logos biosystems Luna II User manual

Catalog #: L10001
Document #: LBSM-MD-ML-LUB-001
Version: VL1712-01

2
www.logosbio.com
IMPORTANT
LUNA™ Automated Cell Counter is a Laboratory Electrical Instrument for
Scientific Research Use Only, BUT NOT A MEDICAL or THERAPEUTIC or IN
VITRO DIAGNOTICS DEVICE.
Table of Contents
Safety Information -----------------------------------------------------------------------------------------------------
4
General Guidelines ---------------------------------------------------------------------------------------------------
6
Environment Conditions ---------------------------------------------------------------------------------------------
7
Chapter 1 –Introduction ------------------------------------------------------------------------------
8
1.1 Product Overview ---------------------------------------------------------------------------------------------
8
1.2 Product Contents ---------------------------------------------------------------------------------------------
10
1.3 Product Specifications ---------------------------------------------------------------------------------------
11
1.4 Product Description -------------------------------------------------------------------------------------------
12
Chapter 2 –Setting up ----------------------------------------------------------------------------------
15
2.1 Installation -------------------------------------------------------------------------------------------------------
15
2.2 Start-Up Screen -----------------------------------------------------------------------------------------------
16
2.3 Setting -----------------------------------------------------------------------------------------------------------
17
2.4 Setting the Date and Time ----------------------------------------------------------------------------------
18
2.5 Save Options ---------------------------------------------------------------------------------------------------
18
2.6 Counting Options ----------------------------------------------------------------------------------------------
20
2.7 Result/Printer/Calculator Options -------------------------------------------------------------------------
23
Chapter 3 –Counting Cells --------------------------------------------------------------------------
24
3.1 Sample Preparation ------------------------------------------------------------------------------------------
24
3.2 Loading Samples into Slides -------------------------------------------------------------------------------
24
3.3 Counting Cells -------------------------------------------------------------------------------------------------
25
3.4 Using the “Tag” Function ------------------------------------------------------------------------------------
27
3.5 Cell Size and Number Distribution ------------------------------------------------------------------------
28
3.6 Using the Dilution Calculator -------------------------------------------------------------------------------
33
3.7 Saving the Image and Generating a Report ------------------------------------------------------------
35
3.8 Printing the Counting Results ------------------------------------------------------------------------------
38

3
www.logosbio.com
Chapter 4 –Reanalyzing Saved Count Results --------------------------------------
41
4.1 Importing Previous Counts ----------------------------------------------------------------------------------
41
4.2 Recounting ------------------------------------------------------------------------------------------------------
41
4.3 “Previous Count” Button -------------------------------------------------------------------------------------
42
4.4 Setting up the Protocol for Counting ---------------------------------------------------------------------
44
Chapter 5 –Maintenance and Troubleshooting -------------------------------------
47
5.1 Cleaning ---------------------------------------------------------------------------------------------------------
47
5.2 Changing the Battery -----------------------------------------------------------------------------------------
48
5.3 Calibrating the Counter --------------------------------------------------------------------------------------
49
5.4 Updating the Firmware ---------------------------------------------------------------------------------------
53
5.5 Troubleshooting ------------------------------------------------------------------------------------------------
54
Chapter 6 - Ordering Information ---------------------------------------------------------------
56
Chapter 7 –Purchaser Notification -----------------------------------------------------
57

4
www.logosbio.com
Safety Information
For best results, users of the LUNA™ Automated Cell Counter must follow the instructions below in
addition to the general precautions for using electrical instruments.
1. Users must be careful to avoid electric shock while operating the instrument. Do not touch it and
other components with wet hands. Do not place it in a humid environment such as an incubator.
For operating environment, see page 7.
2. Trypan Blue Stain is a known hazardous material. While handling the solution, always wear
gloves to avoid exposure.
3. Before use, make sure that the input voltage is compatible with the instrument’s power supply
voltage.
4. For optimal operation, place the instrument on a flat bench and avoid any vibration.
5. Turn on the instrument only after connecting both ends of its power cord to the wall outlet as well
as the instrument. Always turn off the instrument before disconnecting the power cord and/or
moving the instrument.
6. Ensure that the power cord is firmly plugged into the power inlet, the wall outlet and AC adapter.
7. When the instrument is operating for a long time, its temperature can become too high. Please
be careful that the instrument’s temperature does not become too high during long and
continuous operation times. When operating, leave enough space around the instrument so there
is enough room for air circulation and cooling.
8. Do not disassemble the instrument in any event. If the instrument is malfunctioning or broken,
please contact your local distributor or Logos Biosystems. Disassembling the instrument
invalidates its warranty.
9. Use only authorized components (adaptor, power cord, and USB drive).
10. If the instrument emits smoke, disconnect the power cord immediately from the wall outlet and
contact a service person.
11. Do not reuse LUNA™ Cell Counting Slides. Used counting slides must be disposed as biohazard
waste according to the rules and regulations of your local government.

