logos biosystems Luna II User manual

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Catalog No.: L40001 / L40002
Document No.: L44001
Revision Date: December 02, 2014
SW Version: 1.4.4

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IMPORTANT
The LUNA-II™ Automated Cell Counter is a Laboratory Electrical Instrument for
Scientific Research Use Only, BUT NOT A MEDICAL or THERAPEUTIC or IN
VITRO DIAGNOTICS DEVICE.

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Table of Contents
Safety Information ----------------------------------------------------------------------------------------
5
General Guidelines --------------------------------------------------------------------------------------
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Environment Conditions --------------------------------------------------------------------------------
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Chapter 1 –Introduction ----------------------------------------------------------------------
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1.1 Product Overview --------------------------------------------------------------------------------
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1.2 Key Features --------------------------------------------------------------------------------------
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1.3 Product Contents ---------------------------------------------------------------------------------
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1.4 Product Specifications --------------------------------------------------------------------------
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1.5 Product Description ------------------------------------------------------------------------------
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Chapter 2 –Setting up --------------------------------------------------------------------------
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2.1 Installation ------------------------------------------------------------------------------------------
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2.2 Start-Up Screen -----------------------------------------------------------------------------------
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2.3 Settings ---------------------------------------------------------------------------------------------
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2.4 Calibrating the Touchscreen (Calibrate Touchscreen) ----------------------------------
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2.5 Changing Options for Cell Counting (Counting Options) -------------------------------
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2.6 Setting the Date and Time ---------------------------------------------------------------------
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2.7 Calibrating LUNA-II™ (Calibrate) ------------------------------------------------------------
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2.8 Updating the Software (Update Software) -------------------------------------------------
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Chapter 3 –Protocol Setting ----------------------------------------------------------------
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3.1 Parameters in the Protocol ---------------------------------------------------------------------
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3.2 Setting the Protocols ----------------------------------------------------------------------------
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3.3 Selection of Protocol -----------------------------------------------------------------------------
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Chapter 4 –Counting Cells ------------------------------------------------------------------
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4.1 Preparing the Instrument -----------------------------------------------------------------------
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4.2 Sample Preparation ------------------------------------------------------------------------------
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4.3 Loading Samples into the Luna™Cell Counting Slide ---------------------------------
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4.4 Counting Cells -------------------------------------------------------------------------------------
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4.5 After Counting: Image View --------------------------------------------------------------------
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4.6 After Counting: Histogram & Gating ---------------------------------------------------------
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4.7 After Counting: Saving and Printing the Results ------------------------------------------
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4.8 After Counting: Calculation for Subsequent Experiments ------------------------------
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Chapter 5 –Focusing Option ---------------------------------------------------------------
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5.1 Autofocusing ---------------------------------------------------------------------------------------
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5.2 Manual Focusing ---------------------------------------------------------------------------------
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Chapter 6 –Review the Previous Results ------------------------------------------
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Chapter 7 –Maintenance and Troubleshooting ---------------------------------
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7.1 Turn On/Off LUNA-II™ --------------------------------------------------------------------------
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7.2 Cleaning --------------------------------------------------------------------------------------------
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7.3 Calibrating the Touchscreen -------------------------------------------------------------------
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7.4 Calibrating LUNA-II™ ---------------------------------------------------------------------------
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7.5 Updating the Software --------------------------------------------------------------------------
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7.6 Troubleshooting -----------------------------------------------------------------------------------
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Chapter 8 - Ordering Information --------------------------------------------------------
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Chapter 9 - Purchaser Notification ------------------------------------------------------
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9.1 Limited Use Label License ---------------------------------------------------------------------
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9.2 Instrument Warranty -----------------------------------------------------------------------------
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Contact Information ------------------------------------------------------------------------------
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Safety Information
For best results with the LUNA-II™ Automated Cell Counter, follow the instructions below in addition
to the general precautions for using electrical instruments.
1. 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 9.
2. Trypan blue stain is known as a hazardous material. While handling the solution, always wear
proper personal protective equipment (PPE) 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 surface 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 out of order or dropped or
broken, please contact a service person. 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. Used counting slides must be disposed as biohazard wastes.

