed when entering and leaving
measurement mode.

7.4 Printouts
When required, the following items can be sent to an external printer by pressing the function key button.

 Database result(s) (table format).

 Database (specified patient results with histograms).
 QC result (Levey-Jennings chart).
 QC result(s) (table format).
 Last measured blank result.
 Last measured patient result (with histograms).
 Last measured QC result.
 Device Information and Device Statistics.
 Self test result.
 Set parameters.
Different printout formats of typical patient-result printouts are shown in the next figures (the appropriate printout
format can be selected in SETTINGS/PRINTER SETTINGS submenu – see 6.6).

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Full printout format with histograms:

In this printout normal ranges (limits), flags and warning flags are all enabled. The printing modes of these
parameters can be selected within PRINTER SETTINGS submenu.
If normal ranges are set, flags are enabled, and patient values are over or below the limits specified, the result
out of the range is marked with an +/- mark.

Normal printout format without histograms (Text only):

Hematology-1
07.03.2004 15:38 #2 Female

This printout format is similar to the previous one, but it does not include
the histograms. The laboratory name has moved into the first line.

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7.5 Patient results database management
Patient results are stored in the memory in chronological order, and they can be retrieved at any time.
Memorizing capacity is 10,000 measurements including the complete parameter list, histograms, flags, sample
data, dates and times of the measurements.

DATABASE (2)
Database
1.View database From this menu monitoring of stored measurements can be
2.Backup database done. Select item 1 to view the database in table format.
3.Restore and view

DATABASE (2)
VIEW DATABASE (1)
SmpID Date PatID Pressing  or  button accesses the remaining, non-visible
?7 2004.05.02 14:56 20040502a12 parameter results. With  or  button you can scroll between
¦8 2004.05.04 14:59 HR0058AS4 the results one-by-one. Pressing the 3 and 9 buttons has the
? High05 2004.05.04 15:06 PWF55A
same effect as PageUp and PageDown scrolling keys.
? 08 2004.05.04 15:08 LNK23-B
? 12A7 2004.05.04 15:15 MK4FED
¦ UF6A8 2004.05.04 16:00 550462L
Select patient results by pressing the OK button. Its box will be
? HEM2 2004.05.04 17:26 HEMA0034A
filled.
¦ PR02 2004.05.04 17:29 ZZ3ZH

From the database table screen, WBC, RBC and PLT

histograms can be displayed by pressing . By pressing the key, the second panel with PLT results
can be accessed.

Step-by-step browsing among histograms of the selected tests is made by pressing LEFT and RIGHT ARROW
function key buttons.( and )

Database local menu <DATABASE LOCAL MENU>

From the database table screen, you can go to the Database
1.Go to specified record
2.Selection ► local menu by pressing the key.
3.Change sort order ►
4.Backup selected records
5.Delete selected records

<DATABASE LOCAL MENU>

GO TO SPECIFIED RECORD (1)
Go to specified record
Item 1 brings up a screen asking for Go to the first record measured after: parameters
defining any given sample (date, time ID) and 02.05.2000. 00:00 jumps to it. If
any ID is left as 0, it means searching only by and has Sample ID: date/time.

and has Patient ID:

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 <DATABASE LOCAL MENU>
Selection SELECTION (2)
1.Select by date, time and ID Besides checking samples one-by-one, selection can also be
2.Select all done within Database local menu.
3.Deselect all SELECT ALL checks all boxes, DESELECT ALL clears all
boxes.

<DATABASE LOCAL MENU>

Select specified data records SELECTION (2)
Select data records measured between: SELECT BY DATE, TIME AND ID (2)
02.05.2000. 00:00… 02.05.2000. 23:59
and has Sample ID in range: Select a range by date, time and/or ID number (see next screen)
Entering 0 as ID# means searching only by date. Corresponding
and has Patient ID: results will be marked with a filled box.

Change sort order <DATABASE

●1. Unsorted LOCAL
MENU> ○2. Sort by time
CHANGE SORT ORDER (3) ○3. Sort by Sample ID
○4. Sort by Patient ID
The way results are displayed can be selected within this
menu.

