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RWD C100-Pro User Manual

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C100-Pro
Automated Cell Counter
User Manual
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Summary of Contents for RWD C100-Pro

  • Page 1 C100-Pro Automated Cell Counter User Manual...

  • Page 2: Intellectual Property Right

    RWD has the right to use the instruction as confidential information. Any individual and/or organization shall not disclose the instruction of all or part of the information by any means without RWD’s written permission. Nor shall any other person or organization be allowed to obtain all or part of the information of this instruction manual by any means.

  • Page 3: Table Of Contents

    CONTENTS INTRODUCTION ..........................1 ............................1 VERVIEW ....................... 1 OMPREHENSIVE ESCRIPTION ........................1 RODUCT EATURES ....................2 NVIRONMENTAL EQUIREMENTS ........................2 RODUCT ARAMETER ......................3 EFINED ARAMETER ..........................3 RODUCT SYSTEM SAFETY ..........................4 ..........................4 AFETY YMBOLS ........................5 AFETY NFORMATION 2.2.1 Electrical safety ...........................
  • Page 4 5.5.1 Language Switching ........................29 5.5.2 Replacement of Fluorescence cube ................... 29 5.5.3 Network setting .......................... 30 5.5.4 Server Configuration ......................... 31 5.5.5 Password change ........................33 5.5.6 Electronic record ........................33 5.5.7 User manage ..........................34 5.5.8 Date&Time ..........................35 5.5.9 Standby setting ..........................

  • Page 5: 1- Introduction

    1- Introduction 1.1 Overview Thank you first for choosing the C100-Pro Automated Cell Counter manufactured by the RWD Life Science Co., Ltd (hereinafter referred to as the RWD)! For better use of this product, please read the supplied instructions carefully before the initial installation and use of this product.

  • Page 6: Environmental Requirements

    Temperature: -20℃~60℃ Storage environment Humidity: 10%~93% (non-condensing) Altitude:＜2000m Choose the AC adapter supplied by RWD Input voltage: 90 V~264 V, 50/60 Hz, 2 A Working power supply Output voltage: 24V DC; output currency: 1.5 A Voltage and currency fluctuations should be within 5% of the working voltage.

  • Page 7: Self-Defined Parameter

    1.6 Self-Defined Parameter Parameter Description 0~255 Cell size 4~60μm Cell circularity 1~100 Cell brightness 0~255 Bright field luminance 0~100 Fluorescence cube luminance 0~100 Magnification (%) 100~800 Focus 0~2,400 Support 17 groups of custom Profiles, 1 group of system Profile options preset Profile 1.7 Product List Configuration...

  • Page 8: 2- System Safety

    2- System Safety 2.1 Safety Symbols Flammable environment hazard Never operate the instrument in an environment with flammable gases Electromagnetic interference hazard Make sure to operate the instrument in a controlled electromagnetic environment to avoid any risk associated with instrument failure. Radiation hazard Follow all applicable radiation safety procedures all the time when handling radioactive samples.

  • Page 9: Safety Information

    2.2 Safety Information 2.2.1 Electrical safety The following provisions should be followed for the safe use of the electrical devices accompanied with the instrument: a) Never disassemble the instrument without any authorization. b) Check whether the working voltage of all components are correct before connecting the instrument to the mains.

  • Page 10: General Safety Requirements

    In case the instrument breaks down or requires any service, shut down the instrument and contact with RWD’s after-sales personnel. 2.2.4 Biosafety It is possible that biological samples (such as samples of tissues, body fluids and blood of human and animals) may transmit infectious diseases.

  • Page 11: 3- Product Structure

    3- Product Structure  Automated Cell Counter Figure 3-1 Figure 3-2 1：Touch screen display: 7 inch capacitive screen. 2：USB port 1: the count data and image can be downloaded onto an external device with the USB port for storage. USB flash drive format: FAT32. 3：Counting slide port: for inserting the counting slide loaded with samples.
  • Page 12  Cell counting slide The plastic disposable closed counting slide consists of 2 chambers for loading samples, which can be used for loading the same sample as duplicates into 2 chambers for reproducibility validation. Cell count is performed in the center of each chamber. The total volume of cell for count is 20μL, which is equivalent to two small squares in a standard blood cell counter.

  • Page 13: 4- Preparation

    4- Preparation 4.1 Start-up When the instrument is properly wired, users are allowed to start up the cell counter. Press the power switch to start up the instrument and enter the main interface as shown in the picture below. Initial account: Admin Password: RWD2002 Figure 4-1 After entering the correct account and password, click "Login"...

  • Page 14: Profiles

    4.2 Profiles In the bright field (BF) assays, users are allowed to set the threshold values of the count results based on the profile sliders of size, brightness and circularity; in the fluorescence field (FL) assays, users are allowed to adjust the RFU slider as required to set the threshold values of the count results.