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<Symbols used in this User Manual>
The WEEE (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)

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General Guidelines for Using the LUNA-II™ Automated Cell
Counter
In order to achieve the best results with the LUNA-II™ Automated Cell Counter, follow the
instructions below carefully.
1. The instrument must be operated in compliance with the operating environment described on
page 9. 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-II™ Automated Cell Counter is calibrated before shipping, you do not need to
re-calibrate before use. However, if re-calibration is needed, please refer to Section 5.3
Calibrating the Counter.
6. Do not touch trypan blue solution with bare hands as it is a hazardous chemical. After using the
counting slides, dispose of it as hazardous wastes. Do not reuse the slides.

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Environment Conditions
Operating Power
100 ‒240 VAC, 1.2 A
Frequency
50/60 Hz
Electrical Input
12 VDC, 3.3 A
Installation Site
Indoor use only
Operating Temperature
10 ‒35℃
Maximum Relative Humidity
20 ‒80%
Altitude
≤2,000 m
Pollution Degree
2

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Chapter 1 –Introduction
1.1 Product Overview
The LUNA-II™ Automated Cell Counter is a small, fast, and affordable image-based cell counting
device that automatically counts various kinds of cells for research purpose.
The LUNA-II™ 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-II™ provides a state of the art
cell counting device and eliminates the tedium and subjectivity of manual cell counting.
The LUNA-II™ Automated Cell Counter can be used in a very simple procedure. First, mix 10 l of
the cell sample with 10 l of trypan blue stain. Second, load the cell suspension into the Luna™ Cell
Counting Slide. Third, insert the slide into the slide port of the instrument and chose [Autofocused
Counting] or adjust the focus by clicking arrows to get an appropriate cell image. Last, press the
[Count] button and then 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 review and/or record keeping.
The LUNA-II™ 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)
Histograms of cell size distributions
The Luna™ Cell Counting Slide is disposable and specifically designed for the LUNA-II™
Automated Cell Counter. Each counting slide has 2 chambers, labeled as A and B, respectively, so
that one slide can be used for the same sample reading in duplicate or for 2 different samples if
preferred.

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1.2 Key Features
Key features
Description
Small footprint
Compact size with light weight saves space and is suitable for
either laboratory table or biosafety cabinet.
Accuracy & precision
With sophisticated optical components and counting algorithm,
LUNA-II™ provides optimized and reproducible results every
time.
Autofocusing
Advanced liquid lens technology provides fast autofocusing
without mechanical moving. Autofocusing enables reliable cell
counting and excludes human error.
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 15 (without
autofocusing) or 22 (with autofocusing) seconds after pressing
the [Count].
Built-in printer (Optional)
An integrated thermal printer provides concise Cell Count
Report facilitating record keeping.
Innovative counting slide
LUNA-II™ 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
ranging from 5 x 104to 1 x 107cells/ml and for cells within the
3 ‒60 m diameter range.
Easy-to-use calculator
No external calculator is needed to calculate subsequent
dilution.
Setup & maintenance
Just plug in and it is ready for use, with virtually no
maintenance costs.
Counting image acquisition
The captured image of cells can be downloaded onto a USB
drive in TIFF (Tag Image File Format) for review or record
keeping.
Individual protocol
Different protocols (up to 300 protocols) can be saved with
personalized parameters.
Documentation
LUNA-II™ provides a PDF (Portable Document Format) report
which includes File name, Date, Cell count results, Cell
images, and histograms.

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1.3 Product Contents
The product package of the LUNA-II™ Automated Cell Counter contains the following components.
Component
Quantity
LUNA-II™ Automated Cell Counter
(with or without printer)
1
Power cord (including an adapter)
1
Luna™ Cell Counting Slides
1 box (50 slides for 100 counts)
Trypan Blue Stain (0.4 %)
2 x 1 ml
Luna™ USB drive
1
After receiving the product package, please immediately unpack it and check the components listed
above to ensure that all parts are included and no damage has been 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: The LUNA-II™ Automated Cell Counter is only for research purpose, not for human or animal
therapeutic or diagnostic use.

