NanoEnTek EVE User manual

User
Manual
EVE
™
Automatic cell counter

2
All the materials in this user guide are protected by Korean and
international copyright laws. They cannot be reproduced, translated,
published or distributed without the permission of the copyright owner.
EVE™ User manual
Website : www.nanoentek.com
E-mail : [email protected]
Manufactured by
NanoEnTek Inc. (HQ)
12F, 5, Digital-ro 26-gil, Guro-gu, Seoul, 152-740, Korea
Tel : [phone]
Fax: [phone]
The information in this manual is described as accurately as possible.
Firmware and software changes and updates may change without prior
consent or notification.
Copyright © 2013 by NanoEnTek Inc.
All rights reserved. Published in Korea.
Documentation : NESMU-EVE-001E
Revision history : V.0.0 Date : MAR 2013
: V.0.5 Date : MAY 2013
: V.0.8 Date : OCT 2013
: V.1.5 Date : MAY 2014

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 3
Table of contents
Product contents
Product overview
Description
Installation
Operation
Settings
Cell mode parameters
Date and time setup
Recommend actions
Cell counting
Transferring data to a computer
Using the beads mode for counting beads
Calibrating
PC software
Installation
Open the data
Cell gating
Counting
Spread sheet
Prepare report
Cleaning and maintenance
Troubleshooting
Error codes
Warranty
Safety precautions
Product specifications
Ordering information
Technical support
4
5-6
7-8
9
10-25
10
10-12
13
14
15-20
21
22-23
24-25
26-33
26-27
28-29
30
31
32
33
34
35-36
37
38
39-41
42
43
44

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Product contents
EVE™ is shipped with the following components.
Please check that all items listed below were shipped, receiving the instrument.
If any items are missing or damaged, contact your local distributor or e-mail
[email protected].
Automatic cell counter
1 EA
USB drive
4 GB
1 EA
User manual
1 EA
Power cord with 4 adaptor
cords ( 4 pcs/set, for U.S./Canada/
Taiwan/Japan, Europe or UK)
1 SET
Cell counting slides
(with 1.5 mL of Trypan blue (0.4%))
1 BOX (50 slides/box)
Quick manual
1 EA
Cat. No. EVE-MC
Cat. No.
EVS-050

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 5
Product overview
The EVE™ Automatic cell counter uses state-of-the-art optics and image
analysis to automatic cell counting. The EVE™ is a benchtop counter designed to
measure cell count and viability (live, dead, and total cells) accurately and precisely,
using the standard trypan blue technique.
Using the same amount of sample that you currently use with the hemocytometer, the
EVE™ takes less than 20 seconds per sample for a typical cell count and is compatible
with a wide variety of eukaryotic cells and provides information on cell size.
The EVE™ offers an intuitive user interface, and provides the option to save and print
cell count data using the EVE™ Software (download from www.nanoentek.com )and
USB drive supplied with the instrument or available separately.
The EVE™ is supplied with disposable EVE™ Cell counting slides that contain two
enclosed chambers to hold the sample to allow you to measure two different samples or
perform replicates of the same sample. The cell counting occurs in the central location
of the counting chamber and the volume counted is 0.4 µL, the same as counting four
(1 mm × 1 mm) squares in a standard hemocytometer.

6
Product overview
Features and benefits
—
User-friendly, benchtop design for simple, fast, automated cell count and
viability measurements within 20 seconds.
—
Provides data on cell size and is compatible with a wide variety of eukaryotic cells
without the need for any special changes between large or small sizes.
—
Measures cell concentrations ranging from 1 × 104to 1 × 107cells/mL and
cells with sizes ranging from 5 µm to 60 µm.
—
Provide the clumping cell counting function to get more accurate results
(using EVETM PC software).
—
Uses disposable cell counting slides that eliminate washing steps and
cross contamination between samples.
—
Saves and print cell count data including images using the EVE™ software and
EVE™ USB drive.
—
Presents comprehensive data with graphical reports and as a .CSV
(comma separated value) file for sample comparisons.

