CORNING Cell Counter Manual de usuario

Corning®Cell Counter

Accurate Cell Counts in a Flash

Instruction Manual

Cat. No. 6749

CORNING CELL COUNTER Instruction Manual

Table of Contents

Safety Information ............................................................4

1.0 Introduction ..............................................................5

Technology to Make Your Work Easier ......................................5

How Easy the Corning® Cell Counter Works .................................5

Operating Range ..........................................................5

Instrument Overview ......................................................6

2.0 Getting Started ...........................................................7

3.0 Setting Up the Corning Cell Counter .......................................8

Setup Procedure...........................................................8

Registration of the Device .................................................9

Error Codes ............................................................. 10

Start a Project........................................................... 11

Start a Cell Count ........................................................12

Adapter Plate Placement .................................................12

Sample Loading ..........................................................13

Live/Dead Sample Loading................................................14

Counting Chamber Placement ............................................14

4.0 Performing a Cell Count ..................................................15

Focusing of the Cells......................................................15

Performing a Count ......................................................16

5.0 Connecting to the Cloud ..................................................17

Confirmation Email.......................................................17

Project Overview .........................................................18

Experiment Overview.....................................................19

Experiment Results.......................................................20

6.0 Cleaning and Troubleshooting ............................................21

Cleaning and Maintenance ...............................................21

Troubleshooting..........................................................21

USB device does not connect ...........................................21

No cells are visible .....................................................22

Too many cells ....................................................... 22

Cell clumps............................................................22

Bubbles ...............................................................23

Debris.................................................................23

7.0 Technical Specifications...................................................24

8.0 Limited Warranty.........................................................25

9.0 Equipment Disposal ......................................................26

Contact Information ................................................ Back cover

CORNING CELL COUNTER Instruction Manual

Safety Information

To avoid damage or malfunction of the Corning® Cell Counter:

 Be sure to set this device on a firm, stable, and horizontal surface.

 Shut down the device if it produces smoke, a strange odor or unusual noise. Continued

use may lead to fire. Unplug the USB cable immediately.

 Do not connect the USB cable in ways other than those mentioned in this manual.

 If water or other liquid enters the device, discontinue use immediately. Continued use

may lead to fire. Unplug the USB cable immediately.

 Do not attempt to repair or remove the outer case of the device.

 Never attempt to repair, disassemble, or modify this device yourself, as tampering

with this product may result in injury or fire.

 Do not drop the device or allow objects to fall onto it.

 Do not allow the device to be submerged in liquid.

 Do not place heavy objects on top of this device, and never stand or lean on this

device. Failure to follow this may cause the equipment to fall or collapse, which may

result in possible breakage and/or injury.

 Do not autoclave this device.

 Do not perform the sterilization cycle of an incubator with the device inside.

 Do not use acetone to clean the device.

1.0 Introduction

Technology to Make Your Work Easier

Thank you for purchasing the Corning® Cell Counter.

The Corning Cell Counter is a benchtop automated image-based cell counter, which

uses state of the art optics and image analysis software. Its innovative design combines

the best qualities of manual cell counting with those of automated image-based cell

counting. With this, the Corning Cell Counter can accurately and precisely measure cell

concentrations and viability – live, dead, and total cells – using the standard Trypan blue

technique.

How Easy the Corning Cell Counter Works

The Corning Cell Counter system can be installed within several minutes and requires

minimal training.

The Corning Cell Counter uses a standard counting chamber or customer supplied

hemocytometer to analyze cell samples. Images from the cell sample are obtained by

CytoSMART™ Cloud App analyses of these images in the cloud. Sophisticated image

analysis algorithms produce accurate cell counts within a few seconds.

You can directly obtain your cell concentration on the results page or look it up later on

the CytoSMART™ Connect.

Operating Range

The Corning Cell Counter can be used to determine cell concentrations of mammalian

cells. The system works within a cell size range between 5.0 to 70 µm. Expect accurate

results for concentration samples within a range of 5.0 x 105to 1.0 x 107cells/mL.

