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RWD C200FL User Manual

Fluorescent automated cell counter
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C200FL
Fluorescent Automated
Cell Counter
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Summary of Contents for RWD C200FL

  • Page 1 C200FL Fluorescent Automated Cell Counter User Manual...

  • Page 2: Intellectual Property Right

    RWD’s registered trademark or mark, these trademarks and the related security mark belong to RWD’s intangible property. The use of non RWD’s trademark or mark in this instruction manual are only for editing purposes, without other purposes, the rights belong to their respective rights owners.

  • Page 3: Table Of Contents

    CONTENTS PREFACE ............................... 1 OVERVIEW ........................... 1 ..........................1 DVANTAGES ..............2 EQUIREMENTS FOR INSTRUMENT ENVIRONMENT ......................2 RODUCT PARAMETERS .......................... 3 RODUCT LIST .......................... 3 NTENDED USE IMPORTANT INFORMATION AND SECURITY ..............4 ........................4 AFETY SYMBOLS ........................... 4 AFETY 2.2.1 General safety ........................
  • Page 4 ........................32 ROWTH CURVE ......................... 33 ELETE .......................... 34 XPORT DATA SET ..............................35 ........................... 36 OUNT TYPE ..........................36 8.2.1 Add/Copy/Delete/Edit Cell Type ..................38 ........................40 OCUS ETTING ..................... 41 NSTRUMENT CALIBRATION 8.4.1 Calibrate setting ........................ 43 8.4.2 Calibrate data ........................

  • Page 5: Preface

    Preface First of all, thank you for choosing the fluorescent automated cell counter RWD! Before installing and using this product, make sure that you read all the attached schedules carefully for better future reference. RWD is committed to continuously improving product functions and service quality, and reserves the right to change any products described in this user manual and the contents of this user manual without prior notice.

  • Page 6: Requirements For Instrument Environment

     High-performance fluorescent components, clearer fluorescent imaging  App type counting program, grouped management, and one-click call  6-channel slide, one-click automatic sample feeding  500 G storage, one-click export of concentration/viability/agglomeration rate/growth curve  The system design complies with FDA 21CFR PART11 1.2 Requirements for instrument environment Please prepare the operating environment as the conditions listed in the table below to ensure the operability and safety of the device.

  • Page 7: Product List

    Trypan blue dye used 1.5 Intended use The intelligent fluorescent automated cell counter C200FL manufactured by RWD is a cell analysis system that combines intelligent image recognition and advanced optical imaging technology for accurate counting. The self-developed intelligent algorithm can effectively recognize live and dead cells, and achieve 5s rapid imaging and accurate counting.

  • Page 8: 2- Important Information And Security

    2- Important information and security 2.1 Safety symbols Hazards in flammable environment Do not operate the device in the presence of flammable gases. Electromagnetic interference hazards To avoid the danger of device fault, always operate the device in a controlled electromagnetic environment.

  • Page 9: Electrical Safety

    RWD. Any changes not authorized by the manufacturer may cause operational failure of the instrument. 2.2.2 Electrical safety In order to safely use the built-in electrical appliances, the following provisions shall be...

  • Page 10: Biosafety

    d) Minimize the risks of inhaled chemicals. Do not open containers for chemicals. Use chemicals only in well-ventilated environments (e.g. fume hoods). Refer to the SDS for detailed safety guidance. e) Handle chemical waste in a well-ventilated environment. After emptying the waste container, seal the container with the included cover. g) Waste trays and bottles should be discarded in accordance with good laboratory practices and local environmental and health regulations.

  • Page 11: 3- Introduction To Product Structure

    3- Introduction to product structure 3.1 Cell counter main unit Figure 3-1...
  • Page 12 Figure 3-2...
  • Page 13 Figure 3-3 Component/interface Description Turn on/off (long press for off) cell counting system Status indicator: System switch button  Orange - Device powered up, counting system not (with status indicator) turned on  Green - Counting system in operation Slide socket To insert slide Power switch Device power on/off...

  • Page 14: Cell Counting Slide

    3.2 Cell counting slide Note: For best results, do not touch the optical surfaces of the counting slide and wear gloves when handling samples. Note: Reuse of disposable slides may affect count results. Figure 3-4...