5
www.logosbio.com
<Symbols used in this User Manual>
TheWEEE(Waste Electrical and Electronic Equipment) symbol indicates that users
of this instrument have the responsibility of returning and disposing of WEEE in an
ecologically friendly manner. Follow waste ordinances of your region for proper
disposal provisions.
The CE mark indicates that this instrument conforms to all applicable European
Community provisions for which this marking is required. Users must be aware of
and follow the conditions described in this manual for operating the instrument. The
protection provided by the instrument may be impaired if the instrument is used in
a manner not specified by this manual.
Protective earth (Ground)

6
www.logosbio.com
General Guidelines for Using the LUNA™ Automated Cell Counter
In order to achieve the best results with the LUNA™ Automated Cell Counter, users must follow the
instructions below carefully.
1. The instrument must be operated in compliance with the operating environment described on
page 7. In particular, the temperature and humidity conditions are important.
2. Samples must be handled in an appropriate way, depending on user’s requirements.
3. Hold the LUNA™ Cell Counting Slide by the edges to avoid touching its optical surface. Make
sure that no damage or contamination occurs on the optical surfaces of the slide.
4. After mixing the cell sample with Trypan Blue Stain, perform cell counting as soon as possible,
within 3 minutes for accurate cell viability measurement. If needed, count your sample at least 2
times (duplicate readings) and take an average.
5. Since the LUNA™ Automated Cell Counter is calibrated before shipping, users generally do not
need to re-calibrate before use. However, if re-calibration is needed, please refer to Section 5.3
Calibrating the Counter.

7
www.logosbio.com
Environment Conditions
Operating Power
100 - 240 VAC, 1.5A
Frequency
50/60 Hz
Electrical Input
12 VDC, 3.5A
Installation Site
Indoor use only
Operating Temperature
10 - 35℃
Maximum Relative Humidity
20 - 80%
Altitude
≤2,000 m
Pollution Degree
2

8
www.logosbio.com
Chapter 1 –Introduction
1.1 Product Overview
The LUNA™ Automated Cell Counter is a small, fast, and affordable image-based cell counting
device that automatically counts various kinds of cells for research use.
The LUNA™ Automated Cell Counter helps measure the number, as well as viability of cells (live,
dead, total cells) with sophisticated optical components and advanced image analysis algorithms.
Due to several innovations introduced by Logos Biosystems, LUNA™ provides a state of the art cell
counting device and eliminates the tedium and subjectivity of manual cell counting.
The LUNA™ Automated Cell Counter can be used in a very simple procedure. First, mix 10 µl of the
cell sample with 10 µl of Trypan Blue Stain or Erythrosin B Stain. Second, load the cell suspension
into the LUNA™ Cell Counting Slide. Third, insert the slide into the slide port of the instrument and
adjust the focus knob to get an appropriate cell image. Last, press the “Count” button and the results
of cell count and viability will be displayed on the screen. The counting image can be downloaded
onto a USB drive in TIF format for future analysis.
The LUNA™ Automated Cell Counter provides key data as below:
Number of live and dead cells / mL
Number of total cells / mL
Viability percentage (% live cells to total cells)
Cell images (showing live cells as green circles and dead cells as red circles)
Histogram of cell size distribution
Both reusable and disposable slides can be used with the LUNA™. The LUNA™ Reusable Slide is
compatible with the LUNA™ family of automated cell counters for both brightfield and fluorescence
cell counting. Designed for cost-efficient an accurate cell counting, the LUNA™ Reusable Slide
combines the economy of manual cell counting with the speed, accuracy, and convenience of
automated cell counting. The disposable LUNA™ Cell Counting Slides maintain the highest standard
of cell counting accuracy and offer the ultimate counting experience with no mess or cleanup. Each
counting slide has 2 chambers, labeled as Aand B, so that one slide can be used for the same sample
reading in duplicate or for 2 different samples if preferred.