  • Page 15: Adjusting Parameter

    Fluorescence field (FL) Mode The following figure shows the adjustment of the fluorescence threshold values. The count result in this mode includes the total cell recognition chart, total cell concentration, AO/GFP, DAPI, PI cell concentration, total cell size/quantity distribution histogram, RFU/cell quantity distribution histogram, etc.
  • Page 16 to 4μm. When there are much debris in the sample, the interference of cell debris can be eliminated by adjusting the recognized size range and setting the threshold value. Brightness: drag the slider to enable the system to screen the cells by brightness while counting.
  • Page 17 After imaging, the device will automatically calculate the RFU of the background, and set the lower limit of RFU above the background, so as to filter it. The automatically calculated background can usually be distinguished from the fluorescent cells, but its threshold value will fail in the event of too strong the fluorescence background or too weak cell fluorescence, possibly leading to the removal of the weak cell fluorescence or the presence of residual background fluorescence.

  • Page 18: Load Profiles

    focusing results will be presented first, and then the count results will be output after manually clicking the [Count] key. The quick mode is suitable for cell samples with no debris and moderate concentration. It can automatically focus and count after inserting the counting slide, which can greatly reduce the counting time.

  • Page 19: Sample Preparation

    In the left column of the Profiles interface, you can delete/ add/ rename/ Lock profiles by clicking the icons of respectively. Note: Locked parameters cannot be edited. Click lock key to lock the selected profile, the icon is also displayed on the right of the profile name;...

  • Page 20: Methods Of Staining

    4.5 Methods of Staining Tips: If you only need to count the total cell concentration, you could directly load sample without staining. 4.5.1 Ttrypan blue staining Recommended staining method: 1) Mix 10μL cell suspension with 10μL Trypan blue solution. 2) After pipette and mixing well, transfer 10μL of stained cells into the cell counting slide for analysis.

  • Page 21: Loading Samples Into Slide

    volume to kill the cells for staining if don't need to count the cell viability. 4.6 Loading Samples into Slide 1) After pipette and mixing well, transfer 10μL of sample into the cell counting slide for analysis. 2) Allow the sample to stand for 30 seconds in the slide, and insert the sample-loaded slide into the slide port.

  • Page 22: 5- Cell Count And Cell Survival Rate Assays

    5- Cell Count and Cell Survival Rate Assays If the system recognizes insertion of the counting slide, the file name editing interface will pop up. Shown as figure 5-1. Figure 5-1 Enter the file name and click [Enter] or click [skip] (“skip” means that the default name or last typed name is selected) to enter the count interface.
  • Page 23 For setting the dilution factor of the sample. Click “1×” to pop up a numeric keypad. The range of setting value: 1.00~999.00. Note: The default dilution of Trypan Blue mode is “2×”, and the default dilution of "AO+PI" mode is “2×”. Dilution factor of sample: The cell concentration counted by cell counter is the actual concentration of the cells on the current counting slide, which may be obtained by dilution of the original solution, so you need...

  • Page 24: Focusing

    Figure 5-3 A Overexposure: the light source is too bright, and the background is white, and the cell details are covered by light, which affects cell recognition. B Underexposure: the light source is too low, and the background is too dark, and the cells as a whole are too dark, which affects the judgment of the live and the dead.
  • Page 25 Figure 5-4 A Normal focus: well-outlined cells, with a clear aperture in the middle of live cells, and opaque dead cells due to trypan blue staining. B Abnormal focus: blurred cell contour, presented as opaque dead cells. Two possible results from incorrect focusing: ...
  • Page 26 Figure 5-5 Click to pop up a histogram, as shown in the figure below, and click it again to exit. Figure 5-6 Click to show cell marking options, as shown in Trypan Blue mode. Click the green circle to mark the live cell, then all the live cells will be indicated by the green circle, shown as figure 5-7.
  • Page 27 Figure 5-7 Click to enter the Adjustment interface to re-adjust the Profiles. Click to enter the Dilution Calculator interface, as shown in the figure below, where users are allowed to calculate the volume of existing cell stock solution and the volume of buffer solution to be mixed and diluted to the target volume and concentration.

  • Page 28: Fluorescence Assays

    5.2 Fluorescence Assays The following figure shows the data acquisition interface in FL assays. This interface allows you to perform such actions as zooming in, fine tuning of focus and light source brightness adjustment, so as to capture optimized images for accurate count of cell concentration or dead/ live cells.
  • Page 29 Figure 5-10 A. Fluorescence overexposure: brighter fluorescent light source and too strong fluorescence background, affecting cell recognition and observation. B. Underexposure of fluorescence: dimmer fluorescent light source, black overall fluorescence of the cells, darker overall fluorescence of the cells, and some cells with lighter staining missing in counting.
  • Page 30 Results interface of [Channel] Figure 5-11 Results page of [Application] Figure 5-12 is a for channel selection ; the channel menu will pop up when it is clicked. The channel menu is only displayed in the FL assays. Users can switch the light source channel to check the fluorescence status of fluorescent cells under different light source states.