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 7
Description
Front view The power button is used to turn the instrument
on and off. The red status light indicates that
the instrument is off; the green status light
indicates that the instrument is on.
Located in the front of the instrument contains
buttons for all the functions needed and
displays data from the cell count.
The slide port is used to insert the EVE™
Cell counting slide containing the sample with
trypan blue stain into the counter for analysis.
The USB port allows you to transfer and save
the cell count data and image to your computer
for record keeping and printing purposes. The
USB drive supplied with the instrument or any
other standard USB drive is inserted into the
USB port for data transfer.
Power button
LCD touch screen
Slide port
USB port
[Front view]
Power button
Slide port
USB port
LCD
touch screen

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Description
Rear and
side view
Connect the counter to an electrical outlet
using the supplied power cord and the
appropriate plug, based on the electrical outlet
configuration in your country.
The image adjustment (focus) knob is used
to adjust the image quality to obtain better
contrast between live (bright centers) and dead
(dark blue centers) cells. This is important
to obtain accurate cell counts and viability
measurements.
The focus lock knob is used to lock the image
adjustment (focus) knob once the image is
optimized. There is no need to use the focus
lock knob , but is available for convenience,
if you are measuring multiple samples of the
same cell type.
Power inlet
Image adjustment
(focus) knob
Focus lock knob
[Rear view] [Side view]
Power inlet Focus lock Focus knob

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 9
1. After unpacking the instrument, place the instrument on a
flat, level, dry surface.
3. Plug the power cord into the electrical outlet. Be sure to use
only the power cord supplied with your instrument. Powering the
instrument with an unapproved power cord may damage the
instrument.
5. When the instrument is turned on, the Start Up screen is
displayed. Here you can proceed immediately to cell counting,
set up the instrument for cell or bead count, or adjust the screen
brightness.
Installation
Installing
EVE™
Automatic
cell counter
2. Plug one end of the supplied
power cord into the EVE™. To the
other end attach the appropriate plug
adaptor, based on the electrical outlet
configuration in your country.
4. When you are ready to use, start the EVE™ by pressing the
Power button.

10
Operation
Settings
Cell mode
parameters
1. Press Settings from the Start-Up screen to display Settings.
The settings menu allows you to set up the following:
· Count mode to select Counting type and Auto focusing options. The Counting
type is to operate an instrument for cell counting or beads counting. The Auto
focusing is to operate an instrument for AF on mode or AF off mode.
· Count mode to operate the instrument for cell counting (choose Cells
or bead counting (choose Beads)
· Parameters (see below and next page for details)
· Calibration to calibrate the instrument (page 24)
· Update to install new firmware versions as they become available
· Date and Time to set up date and time (page 13)
· Use the scroll buttons to adjust the screen brightness.
1. Press Parameters from the settings screen to display Cell mode
parameters screen.
The Parameters function allows you to change the image analysis algorithm
for specific or mixed cell types, and the specific parameters must be
determined empirically.

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 11
Operation
Cell mode
parameters
2. The cell mode parameters are described below:
∙ Sensitivity (refers to the contrast of the objects from the background).
Adjusting the sensitivity higher makes instrument more sensitive to
objects; useful for cells that do not stain well with trypan blue while
adjusting the sensitivity lower makes the instrument less sensitive and is
useful if there is a lot of background.
∙ Minimum cell size is used to determine the low range of cell size to
include in the measurement. The algorithm first identifies all objects, and
calculates the average size (e.g., 15 µm). From the percent of average
size setting, the algorithm calculatesthe smallest object size to include in
the final measurement (e.g.,70% of 15 is10.5 µm; 15–10.5=4.5 µm;4.5
µm would be the smallest particle included in the count). Adjusting the
number up, increases inclusiveness thereby decreasing the lower cell
size range (e.g., 50% of 15 is 7.5 µm; 15–7.5 = 7.5 µm).
∙ Maximum cell size is used to determine the high range of cell size to
include in the measurement. The algorithm first identifies all objects, then
calculates the average size (e.g., 15 µm). From the percent of average
size setting, the algorithm calculates the largest object size to include in
the final measurement (e.g., 200%; 200% of 15 µm = 30 µm; 30 µm is
the largest cell size included in the measurement.
∙ Circularity is used to determine the objects to include in the
measurement based on roundness. Increasing the value from 80%
requires objects to be more round for inclusion in the measurement.
Decreasing the value from 80% allows objects to be less round. Adjusting
this may be useful if the cell type is not particularly circular or perhaps
oddly shaped.
After modifying any parameters,
press Apply button to make the changes.
To restore default parameters, press Default button.
3. Press Save protocol button
to create a new protocol.

12
Operation
Cell mode
parameters
4. Type protocol name and user name in the appropriate
fields, and press Save button again.
5. Once a protocol is saved, it is available for use at any time.
Press Load protocol button.
6. The protocol appears in the protocol menu. Use the up and
down arrows to find your saved protocol.
To use the protocol, select one of protocol and press Apply
button.
»press Delete button to delete protocol.
press Edit button to edit protocol.