9
www.logosbio.com
Key features of the LUNA™ Automated Cell Counter are as follows:
Key features
Description
Small footprint
Due to its minimal size (22 x 21 x 8.5 cm) & weight (1.2 kg), it
can be used on a clean bench or stored in a biosafety cabinet
for convenience.
Accuracy & precision
Due to its sophisticated optical components and counting
algorithm, LUNA™ Automated Cell Counter provides optimized
and reproducible counting results every time.
Easy-to-operate user interface
The intuitive user interface based on a touch screen enables
simple and easy operation.
Shortest time-to-results
Results for most cell lines are available within 7 seconds of
pressing the ‘Count’ button.
Innovative counting slide
LUNA™ Automated Cell Counter adopts an innovative counting
slide made with “T-bond” technology without using hazardous
organic solvents.
Cell concentration & viability range
Measurements can be made for cells at concentrations within
the 1 x 104–1 x 107cells/mL range and for cells within the 3 -
60 µm diameter range.
Easy-to-use dilution calculator
Dilution concentration can easily be calculated using the built-
in calculator.
Setup & maintenance
Just plug in and it is ready for use, with virtually no maintenance
costs.
Counting image transfer
The counting image can be downloaded onto a USB drive in
TIFF (Tag Image File Format) for later use or review.
Individual protocol saving
Each researcher can set and adjust counting parameters such
as cell size and save the counting protocol for later use.
Automatic reporting
LUNA™ provides a counting report based on the count results
in PDF (Portable Document Format) immediately after
performing a cell count.
Cell information
Cell size, position, and viability information is provided for all
counted cells.

10
www.logosbio.com
1.2 Product Contents
The LUNA™ Automated Cell Counter in the product package consists of the following components.
Component
Quantity
LUNA™ Automated Cell Counter
1
Power cord (with AC adapter)
1
LUNA™ Cell Counting Slides, 50 Slides (100 Counts)
1 box
Trypan Blue Stain, 0.4 %
2 x 1 mL
USB Drive, 16 GB
1
When you receive the product package, please immediately check it with the above component list
to ensure that all parts are included and no damage has occurred during shipping. The warranty does
not cover damage that may occur during shipping and handling. Any damage claims must be filed
with the carrier.
Note: LUNA™ Automated Cell Counter is only for research use, not for human or animal therapeutic
or diagnostic use.

11
www.logosbio.com
1.3 Product Specifications
1.3.1. LUNA™ Automated Cell Counter specifications
Instrument Type
Benchtop cell counter
Dimensions (WxDxH)
22 x 21 x 8.5 cm
Weight
1.2 kg
Cell Concentration Range
1 x 104 –1 x 107cells/mL (Optimally 5 x 104 –1 x 107cells/mL)
Cell Diameter Range
5 - 60 µm
Cell Circularity Range
0 - 100%
Cell Viability Range
0 - 100%
Image Resolution
5 mega pixel (5MP)
Image Type
TIF (Optimized for LUNA™ only)
Software
LUNA™ software (www.logosbio.com)
Reporting
PDF format report
Processing Time
Approx. 7 seconds at 1 x 106cell/mL concentration
(Depending on cell type and concentration)
Note: For optimal cell viability measurement, the cell diameter of 9 µm or bigger is recommended,
depending on sample condition.
1.3.2 LUNA™ Cell Counting Slide specifications
Material
Polystyrene
Dimensions (WxDxH)
25 x 75 x 2.4 mm
Chamber Depth
100 µm
Chamber Volume
10 µl
Note: One USB Drive of 16 Gigabytes and 0.4% Trypan Blue Stain (2 x 1 mL) are included in the
starter package.

12
www.logosbio.com
1.4 Product Description
1.4.1 Front view of the LUNA™ Automated Cell Counter
The front view of the LUNA™ Automated Cell Counter shows a wide touch screen. This interface
contains buttons for all functions and displays results.
1.4.2 Rear view of the LUNA™ Automated Cell Counter
The rear view of the LUNA™ Automated Cell Counter shows a power button to turn on or off the
instrument and a power inlet. Connect the instrument to an electrical outlet with the power cord and
plug provided in the product package. Be sure to check the electrical outlet configuration in your
country.
Power inlet
Power button

13
www.logosbio.com
1.4.3 Right side view of the LUNA™ Automated Cell Counter
The right side view of the LUNA™ Automated Cell Counter shows a focus knob and a slide port. Use
the focus knob to get optimized cell images by adjusting contrast between live cells with bright centers
and dead cells with dark centers. The slide port is used to insert a compatible slide loaded with the
sample into the instrument.
1.4.4 Left side view of the LUNA™ Automated Cell Counter
On the left side of the cell counter, there is a USB drive port into which a USB drive is inserted for
transferring and saving data. One USB drive is supplied in the product package. Any standard USB
drive can be used.
Focus knob
Counting slide port
USB drive port

14
www.logosbio.com
1.4.5 LUNA™ Cell Counting Slide
The plastic disposable Cell Counting Slide consists of 2 chambers, labeled as Aand B, which can be
used for the same sample as duplicates or for 2 different samples. The depth of the counting chamber
is 100 µm. The counted cell volume is about 0.5 µl, almost the same as counting five (1 x 1 mm)
squares in a standard hemocytometer.
Sample loading chambers (marked as A and B)
Sample loading port