  • Page 15: 4- Preparation Before Operation

    4.3 Slide loading 1) Pipette and mix the sample with a pipette, and load the sample into the chamber of the slide. 2) Allow the sample mixture to stand in the slide for 30s. Note: Please use the supplied RWD counting slide.

  • Page 16: 5- Startup And Login

    5- Startup and login When the device is properly connected, press the power switch to power on the device, and then click the system switch button to turn on the cell counter. After the system initialization is completed, automatically enter the login page, as shown in Figure 5-1. Click “...

  • Page 17: 6- Counting

    6- Counting Figure 6-1 6.1 Fast counting As shown in Figure 6-1, commonly used count types are preset in the [Count] interface: Total cell count, Trypan blue, Trypan blue-Auto Exp, AOPI, GFP, Faint-GFP, RFP, Faint-RFP. Select the count type, a prompt of “Please insert slide” will pop-up, as shown in Figure 6-2. Figure 6-2 Insert the slide according to the prompt, and the interface of “Edit Counting Information”...
  • Page 18 Figure 6-3 The number of the sample chamber on the slide corresponds to the number of the interface channel, and the slide is inserted as indicated by the red arrow. Figure 6-4 1) Experiment Name/Sample Name: User-defined, 0-20 characters. 2) Cell Type: Click the drop-down arrow to select all preset cell types under this count type. To add cell type, refer to Section 8.2.

  • Page 19: Count Results

    Figure 6-5 6.2 Count results Based on the count type selected, the count results will present different channel combinations. If AOPI count type is selected, five channels of BF, AO, PI, AO+PI and Merge will be displayed. Figure 6-6 Description Switch between different sample chambers Display the general informations Observe the detailed parameters...
  • Page 20 Switch between different views Switch between different channels Count results of current view & overall Switch original/marker view Common count types correspond to the following channels: Total cell count Trypan blue Trypan blue-Auto Exp AOPI Faint-GFP Faint-RFP Count results - Overall Cell viability = sum of live cell number in all views/sum of total cell number in all views.

  • Page 21: Cell Marker

    6.2.1 Cell marker Marker enabled Marker disabled Figure 6-7 6.2.2 Image zoom/restore/move Image zoom: Use pinching gestures to zoom in and out of the image. Zoom in/out: Double-click the image area to zoom in/out the image. Move: When zooming in, long press to drag the image display area. 6.2.3 View parameters Click [View] to enter the interface as shown in Figures 6-8.

  • Page 22: Parameters Adjust

    Main parameter items and their definitions: Parameters Meaning BF channel parameters, minimum cell size. Cells smaller than this diameter will be MinSize deleted. BF channel parameters, maximum cell size. Cells bigger than this diameter will be MaxSize deleted. BF channel parameters, cell viability threshold. If the viability of a single cell is Alive/Dead lower than the threshold, it will be judged as dead cell, and if it is higher than the threshold, it will be judged as live cell.
  • Page 23 Parameters adjustment Figure 6-9 ： Note [More] is only available in BF channels of BF type and fluorescence type, as shown in Figure 6-10. Figure 6-10 Click to switch to other channels (such as AO channel) for parameters adjustment, as shown in Figure 6-11.
  • Page 24 Figure 6-11 to enter the “Help” pop-up window as shown in Figure 6-12 to view the Click on the help relevant interpretation of parameters. Figure 6-12 Main parameter items and setting range: Parameters Setting range 3.00-180.00 μm MinSize 3.00-180.00 μm MaxSize Alive/Dead 0-255.0...

  • Page 25: Re-Analysis

    MaxArea 1-9999 FlouTH 1-255 6.2.5 Re-analysis The user can re-select the parameters of other cell types from the saved data for re-analysis, and the re-analyzed data will be saved as new data without overwriting the original data, and can be traced back to the original data through the unique sample ID in the remark column at the end of the “Data”...

  • Page 26: Calculator

    Figure 6-15 RFU distribution-AO 6.2.7 Calculator Click [Calculator] to enter the interface as shown in Figure 6-16. [Calculator] is used to calculate how much volume of existing cell solution needs to be mixed with buffer when diluted to the target volume and target concentration. Manually enter the target concentration and target volume, and the system will automatically suggest values.