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1.4 Product Specifications
1.4.1. LUNA-II™ Automated Cell Counter specifications
Instrument Type
Benchtop cell counter
Dimensions (WxDxH)
16 x18 x28 cm (6.3 x7.0 x11.0 inch)
Weight
1.6 kg (3.5 lb) without the external power adaptor
Cell Concentration Range
5 x 104 ‒1 x 107cells/ml
Cell Diameter Range
3 ‒60 m (optimal 8-30 m)
Cell Viability Range
0 ‒100%
Image Resolution
5 mega pixels (5 MP)
Image Type
TIF format (Optimized for LUNA-II™only)
Software
LUNA-II™ software (www.logosbio.com)
Documentation
PDF report
Processing Time
Nominal time for cell counting is less than 10 (without
autofocusing) or 15 (with autofocusing) seconds at ~1 x 106
cell/ml concentration.
(Processing time may vary by cell type and concentration)
1.4.2 Luna™ Cell Counting Slide specifications
Material
Polystyrene
Dimensions (WxDxH)
25 x 75x 2.4 mm
Chamber Depth
100 m
Chamber Volume
10 l
Note: One Luna™ USB Drive (4 Gigabytes) and 0.4% Trypan Blue Stain solution (2 x 1 ml) are
included in the starter package.

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1.5 Product Description
1.5.1 Front view of the LUNA-II™ Automated Cell Counter
The front view of the LUNA-II™ Automated Cell Counter shows various parts as shown below.
Wide touchscreen located in the upper front of the instruments contains buttons for all functions
needed to operate the instrument and displays data acquired.
Power button is used to turn on the instrument.
Counting slide port is used to insert the Luna™ Cell Counting Slide containing sample to
analyze.
The front USB port provides easy-to-access data retrieving to transfer.
1.5.2 Rear view of the LUNA-II™ Automated Cell Counter
The rear view of the LUNA-II™ Automated Cell Counter shows two additional USB ports and a
power inlet to connect the instrument to an electrical outlet with the power cord and plug which are
provided in the product package. Be sure to check the electrical outlet configuration in your country.

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1.5.3 Right side view of the LUNA-II™ Automated Cell Counter
The right side view of the LUNA-II™ Automated Cell Counter shows a built-in printer (L40001 model
only) which allows rapid printing the cell counting results.
1.5.5 Luna™ Cell Counting Slide
The Luna™Cell Counting Slide is a plastic disposable cell counting slide consists of 2 chambers,
labeled as A or B, that can be used for the same sample as duplicate or for 2 different samples. The
depth of the counting chamber is 100 m. The volume of cells counted is about 0.5 l, almost the
same as five (1 mm x 1 mm) squares in a standard hemocytometer.

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Chapter 2 –Setting up
2.1 Installation
Upon receiving the product package, unpack it carefully and ensure that every component is
included and no damage has been occurred.
Place the LUNA-II™ Automated Cell Counter on a flat and stable surface.
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.
Turn on the instrument using the power button located on the front of the instrument. Initializing
screen after company logo will be displayed a few seconds as below.
Company logo
Initializing screen of the LUNA-II™ Automated Cell Counter. In the initializing
screen, the current version of LUNA-II™ OS can be identified in the right lower
corner. Initializing screen will be followed by LOADING sign as below.

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LOADING sign of the LUNA-II™ Automated Cell Counter.
The Start-Up screen will be displayed on the touchscreen as shown below in 2.2.
See Section 7.1 for Turn on and Turn off the instrument.

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2.2 Start-Up Screen
The Start-Up screen will show 5 compartments; Power icon, count, review, protocol, and settings.
The Power icon in the Start-Up screen can be used to turn off the instrument. See Section 7.1. for
further information to turn on/off the instrument.

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2.3 Settings
Generally, there is no 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 in the “Settings”menu described below.
Press [settings] located on the right of the Start-Up screen as below.
Then, the following Settings screen will be displayed.
In the Settings screen, users can see the followings:
Home image: by Pressing Home image, you can go to the Start-Up screen.
Current protocol and date in the upper right corner.
Date and values of last calibration.
Date and software version of latest software update.

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The Settings menu allows you to perform the followings:
Change options for cell counting by pressing [Counting Options].
Set the date and/or time by pressing [Date / Time].
Re-calibrate the instrument by pressing [Calibrate].
Update the software by pressing [Update Software].
Calibrate the touchscreen by pressing [Calibrate Touchscreen].

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2.4 Calibrating the Touchscreen (Calibrate Touchscreen)
This function can be used when the response of touchscreen is abnormal.
On the Settings screen, press [Calibrate Touchscreen] as below.
Now, users can see the following screen. If users want to calibrate the touchscreen, press [OK]. If
not, press [Cancel].