If set, samples appear according to the rule selected.

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7.6 Data Back up
This submenu allows backing up stored data to a floppy disk, and monitoring database stored on floppy disk.
Before menu selection, insert a 3.5” floppy disk into the drive located on the left-bottom panel of the analyzer. A
floppy disk can hold 800 samples.
Back up database DATABASE (2)
1.Back up one day BACKUP DATABASE (2)
2.Back up selected records BACKUP SELECTED will save entries that have been selected
in the database view (either one-by-one or by range selection).

DATABASE (2)
BACKUP DATABASE (2)
Backup data BACKUP ONE DAY (1)
Specify the day whose records you wish to backup to a floppy
Day to back up: disk.
2000.05.24.

When you have

selected the day or data to be saved, and ! Message 5104/12210 confirmed it
with the key, the instrument will prompt 35 data record(s) will be saved on 1 disk(s). you for an
empty disk. Insert an empty floppy disk!

Possible error messages related to backing up data:

The instrument gives this warning in case you tried to save data
! Message 5106/12210 of a day, which had no data, or no data are selected.
No such type of data.

This warning appears, if the disk you inserted had errors on it, or
STOP! Error 1300/12210
was write-protected.
Cannot write archive data! Check disk!
It may be unformatted, full, or bad! Check the write protection of the disk or if necessary, insert a
Do you want to retry? new, formatted disk.

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7.7 Maintenance
From this submenu, the User can initiate maintenance procedures such as cleaning, priming, draining and
preparing the analyzer for shipment.
MAINTENANCE (3)
Maintenance
1. Cleaning Select the required submenu.
2. Priming ►
3. Draining ►
4. Diagnostics ►
5. Service

7.7.1 Cleaning
Select item 1 (CLEANING) in the above MAINTENANCE menu.

This action is recommended in case any clogging problem is experienced (C error flag).

This is a relatively strong decontamination process, cleaning out possible dirt or clog from the system using high
voltage electronic pulses, cleaner solution and high-pressure backflush.

7.7.2 Priming
MAINTENANCE (3)
Priming PRIMING (2)
1. Prime all
2. Prime diluent During the priming cycle, the fluidics system gets rinsed with a
3. Prime lyse relatively large amount of diluent. It differs from the process in a
4. Prime cleaner start-up procedure; as in the latter case a simple filling up of the
fluidics is performed.

If fluid sensors are on, then the analyzer makes these procedures automatically, otherwise the User must initiate
them activating the appropriate item within this submenu.

7.7.3 Draining
MAINTENANCE (3)
Draining DRAIN (3)
1. Drain all chambers
2. Drain RBC chamber Draining is used to drain the required chamber, or chambers
3. Drain WBC chamber before servicing (e. g. manual cleaning, dismounting of the
4. Drain MIX chamber aperture). Drain all will also drain the volume limiter and the
ground electrodes as well as each chambers.

7.7.4 Manual cleaning of the aperture

The risk of clogging of the aperture by protein build-up is held at a minimum level in several ways. These are as
follows:

 The specific cleaning cycle can be done from MAINTENANCE/CLEANING submenu.

 Aspiration of the specific cleaner as patient sample.
 Applying high-voltage electric impulses to the aperture.
(Done after each measurement.)

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If manual cleaning of the aperture is required, instructions listed below should be followed:

1. Drain the chamber: MAINTENANCE/DRAIN

2. Open the side door (right side).
3. Remove the reference electrode connection and the U-shaped metal fixing.
4. Remove the aperture assembly from the measuring chamber.
5. Pull out the measuring tube from the part containing the electrode.
6. Cleaning of the aperture can be performed by placing it into a 10% hypochlorite solution, or into an
ultrasonic cleaning bath for approximately 1 minute.
7. Rinse well with distilled water.
8. Put back the measuring tube to the electrode holding part, and the U-shaped metal fixing.
9. Install the aperture in the measuring chamber, and connect the reference electrode.
10. Execute a priming cycle.
11. Measure blank until you reach acceptable blank results.