  • Page 27: Add Count Type

    6.3 Add count type Count type needs to be added from external device, and make sure the external device and the counter are connected before clicking key to add the count type. When the system recognizes that there is a valid count type in the external device (for example, the AOPI-1 type exists on an external device and is identified), the password entry interface will pop up, and enter password of current user, and the system will automatically enter the interface as shown in Figure 6-17.

  • Page 28: Copy/ Delete/Export/Edit Count Type And Icon Groups

    6.4 Copy/ delete/export/edit count type and icon groups As shown in Figure 6-19, long press the icon to copy/delete/export/edit count types. Note: The preset count types can only be copied, not be edited, deleted or exported. Examples of figures (a) and (b) (a) Long press the preset type icon (b) Long press the custom type icon Figure 6-19...
  • Page 29  Copy Click [Copy] to enter the interface as shown in Figure 6-21. Figure 6-21 The user can customize the name of the count type, number of views, cell type and dilution factor after copying. Click [Confirm] to complete the setting. ...
  • Page 30 Figure 6-24 Select the chamber, click [Confirm] to enter the interface “Edit Count Type”, and set the general information and exposure parameters of the count type, as shown in Figure 6-25. Click [Save] to complete editing. Note: The content displayed on the interface varies according to the count type. Edit General Information: Figure 6-25...
  • Page 31 The parameter adjustment range is shown in the following table: Parameters Description Type Name Custom, enter 1-20 characters Number of views Options from 3, 2 and 1 Options from 1.00X, 2.00X, 10.00X, and 100.00X). Dilution Factor Custom entry range: 1.00 ~ 10000.00 Allowable CV value of total concentration 0-100%, resolution 1% 0-100%, resolution 1%.
  • Page 32 Figure 6-26 Edit exposure parameters: The exposure parameter is shown in Figure 6-27. You can set the exposure parameters for each channel (such as BF, FL1, FL2). Click to select the channel and move the slider or click to adjust the Exposure and Gain. When the exposure parameters are adjusted to the best image display, click “Save”...
  • Page 33 The parameter adjustment range is shown in the following table: Parameters Description Exposure of BF 0~72 Exposure of fluorescence 0~592809 Gain 0~48.0 Note: When the “Trypan Blue - Auto Exp” count type is selected, auto exposure will be performed. If the brightness of the picture is moderate, the exposure of BF need not to be adjusted. Since the application of fluorescence is complex, the result of auto FL exposure is affected by the RFU and the quantity of fluorescent cells.
  • Page 34 Figure 6-28 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 35: 7- Data

    7- Data Figure 7-1 The interface “Data” displays all experiment data under the current user by default. Drag the slider left and right to view more data. Click the drop-down arrow of Account to switch to display all accounts and experiment data that the current user has access to.

  • Page 36: Growth Curve

    has a set value.) ② When “Instrument Calibration” is on, and instrument calibration is not performed or calibration is expired, the count result is Fail. (If “Instrument Calibration” is turned off, this item is not judged.) ③ Count results for each chamber: The concentration/viability bias among the 3 views exceeds the upper limit of the allowable CV value of total concentration/viability (if any)/transfection rate (if any) as set in Count type-General (Figure 6-24).

  • Page 37: Delete Data

    Figure 7-4 By default, X-axis is displayed according to the “Sequence” arrangement. Click the drop-down arrow to switch to the “Time” order display. The drop-down options on the Y-axis are related to the current count type, for example, when using the total cell count type, the drop-down options show the total concentration, total cell number, average size, and agglomeration rate.

  • Page 38: Export Data

    7.3 Export data To export data to an external storage device, make sure you insert a USB flash drive in the correct format (FAT32/NTFS/EXFAT). After checking the result data to be exported in [Data], click [Export] to enter the interface as shown in Figure 7-6.

  • Page 39: 8- Set

    8- Set Click [Set] to enter the setting interface. The Admin account setting interface is shown in Figure 8-1, and the non-Admin account setting interface in Figure 8-2. Figure 8-1 Figure 8-2...

  • Page 40: Count Type

    8.1 Count type Click [Count type] to enter the interface as shown in Figure 8-3. Default display system preset count type. Please refer to Sections 6.3 and 6.4 for descriptions of Add/Copy/Delete/Export/Edit count types. Multiple or all count types can be checked for the above operations. Preset count types cannot be checked/All selected.
  • Page 41 Figure 8-4 Click [Details] to view the details of the current cell type, refer to Figure 8-5. Figure 8-5 For the cell type under user-defined count type, add, copy and edit can be performed after blue shading is selected. Such as switch the count type to the “AOPI-1” and select the cell type name of “293”, shown in Figure 8-6.