CORNING CELL COUNTER Instruction Manual

Instrument Overview

The hardware of the Corning® Cell Counter is shown below (Figure 1):

1. Light arc – Contains the light source for imaging

2. USB cable – For connecting the device via USB, which is also the power source for

the device

3. Optical window – Cover glass on which the counting chamber is placed

4. Adapter plate – Used to guide the counting chamber across the optical window

5. Focus knob – Allows for changing the focus of the image

Figure 1. Instrument overview of the Corning Cell Counter

1. Light Arc

2. USB Cable

5. Focus Knob

3. Optical Window

4. Adapter Plate

2.0 Getting Started

Product contents include the following items (Figure 2):

1. Manual and Quick Starter Guide

2. Corning® Cell Counter

3. License key

4. Adapter plate for the counting chamber

5. Counting chamber with two cover slips

6. Download link to the CytoSMART™ Cloud App software

7. USB drive for software

Figure 2. Package overview

Manual Corning Cell Counter Adapter plate for

counting chamber

License keyCounting chamber

with two cover slips

For additional information or any questions regarding your purchase, visit

[email protected] or contact our support staff at [email protected].

CORNING CELL COUNTER Instruction Manual

3.0 Setting Up the Corning®Cell Counter

To set up the Corning Cell Counter, the following items are needed:

 A Windows® 10 computer with internet access (preferably via a LAN-cable), and a USB

port (preferably USB 3.0). The computer or tablet must be on AC power, rather than on

battery power.

 The Corning Cell Counter

 The download link for the CytoSMART™ Cloud App software

 A license key to activate the device within the software

Setup Procedure

1. Open an internet browser and download the CytoSMART Cloud App using the

download link provided. If no link has been provided, please contact your sales

representative or contact our support staff via [email protected].

2. When the download is finished, double click the file to install the CytoSMART Cloud

App on the Windows computer.

3. Be sure to remove any wrapping before first time use of the Corning Cell Counter.

Also clean the device with a damp, lint-free wipe to clean the outer case with

70% ethanol or isopropanol. After the installation has completed, an icon for the

CytoSMART Cloud App will be visible on the desktop (Figure 3).

4. Start the CytoSMART Cloud App by double clicking the icon on the desktop. A

notification to connect the device will appear (Figure 4).

5. Next, connect the cleaned Corning Cell Counter to the Windows computer using

the USB cable. Connect via USB 3.0 port, if possible. This is required for optimal

performance.

6. When the Corning Cell Counter is connected, the green LED will activate in the light

arc. This indicates that the device is ready for use.

Figure 3. CytoSMART

Cloud App icon Figure 4. Connect the device notification

Registration of the Device

Before you can use the Corning® Cell Counter, you must register it. By clicking on

the “Register a new user” button (Figure 5), you will be led to the registration form

(Figure 6). Please fill in all the details and click “Sign up”.

Figure 5. Create an account

Figure 6. Sign up registration form

Figure 7. Register a device

The next step is to register the device. In the product box, you will find the license key

for this device. Enter the license key that is provided in the field “License” and click

“Register” (Figure 7).

Congratulations, you have now successfully registered and activated your Corning

Cell Counter!

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CORNING CELL COUNTER Instruction Manual

Error Code: Multiple Customer Environments

Is your personal account linked to more than one company? In that case, you will see

the comment box as shown in Figure 8 after signing in with your email address and

password. Select the desired environment that you would like to use from the drop-down

menu and click “Select”. This will lead you to the right environment to start your project.

Signing In

If your device is already activated and you already have an account, you can easily sign

in. Fill in your email address and password, and click “Log in” (Figure 9).

Figure 10. Warning: account not connected to environment

Figure 9. Sign in

Figure 8. Select an environment

Start a Project

With the Corning® Cell Counter you can perform a single count or multiple counts

within a project. The CytoSMART™ Cloud App software automatically stores the results

within a project, which you can view later in the CytoSMART Connect.