7.7.5 Weekly maintenance

User should do weekly maintenance on the first workday, before starting up the analyzer.

7.7.5.1 Cleaning washing head

There could be salt build-up in the lower surface of the washing head, which
may cause malfunction during operation. User should clean the lower surface
of the aspirating tip cleaning head using a soft cloth immersed in warm tap
water to remove salt build-up.

See steps 1 and 2 below:

1. Exit Measurement menu and wait until needle stops, then open side door.
2. To clean, gently rub the lower surface of the washing head, then close the side door.

7.7.5.2 Peristaltic pump maintenance

The pump installed in the instrument is maintenance free. However, would
you experience leakage from the pump, or vacuum error, you may replace
the tube used. To do it:
1. Open the side door.
2. Disconnect the tubes of the pump connector, and turn the lock at the end
with the connectors. Replace the tube with the replacement tube provided
with the instrument.
3. Put the cassette back and reconnect the tubes. Make sure they are
connected to the right connector (top: to waste output connector, bottom:
tube from valves)

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7.8 Diagnostics
The DIAGNOSTICS submenu provides important information about the analyzer, statistics and the built-in Self
test.
Diagnostics
MAINTENANCE (3)
DIAGNOSTICS (5)
1. Device information
2. Statistics Select the desired item.
3. Self test

MAINTENANCE (3)
DIAGNOSTICS (5)
Device information DEVICE INFORMATION (1)
Model: Humacount PLUS Here, device-specific information can be retrieved. Model name,
Serial No.: 1541 serial number, software version and the date of the compilation
Version: 2.3 of the software.
Compiled: Dec 22. 2000

7.8.1 Device Statistics

MAINTENANCE (3) Device statistics

DIAGNOSTICS (5) Measurements 6283
STATISTICS (2) WBC clogging 4
This menu includes important information RBC clogging 3 about the
measurements: the total number of Vacuum error 1 measurements,
clogging, vacuum and the other error Comm error 0 numbers.
MHori error 2
MVert error 0
Mdilu error 0

7.8.2 Self test

Self test is a procedure to check for proper operation of essential components of the instrument.
Self test should be performed in the following cases:

 Installation.
 After replacing any component.
 In case of extended time out of use.

The automatic Self test procedure can be initiated from the MAINTENANCE menu. During the test, you can see
its progress. The components, that have passed the test, get checked.

After completion of the Self test procedure, results will be displayed.

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 The first panel of result screen includes the date of testing and
Self test results Device Information.
Date of testing: 17.11.2000
Model: Humacount Plus
Serial No: 1541
Version: 2.3
Compiled: Dec 22. 2000.
PCPNIF Firmware version ---
MPU Firmware version: 2.0

The second and third panels of the result screen include every
Self test results test result. At the end of the result line, OK, HIGH, LOW, or
HGB dark: 275 OK ERROR is displayed, which means that the actual test result is at
HGB light 17723 OK the normal range (OK), higher (HIGH), lower (LOW) than the pre-
Electr. Voltage: 51.2 V OK determined limits, or the result is an error (ERROR).
current: 879 uA OK
offset -1.9 mV OK
Ampl. test: 20000 pls OK
peak: 1641 mV OK
dev: 53 mV OK

Self test results On the third panel of the result screen, the overall test result is
Noise test: 1 pls/5 sec OK displayed which can be Successful (in case of every test result is
Atm 978 mBar OK OK), or Errors found (if HIGH, LOW or ERROR results appear).
Vacuum: 218 mBar OK
drift: 3 mBar/10sec OK

Overall result: Successful

7.9 Software update

The User, following the steps below can easily update the instrument software:

1. Shut down the instrument, and switch it off.

2. Insert the upgrade floppy disk in the drive.
3. Switch on the instrument and follow the on-screen instructions.
4. When instructed, remove the disk from the drive.

At the next start-up, the instrument will run the new software.