  • Page 42: Add/Copy/Delete/Edit Cell Type

    Figure 8-6 Selection recommendations for cell type: 1) [Small_Cell] is recommended for immune cells smaller than 10 μm, such as mouse spleen cells. 2) [293] or [CHO] can be selected for conventional cell lines larger than 10 μm. 3) Please select [PBMC] for human PBMC. 8.2.1 Add/Copy/Delete/Edit Cell Type ...
  • Page 43 The default cell type name is “cell”, which can be customized. Click to switch the channel and enter the corresponding parameters. Click [Edit (View)] to enter the interface as shown in Figure 8-8, select the chamber where the target sample has been added and click [Confirm] to enter the counting state. Note: It is needed to insert the slide and ensure that the sample that requires cell parameter adjustment has been added into the chamber.

  • Page 44: Focus Setting

     Copy Check one of the cell type data and click [Copy] to enter the interface as shown in Figure 8-10. Customize the cell type name and click [Confirm] to complete the copy. Figure 8-10  Delete Check the cell type data to be deleted, and click [Delete] to enter the interface as shown in Figure 8-11.

  • Page 45: Instrument Calibration

    Figure 8-12 8.4 Instrument calibration The “Instrument Calibration” function is used to calibrate cell concentration, viability, and cell size, as shown in Figure 8-13. Only the Admin account can perform instrument calibration. Figure 8-13 Select the item to be calibrated, such as “Viability-BF”, click the corresponding [Cali] button, insert the slide according to the screen tip, and then automatically enter the “Edit Calibration Information”...
  • Page 46 Figure 8-14 Note: Before starting calibration, make sure that the same standard has been loaded into all chambers of this plate and matches the calibration type and standard value selected, or calibration will fail. Display calibration results after calibration is complete Figure 8-15...

  • Page 47: Calibrate Setting

    8.4.1 Calibrate setting Click [Calibrate Setting] to enter the interface as shown in Figure 8-16, and set the calibration parameters of “Concentration/Viability-BF/Viability-FL/Cell Size”. Figure 8-16 Parameters Description Expiration Date 1-365 Standard concentration 1-3 1.00 × 10 -9.99 × 10 (concentration) Standard viability 1-3 (viability-BF, 0-100% viability-FL)

  • Page 48: Calibrate Data

    “CV value” in [Calibrate setting] is the CV value of 6 chambers corresponding to Concentration, Viability-BF, Viability-FL and Cell size. 2) Calibration setting - Allowable bias of viability a) Overall viability of chamber N = sum of live cell number in 3 views/sum of total cell number in 3 views.
  • Page 49 Figure 8-17 Click [Chamber Information] to view the results information of all chambers of this data (the display parameters depend on the calibration type), as shown in Figure 8-18. Drag the slider bar left or right to browse more parameters. Figure 8-18 The format of exported calibration data is as follows.

  • Page 50: Electronic Records

    8.5 Electronic Records The interface [Electronic Records] displays the account, operation time, operation type and operation details of this instrument, which can be filtered and viewed according to the account name and operation time range. Click [Export] to select the export path as external storage device or server, and follow the prompts.
  • Page 51 Note: Account can only be configured by the Admin account, and has all authorizations of this instrument. Note: Deleting an account will delete the count type, cell type, historical data, etc. corresponding to the user. Please use this function carefully. Click [Add] to enter the interface [Add Account] as shown in Figure 8-22.

  • Page 52: Change Password

    Click [Confirm] to complete the authorization for the account. Figure 8-24 8.7 Change password Click to enter the interface [Change Password], enter the current password, enter the new password twice, and click [Save]. Figure 8-25...

  • Page 53: Backup&Restore

    8.8 Backup&restore Click [Reset to Factory] and enter the Admin password according to the pop-up window prompt. After the system verification passes, all sets of the device will be restored to factory settings and all user data will be cleared. Please use this function carefully. If system backup is required, please ensure that the instrument has been connected to the external storage device, click [System Backup], enter the Admin password according to the pop-up window prompt, and export the count type, cell type, counting data, calibration data,...