Figure 11. Create a project

Figure 12. Start a project

Error Code: Wrong Account or Company

If your personal account is not connected to the environment of the device, you will see

a warning (Figure 10). This means that the device is already activated and registered to a

company, but your personal account is not linked to this company.

After selecting “Create a new project” (Figure 11), you will be led to the “Start a project”

page (Figure 12). Next to the project owner details, which are already filled in, you can

also fill in the email address of collaborating users such as your colleagues. Name the

project, and give it a description. The final step is to assign a colored icon to the project.

This helps you to find your project easily, based on the color of the icon.

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CORNING CELL COUNTER Instruction Manual

Start a Cell Count

To start a cell count, you need to create an experiment. First, select

the project that your experiment belongs to using the drop-down

menu (Figure 11). If the cell count doesn’t belong to any of the

projects shown, click “Create a new project” to create a new project.

Next, give the cell count a name. We recommend always including

the date of the cell count in the file name to easily find the

experiment later on. After entering the details, click “Start cell

count” (Figure 13) and count your cells!

Adapter Plate Placement

You can use the adapter plate to guide the counting chamber during image acquisition

of the sample. We recommend to place the adapter plate on the Corning® Cell Counter,

but this is not required to perform a cell count. Using the adapter plate will produce the

most reliable results, as it allows for precise and repeatable handling of the counting

chamber.

To install the adapter plate:

 Make sure that both the Corning Cell Counter and the adapter plate are clear of any

debris. Note: Use a lint-free wipe to clean the device, if needed.

 Carefully slide the notch of the adapter plate around the base of the light arc (Figure 14).

 Push the adapter plate downwards, until it clicks into place on-top of the Corning

Cell Counter (Figure 15).

Figure 13. Start

cell count

Figure 14. Slide the

adapter plate into place.

Figure 15. Carefully press the

adapter plate downwards.

 Ensure the adapter plate is correctly placed. If it is placed incorrectly, this can lead to

false results (Figure 16).

 To remove the adapter plate, carefully pull it upwards near the base of the light arc.

Then carefully slide the plate from the Corning® Cell Counter.

Figure 16. Correct adapter plate placement

Sample Loading

A counting chamber is used for cell sample analysis. Most common hemocytometers are

compatible with the Corning Cell Counter. This is because these counting chambers have

a similar counting chamber depth of 0.1 mm.

The Corning Cell Counter does not require the counting grid – essential for manual

counting – to be included to perform a cell count. Therefore, you can also use disposable

counting slides, if they have a counting chamber depth of 0.1 mm.

Load the counting chamber with a cell sample according to your loading protocol. We

recommend loading 10 µL of the sample in the counting chamber with a 10 µL or 20 µL

pipet. The Corning Cell Counter operates within a concentration range of 5 x 105to

1 x 107cells/mL. If the cell solution is out of this concentration range, use Phosphate

Buffered Saline (PBS) or medium to resuspend the cells in an appropriate volume;

otherwise, you may obtain false results.

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CORNING CELL COUNTER Instruction Manual

Live/Dead Sample Loading

To perform a live/dead analysis, we recommend using the standard Trypan blue

technique. Mix a minimum of 20 µL sample with 20 µL trypan blue by using a 20 µL

pipet. Next load 10 µL of the sample mixture into the counting chamber.

Note: Remember to correct the results when a dilution is used, as the Corning®

Cell Counter does not correct for this automatically.

Counting Chamber Placement

The optical window of the Corning Cell Counter must be clean. You can check this by

monitoring the live screen. If any debris is present, you will see it in the CytoSMART™

Cloud App.

If any debris is visible, clean the optical window using lint-free wipes before placing the

counting chamber in the adapter plate.

When the sample is loaded in the counting chamber and the adapter plate has been

installed, you can place the counting chamber in the adapter plate that has been

inserted on the Corning Cell Counter.