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8 APPENDIX "A": REAGENT SOLUTIONS AND CONSUMPTION
Content
1. [DIL]: HC-Diluent Cat.- No.: 17400/10 (20 litres)
Diluent, isotonic saline solution used to dilute whole blood specimens and to rinse the
fluidic system between measuring procedures.
Buffer < 0.05 %, pH 6.9  0.2
Sodium Sulphate < 1.20 %
Detergent < 0.01 %
Stabiliser < 0.30 %

2. [LYSE]: HC-Lyse CF Cat.- No.: 17400/20 (1 litre)

Cyanid free lyse reagent
Used to prepare blood hemolysate for WBC and HGB measurement.
Sodium Chloride < 0.15 %, pH 7.4  0.5
Organic Acid < 0.25 %
Quarternary ammonium salts < 7.30 %
Preservative < 0.01 %

3. [CLEAN]: HC-Cleaner Cat.- No.: 17400/30 (1 litre)

Cleaning solution
Used to perform cleaning process of the fluidics.
Sodium Chloride < 0.10 %, pH 8.0  0.2
Potassium Chloride < 0.03 %
Proteolytic Enzymes < 0.02 %
Detergent < 0.40 %
Stabiliser < 0.30 %

Stability
The reagents are stable up to the expiry date on the label when stored at 15...35°C protected from light in a dark
place.
Once opened and installed on the instrument the reagents are stable for 60 days at 15...35°C. Instability due to
contamination is mostly indicated by cloudiness or colour change. This reagent should be replaced at once.

DO NOT FREEZE! Replace reagents once frozen.

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Precautions and Notes
 Leave the reagents at room temperature for at least 24 hours before use.
 HUMAN reagents have been optimised for the HUMACOUNT system. Use of and mix-up with third party
reagent have not been validated and may cause erroneous results.
 [LYSE] contains quaternary ammonium salts which may cause irreversible effects to skin and eyes in case
of contact. Wear appropriate safety equipment (gloves and lab coat). In case of contact rinse immediately
with plenty of water. If swallowed, induce vomiting and seek medical advice.
 [DIL] and [CLEAN] contain no hazardous substances in reportable quantities. In case of contact of skin or
eye rinse with water.
 Specimens, controls and material coming into contact with them should be handled as potentially infectious
and must be disposed off in accordance with the applicable local regulations in force.

Average reagent consumption of Humacount PLUS:

Number of daily Diluent Lyse Cleaner

measurements ml/test ml/test ml/test
1 287 16 33
2 160 8,5 17
5 85 3,9 7,1
10 59 2,4 3,8
20 47 1,7 2,2
50 39 1,2 1,2
100 36 1,1 0,9

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9 APPENDIX "B": TECHNICAL SPECIFICATIONS
Sample volume: 25 µl whole blood (50 µl in Prediluted mode)
Aperture diameters: 80 µm (RBC/PLT), 100 µm (WBC)
Throughput: approx. 55 tests/hour

Characteristics: Accuracy Reproducibility Carry over Test range

WBC 3% < 3% < 1% 4.0 – 20.0
RBC 2% < 2% <1% 4.00 – 5.00
HCT 2% < 2% <1% 30.0 – 50.0
MCV 2% < 1% N/A 70 – 100
HGB 2% < 2% <1% 100 – 160
PLT 4% < 4% <3% 200 – 400

Fault statistics: RBC clogging< 1% of measurements (normal use)