15
www.logosbio.com
Chapter 2 –Setting up
2.1 Installation
2.1.1 Upon receiving the product package, unpack it carefully and ensure that every component is
included and no damage has occurred.
2.1.2 Place the LUNA™ Automated Cell Counter on a flat, stable surface.
2.1.3 Insert one end of the power cord into the instrument and plug the other end of the power cord
into an electrical outlet after checking the outlet configuration in your local area.
2.1.4 Turn on the instrument using the power button located on the back of the cell counter.
2.1.5 The Start-up screen will show up on the touch screen as shown below in 2.2.

16
www.logosbio.com
2.2 Start-Up Screen
When the cell counter is turned on, the Start-Up screen is displayed. The Start-Up screen will show
7 compartments; menu, display, protocol & date, function, zoom, staining status, and setting.
<Start-Up Screen of the LUNA™ Automated Cell Counter>
Note: While operating the instrument, the buttons that are not in use are inactivated.
Note: The inner red box in the display screen above is the current viewing area. And, the outer red
box is the whole field of view of the LUNA. The size of the navigator (inner red box) is automatically
adjusted, depending on the zoom-in level selected by users as shown below.

17
www.logosbio.com
2.3 Settings
Generally, users do not need to change the “Settings” of the instrument since they are preset at the
time of manufacture. If users need to reset the date, time, or other options, users can adjust or change
the options/parameters from the “Settings” menu described below.
2.3.1 Press the “Settings” button ( ) located on the upper right corner of the counting screen. Then,
the following “Settings” screen appears. From here, users can change or adjust various preferences
or options.
<Settings>
2.3.3 The “Settings” menu allows you to perform the followings (which will be explained in detail
below):
Background calibration of the instrument (described in Section 5.3 Calibrating the Counter)
Firmware update for the installation of new versions of the firmware (described in Section
5.4 Updating the Firmware)
Date and time set up
Other options such as “Save”, “Counting”, and “Result/Printer/Calculator”

18
www.logosbio.com
2.4 Setting the Date and Time
2.4.1 On the “Settings” screen, select the date and time buttons.
2.4.2 After selecting the field, erase the number using the “Backspace” button ( ), set the date or
time using the number buttons, and press the “Apply” button ( ) to save the changes. .
2.5 Save Options
2.5.1 The “Save Options” screen will be seen as below, after selecting the “Save Options” button. As
a default, only the “Analyzed Image” button is activated (On), while the “Raw Image” and “Report”
button are inactivated (Off).
<Save Options>

19
www.logosbio.com
2.5.2 In the “Default Image Save” menu, users can choose their options to save the raw image, PDF
report, or both. Each button can be switched on and off.
Saved Items
Description
Analyzed image
The image contains the counting results and the tagging of live and dead
cells.
Raw image
The image contains only the captured image for counting. In the bright
field cell counting mode, the bright field image will be displayed.
Report
The PDF report containing counting results and histograms
IMPORTANT
Remember that the “Raw Image” MUST be turned on to save the “Raw Image”which
should be provided to your distributor or Logos Biosystems to get the best technical
support.
2.5.3 The “Auto-header” which is red-lined in the above “Save Options” image is the newly created
folder name or file name that all the images start with during the saving process. After saving, users
can see a folder or a file name, containing the saved item, which begins with “Auto-header” in the
root directory of the USB drive. In case users want to have another default name, the “Auto-header”
can be customized by editing the default value of “Logos Biosystems-”.

20
www.logosbio.com
2.6 Counting Options
2.6.1 In the “Counting Options”, users can adjust Staining Option and Auto Exposure function.
<Staining Options>
2.6.2 In the “Staining options” of “Counting Options”, users can choose one of the three staining
options as below.
Staining Options
Description
With Trypan Blue
(1:1)
This option can be used for normal bright field counting, when cell samples
are mixed with Trypan Blue dyes in a 1:1 ratio. This option can generate
cell viability data. For samples with Trypan Blue Stain, the dilution factor
must be set to value “2”. Please check.
Without Trypan Blue
When samples do not contain Trypan Blue dye, turn on this button and
follow the directions in the message boxes. These instructions set the
“Dilution Factor”. For samples with Trypan Blue dye, the dilution factor
must be set to value “2”. For samples without Trypan Blue dye, the dilution
factor must be set to value “1”.
Note: The cell counting parameters of the LUNA™ are optimized with the
use of Trypan Blue. Low contrast from no dye staining may sometimes
cause abnormal results.