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 13
Operation
Date and
time setup
1. Turn on the EVE™ by pressing the Power button. The Start
up screen is displayed after a few seconds.
2. Press Settings and then press Date and Time.
3. The Date/Time Properties screen is displayed. To select the
month and day, scroll to the appropriate month using the arrow
keys and then press the day on the calendar.
» Use a pointed object, like a stylus or pipette tip, to push the small buttons on the
calendar.
4. To select the time, scroll to the appropriate time and select
Automatically adjust clock for daylight saving, if necessary.
Press Apply or OK button to make the date/time changes. The
updated date/time is displayed on the top of the window.
Once the date/time is set, there is no need to set it each time the
instrument is turned on.
5. Press Close button to exit the screen.

14
Operation
Recommend
actions
To obtain the best results, follow these recommendations:
∙Wear Protective gloves during sample handling.
∙Do not touch the optical surfaces of the EVE™ Cell counting slides.
Hold the cell counting slides by the edges.
∙Use the EVE™ at room temperature only (5 - 40 °C).
∙For accurate viability count results, ensure the counting area is covered
with cell suspension and count cells within 3 minutes of mixing the cells
with trypan blue solution as trypan blue is toxic to cells. For best data
with biological samples, we recommend counting at least two samples
and taking an average.
∙The EVE™ is supplied pre-calibrated. To recalibrate your instrument,
see page 24.
∙The EVE™ memory holds one set of data. Save your data to the USB
drive after each reading.You may transfer the data to your PC, using the
USB drive immediately as described in Transferring data to a PC (page
21).
∙After using EVE™, appropriately dispose slides as biohazardous waste.
Do not reuse the cell counting slides.

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 15
Operation
Cell counting 1. Push the Power button to start the instrument. The Start-up
screen is displayed.
2. The EVE™ is preset to Cells mode.
3. Mix well the 10 µL of your sample and the 0.4% trypan blue
stain using a pipette.
4. Load 10 µL of the sample mixture on EVE™ Cell counting
slide (side A) using a pipette.
The two chambers of the slide are labeled “A” and “B” for easy
tracking of your samples.

16
Operation
Cell counting 5. Insert the EVE™ Cell counting slide, sample side (side A) first into
the slide port on the instrument, you will hear a soft click, if the slide is
pushed in correctly. Each chamber is counted separately.
☞ Auto focusing (AF on) mode is set as a default.
If necessary to personalize focus, turn on the manual focusing (AF off) mode in the
Setting menu (Refer to page 10).
[In case of Auto focusing (AF on) mode]
6-1. Press the Count cells.
6-2. The counting process will be started in the Auto Focusing (AF on)
mode.
☞For information on how to check counting results, refer to the page 19.

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 17
Operation
Cell counting [In case of Manual focusing (AF off) mode]
7-1. Adjust the image by pressing the Zoom button. Navigate by
pressing the location you like to see on the grid
7-2. While viewing cells in the zoom mode, use the Focus knob to
adjust the image.
☞For information on how to check counting results, refer to the
page 19.
» After you have counted the first
sample, you may not have to use
the Focus knob again. If measuring
multiple samples of the same
approximate size, you may lock the
Focus knob. You are able to unlock
the knob later, to adjust the image.

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Operation
Cell counting Optimize the image for analysis such that:
∙Live cells have bright centers and dark edges.
∙Dead cells have a uniform blue color throughout the cell with
no bright centers.
<Correct Image>
<Incorrect Image 1>
Dead cells have bright, blue centers
are counted as the live cells.
<Incorrect Image 2>
Live cells have dark centers and are
counted as the dead cells.
67.733 mm

EVETM, Automatic Cell Counter ©2013 NanoEnTek Inc. 19
Operation
Cell counting 9. When you are satisfied with the image, press Count Cells.
10. After 20 seconds to count each sample, and the cell count for
live, dead, and total cells as well as percentage viability is displayed
on the screen. Record the cell count, or insert a USB drive and press
Save button (see page 21).
11. To see more details on the data as well as graphical
representation of the data, press the More data button.
» Press the Close button to return to the main screen.
67.733 mm

20
Operation
Cell counting 12. The Calculator button allows you to quickly calculate
adjustments to the cell suspension to obtain a desired
concentration.
13. To count the cells in the other side of the slide (side B),
remove the EVE™ slide after side A is counted by pushing in the
slide slightly and then pulling the slide out. Turn the slide around
and reinsert into the slide inlet and repeat the procedure.
14. The EVE™ USB drive holds one set of data. To save your
data for future analysis or archiving, you must record the data or
save after each reading.
See page 21 for Transferring data to a PC.
15. After recording or saving the data, remove and discard the
slide appropriately as bio-hazardous waste.
16. At this point, the EVE™ is ready for another sample. If you
are not using the instrument, press the Power button to turn off
the instrument.
» If the touch screen is not responding, you can turn off the instrument by pressing
and holding the Power button for 4 seconds.