WBC clogging < 1% of measurements (normal use)
Clog prevention: High-voltage burn for both apertures in each measurement cycle
Cleaning procedure: High-voltage burst on the aperture, high-pressure backflush and chemical cleaning of
aperture using Cleaner reagent
Automatic cleaning: To prevent protein and lipid build-up in WBC chamber, programmable between 0
(none) – 50 measurement cycles, factory set to 20
Quality control: 6 QC levels (profiles) including: average, ± range, SD and CV for all measured and
calculated parameters, 16- and 64-day Levey-Jennings charts, separate QC
database
Calibration: Automatic (by calibrator) with 1, 2 or 3 measurements, or factorial (manual)
calibration of WBC, HGB, RBC, PLT, MCV, RDW, MPV, PDW
Automatic and factorial calibration of Prediluted mode
Monitoring of calibration factors by calibration events (not available for Prediluted
mode)
On-board CPU: Pentium™ PC
Data storing capacity: 10,000 results incl. histograms (16Mbytes DiskOnChip module)
Interfaces: RS 232 (computer) in 150, 300, 600, 1200, 2400, 3600, 4800, 9600, 14400, 19200, 38400,
57600, 115200 baud rate (8 data bit, no parity, 1 stop bit)
Parallel output (printer) in HP PCL4, Epson 9-pin, Epson 24-pin, Canon BJC, Seiko DPU-
414 or compatible mode
3.5” floppy disk drive
PC standard external keyboard, USA layout
Display: 240x128 dots graphic, backlit LCD
Keyboard: 24 buttons foil, START bar
Dimensions: 370x470x380 mm
Net weight: 17 kg
Power requirements: 110/230 V; 50/60Hz; 230VA
Installation category: II. (OverVoltage)
Primary mains fuse: T5A/250V
Protect class: I.
Operating temperature: 10 - 35 C

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10APPENDIX "C": FLUIDIC Schematics

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11APPENDIX "D": ACCESSORIES
This appendix includes the contents of Humacount PLUS pack:

 Humacount PLUS Hematology Analyzer.

 Humacount PLUS User's Manual.
 Humacount PLUS Reagent Tubes (marked with colours) :
- Diluent tube (green)
- Lyse tube (yellow)
- Clean tube (blue)
- Waste tube (red)

 Humacount PLUS Cleaning Tube Kit: Diluent

Lyse

Clean

 Humacount PLUS Caps for reagent containers

(Marked with the same colors as the Reagent Tubes).
 Humacount PLUS Power cable.
 Humacount PLUS Spare Part: Pump tube

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12APPENDIX “E”: FINDING THE CORRECT LYSE VOLUME FOR
VETERINARY APPLICATIONS
The software running on the instrument uses some default lyse volumes, pre-set at software installation.
However, the required lyse volume depends on several parameters. Lyse volume can be adjusted in the
SETTINGS/LIMITS menu for each species. An additional option to modify lyse quantity by  0.2 ml is available
during analysis. (see 7.1.6.)

The manufacturer can not guarantee that pre-set volumes will always give the best results. It is therefore
strongly recommended that the user oneself shall find the necessary volumes for the veterinary application. This
requires some experiments, but the result validated a user doing vet practice will be worth it.

The two most important parameters influencing lysing are lysing time and lyse quantity. You can not change the
lysing time, as it is fine-tuned for the lysing reagent supplied by Diatron.
Above you could read that lyse of quantity can be adjusted either at patient limits, or right before measurement.

In the following figures, you can see examples of over- or under-lysed samples.

Press  to increase the lyse quantity (+0.1 ml) if the separation between lysed RBCs and WBC populations is
poorly differentiated, resulting in increased WBC and LYM counts.

Under-lysed sample, some RBCs counted The same sample with lyse boost (+0.1 ml)
as WBCs WBC =13.7, good 3-part differential
WBC = 16.9, LYM% high

You can see that a lower amount of lyse will result in a less differentiated curve. If you increase the lyse quantity
it will be more differentiated, and well evaluated.

Press  to decrease the lyse amount (-0.1 ml) if the WBC histogram seems to be shrunk to the left, i.e. the
different WBC populations are overlapped. This can inhibit proper separation of WBC populations.

Over-lysed sample, LYM and GRA overlapped The same sample with lyse decreased (– 0.1 ml)
WBC = 20.6, poor 3-part differential WBC = 21.0, good 3-part differential

An over lysed sample will have all its cells shifted down towards the smaller cells. It is because of too much
lysing reagent shrinking all cells to the size of their nuclei.

With lyse quantity decreased, the cells will not shrink that much, and can be differentiated easily.

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 Revision history:

Revision Section Modification By At

Icons used in SW

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