  • Page 54: Corporate Information

    8.9 Corporate information Figure 8-27 Click [Corporate Information] to enter the interface as shown in Figure 8-27. User can customize the enterprise name: Click the input box to modify the enterprise name. If you want to change the interface display image, please first make sure that the instrument has to enter the interface “Change Image”, as been connected to the external device.

  • Page 55: Language Setting

    8.10 Language setting Click to switch system language, as shown in Figure 8-29. Figure 8-29 8.11 Brightness setting As shown in Figure 8-30, drag the slider bar to increase/decrease the screen brightness. Adjustable range: 1-100%. Figure 8-30 8.12 Network setting Network connection: Wired network and WLAN connections are available.

  • Page 56: Manual Setting

     Auto-acquisition Figure 8-31  Manual Setting Figure 8-32...

  • Page 57: Wi-Fi Connection

    8.12.2 Wi-Fi connection Click on “Wi-Fi connection”, then manually select the network account name and enter the password. Figure 8-33 8.13 Server setting Click [Server] to enter the interface as shown in Figure 8-34. The interface displays the current server connection status. Real-time data upload can be enabled/disabled by click , and when enabled, the system will automatically upload the data to the server in the selected format after counting finished.

  • Page 58: Date&Time

    8.14 Date&time Only administrator accounts can modify the date/time of the system. Enter the login password according to the prompt and enter the interface as shown in Figure 8-35. Figure 8-35 Optional date format: YYYY/MM/DD, MM/DD/YYYY, DD/MM/YYYY Optional time format: 24-hour and 12-hour system 8.15 Standby setting...

  • Page 59: Storage Space

    8.16 Storage space Click [Storage] to view the storage status of the current device. Display the total device storage space, available space, and used space. Figure 8-37 8.17 Manual Click [Manual] to get the user manual, please follow the manual instructions.. Figure 8-38 8.18 Software information...

  • Page 60: Local Upgrade

    Upgrade the latest software version of fluorescent automated cell counter to the instrument by a USB flash drive. First, copy the latest software upgrade package (connect the RWD aftersales) to the USB flash drive, insert the USB flash drive into the USB port. When the system successfully recognizes the USB flash drive, click [Local Upgrade].

  • Page 61: Maintenance

    8.20 Maintenance For after-sales maintenance personnel only. 8.21 Shutdown In the running status, long press the system switch 3 s to enter the interface as shown in the figure below. Click [Shutdown] to shut down the system. Note: Please remove the counting slide before shutdown, otherwise it may cause damage to the machine.

  • Page 62: 9- Alarm Information

    9- Alarm information Refer to the table below for handling alarm list. Note: When an alarm list appears, counting will be disabled, and the current count results will be abnormal and cannot be used as correct data. Error Alarm Alarm information Possible cause Solution code...

  • Page 63: 10- Troubleshooting

    10- Troubleshooting This chapter describes the possible failures encountered when operating the product, the possible causes of the failure, and the measures to be taken. Failure phenomenon Troubleshooting After the program upgrade, restore to factory settings. Count results cannot be saved Back up the data before restoring to factory settings.

  • Page 64: 11- Maintenance

    11- Maintenance 11.1 Product cleaning  Cleaning of touch screen 1) Wipe the touch screen of the instrument with a soft lint-free cloth soaked in LCD cleaning solution. Do not use excessive force during cleaning. Dry the touch screen immediately after cleaning. 2) Ensure that the cleaning solution does not enter the power button, power outlet, slide interface, or USB port.

  • Page 65: 12- Warranty

    If the repaired instruments are found to have been disassembled without authorization of RWD upon arrival, the Company will not provide after-sales services such as warranty, free maintenance and parts replacement. This warranty statement (including its limitations) is issued exclusively by RWD and covers...
  • Page 66 RWD Life Science Co., Ltd. Add: 10410 Corporate Drive, Sugar Land, TX 77478, USA Add: (Floor 9, 19&20 Building 7A, Floor 9 Building 7D) Room 1901, Building 7A, International Innovation Valley, Dashi 1st Road, Xili Community, Nanshan District, Shenzhen 518000, Guangdong, P. R. China Web: www.rwdstco.com...

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