4.0 Performing a Cell Count

After placing the loaded counting chamber on the Corning® Cell Counter, you can see an

image of the cell sample in the live screen (Figure 17).

Focusing of the Cells

Use the focus knob to adjust the focus of the image. To focus, turn the knob clockwise or

counter clockwise. For best results, ensure you have the correct focus on your tablet or PC.

For proper analysis, you should be seeing a contrast: living cells having bright centers

and dark edges (Figure 18). To achieve this, using the focus knob, set the focus just

above the cell.

We recommend trying multiple focus levels to determine which focus setting to use.

Figure 18. Correct focus settings

Figure 17. Live screen of the CytoSMART™

Cloud App

Important: The image seen on the live screen is a digital magnification of the field of

view, to allow for better focusing. The unmagnified image can be seen by clicking on the

magnifying glass (Figure 17A).

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CORNING CELL COUNTER Instruction Manual

Performing a Count

When the focus is set, you are ready to perform a count. To obtain your cell count, follow

the protocol below:

1. Make sure that the counting chamber is correctly placed in the adapter plate.

2. Check the image to see if the focus is correct – correct it if needed – and select

“Count” from the start screen (Figure 17).

3. The CytoSMART™ Cloud App will now analyse the image and present the results.

Within the analyzed image (Figure 19), you will see green and red circles (A). Green

circles represent living cells, red circles are dead cells. Next to the image, the number

of (live and dead) cells is displayed (B) together with experiment data (C).

4. In this window you can also add notes (D). These will be included in the CytoSMART™

Connect environment.

5. If you want to perform another cell count, simply press “Start new cell count” (E).

Figure 19. Overview of the result page

5.0 Connecting to the Cloud

One of the features of the Corning® Cell Counter is that the experiment results can be

accessed through the CytoSMART™ Connect. To access the cloud, the following items

are required:

 PC, tablet, or another device with a web browser.

 Active internet connection.

 Cloud login details.

After starting an experiment, the captured images will be stored in the cloud. To allow

for this, make sure the system has an active internet connection. All time logs in the

cloud are set to GMT.

Confirmation Email

After starting an experiment, a confirmation email is sent that contains a link to the

cloud (Figure 20).

Important: If you did not receive a confirmation email, please check your spam folder.

Alternatively, go to https://cloud.cytosmart.com/ to log in and view your projects and

experiments.

Figure 20. Confirmation email

18 19

CORNING CELL COUNTER Instruction Manual

This link will lead you to the CytoSMART™ Connect login page (Figure 21). Log in using

the credentials you created earlier. If you have multiple customer environments, choose

which environment you would like to access (Figure 22).

Figure 21. Connect login page Figure 22. Choose an environment

Figure 23. Project overview

Experiment Overview

After selecting a project (C), you will get access to an overview of all experiments

assigned to that project (Figure 24). At the top, you can see an overview of the project

data (F). As with the project overview, use the ‘Search’ bar (G) to find a specific

experiment within a project.

Figure 24. Overview of all experiments assigned

E D

Project Overview

After logging in to the Cloud, you will get access to an overview of all projects (Figure 23).

Navigating the portal is simple. If you have multiple projects in an account, use the

“Search Projects” bar (A) to quickly search for a project. To create a new project, select

“New Project” (B). This is similar to beginning a new project, as described in Section 3.

It is not possible to start an experiment from the cloud.

To access a project and all its assigned experiments, select the projects corresponding

“Project Icon”, “Project Title”, or “Arrow Icon” (C).

To delete a project, select the “Waste Bin Icon” (D). Select the “Pencil Icon” (E) to edit

project details. This allows you to change the project name and its description.

Select a specific experiment by clicking on its “Experiment Image”, “Experiment Title”, or

the “Arrow Icon” (H). This will bring up the experiment page of that specific experiment.

To edit details of an experiment, select the “Pencil Icon” (E). This allows you to change

the name and description of an experiment. To delete an experiment entirely, select the

“Waste Bin Icon” (D).

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