  • Page 31: Results Storage

    histogram in the BF assays or FL assays, of which the histogram in the BF assays is in cell size- based distribution, while the histogram in the FL assays is in RFU intensity-based distribution. Figure 5-13 Refer to the section BF assays for relevant function keys. 5.3 Results Storage After count, you can choose to save/ export the original image, cell marking image, count parameters, cell count report and other data.

  • Page 32: Previous Counts

    Figure 5-14 5.4 Previous Counts Click [Previous Counts] on the main interface to enter the following page, where you can view the pevious data of the instrument,you can delete, export, and view data. Figure 5-15 Click the drop-down arrow at to select an account and view the historical data of the corresponding.

  • Page 33: Language Switching

    Figure 5-17 5.5.1 Language Switching For setting the system language to “ Chinese ”/ “English” Figure 5-18 5.5.2 Replacement of Fluorescence cube 1) First click the [Reset FL Cube] on the [System Setting] interface to move the fluorescence cube up. After the reset is successful, please replace the fluorescence cube when turns off the power switch and disconnects the device power.

  • Page 34: Network Setting

    Figure 5-19 4) Take out the screwdriver embedded in the back panel. 5) Select the fluorescence cube to be replaced, loosen screw 1 and screw 2 on one of the fluorescence cubes counterclockwise, then screw the screwdriver clockwise into hole 3 in the centre of the fluorescence cube until it stops rotating, and then pull out the fluorescence cube and screwdriver horizontally.

  • Page 35: Server Configuration

    Figure 5-21 5.5.4 Server Configuration The instrument can be connected to the server by inputting the same IP address and port number as the PC server to get the real-time data uploading. It is necessary to set it on both the operation interface and computer.
  • Page 36 Then tap on the device main interface. Click “System Setting” →“Server” and then enter the “Server setting” page, as shown in Figure 5-23. Figure 5-23 Type the correct IP address and port and then click [Save], the successful connection status is shown as figure 5-24.

  • Page 37: Password Change

    5.5.5 Password change Click to enter the [Password Change] page, enter the old password, enter the new password twice, and click [OK]. Figure 5-25 5.5.6 Electronic record [Electronic record] displays the user account, operation time, operation type and operation details of the device. Filter and view the information according to the account name and time range.

  • Page 38: User Manage

    5.5.7 User manage On the “User manage” page, you can add, delete, edit, disable or enable accounts. Note: Only the account “Admin” can configure an account, and has all permissions of the device. Figure 5-27 Click [Add] to enter the “Add user” page, as shown in Figure 5-28. Here you can configure user account permissions, user level, account name, and so on.

  • Page 39: Date&Time

    Figure 5-29 5.5.8 Date&Time Only the account “Admin” has the permission to edit the date e& time of the system. Enter the password as prompted to go to the date & time setting page. Click the value of "Date Format" and "Time Format" to switch different modes of date & time. Click "+"...

  • Page 40: Software Update

    Figure 5-31 5.5.10 Software Update The latest version of the software is available for updating to the instrument by the USB flash drive. Method: First copy the latest software to the USB flash drive, insert the USB flash drive into the USB port, when the USB flash drive is successfully recognized,you could click [Update Now] When the software program in the USB flash drive is successfully recognized, the keyboard...

  • Page 41: 6- Maintenance

    6- Maintenance 6.1 Cleaning  Cleaning the touch screen 1) Gently wipe off the touch screen with a soft lint free cloth moistened with an LCD cleansing detergent. Be gentle and cautious during cleaning. Wipe the touch screen dry immediately after cleaning. 2) Prevent ingress of cleaning solvent into the Power button, power socket, counting slide port or USB port.

  • Page 42: 7- Fault/Alarm

    7- Fault/Alarm This section introduces the problems (faults) that are common to the product and their possible causes and solutions. Fault/Alarm Solution Ensure that there are no bubbles or debris Autofocus fails to focus cells well interfering with the autofocus within the sample in the current view.

  • Page 43: 8- Warranty

    If the reworked equipment was found to have been unauthorised disassembly, RWD will not provide after-sales service such as warranty, free maintenance and parts replacement.
  • Page 44 RWD Life Science Web: www.rwdstco.com Add: 850 New Burton Road, Suite 201, Dover, DE 19904, Kent, Delaware, USA Add: 19F, Building 9A, Vanke Cloud City III, Liuxin 4 Street, Nanshan District, Shenzhen518000, Guangdong, P.R. China Tel: +001-858-900-6602 +86-755-86111286 After-sales Service: +86-755-86111281...

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