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NanoEnTek ADAMII-CD34 User Manual

Bench-top fluorescence cell counter
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-CD34
Bench-top fluorescence cell counter
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Summary of Contents for NanoEnTek ADAMII-CD34

  • Page 1 -CD34 Bench-top fluorescence cell counter User manual...
  • Page 2 Every effort has been made to ensure that all the information contained in the ADAMII™ User Manual is correct at the time of printing. However, NanoEntek Inc. reserves the right to make any changes necessary without notice as part of ongoing product development.

  • Page 3: Table Of Contents

    Troubleshooting ............. 50 ~ 51 Error code ............... 52 ~ 53 Warranty ................. 54 Safety precautions ............55 Consignes de securite ........... 56 Safety symbols ............... 57 Product specifications ..........58 Ordering information ............. 59 Technical support ............60 ADAMII™ © NanoEntek, Inc.

  • Page 4: Introduction

    Introduction ADAMII-CD34 is an image-based bench-top cell counter that quantifies CD34+ cells. ADAMII-CD34 is a FDA 510(k) cleared instrument that can be used for mobilized peripheral blood (MPB) and hematopoietic progenitor cells apheresis (HPC-A) samples. ADAMII-CD34 is a CE approved instrument that can be used for mobilized peripheral blood (MPB) and hematopoietic progenitor cells apheresis (HPC-A) samples.

  • Page 5: Product Components

    Product components ADAMII CD34 consists of the following components. If any of the components is missing or damaged, please contact your local sale representative or send an email to ivdst@nanoentek.com. ADAMII™ CD34 Power cord with 4 adaptor cords USB connection cable (for U.S./Canada/ Taiwan/Japan, Europe or UK)

  • Page 6: Product Description

    Product description Front view State LED State LED Blue LED lights up when the instrument is connected to the laptop and the ADAMII-CD34 software is running. Slide holder Slide door Slide holder Where ADAMII-CD34 Assay Slide is inserted to. Where one can access the Slide holder.
  • Page 7 Power Power switch USB 3.0 port supply port Power supply port To connect the power adapter To turn power On and Off Power switch To connect USB cable to communicate USB 3.0 port with the laptop ADAMII™ © NanoEntek, Inc.

  • Page 8: Installation

    Connect cables 1. Plug the power cord into an electrical outlet and plug the DC output jack to the power supply port. 2. Connect USB 3.0 cable to the USB ports in the ADAMII-CD34 and the laptop. USB 3.0 port...
  • Page 9 2. Double-click ‘ADAM’ icon on the laptop to start the ADAMII-CD34 CD34 software software. ☞ Note: When it is turned on, the ADAMII-CD34 will be ready to use after a quick initialization. ☞ Note: When the ADAMII-CD34 is connected to the laptop and the ADAMII-CD34 software is running, the state LED will light up.

  • Page 10: Home Menu

    Home menu Log in At the ‘Home’ screen, enter the user ID and password to log in. Default ID and password are both ‘master’. User Click ‘User management’ when you need to manage accounts (e.g. to add a new user or to change password). To create a new ID, management fill out the blanks (user name, user ID, password, confirm PW) and click ‘Add’.
  • Page 11 1. Enter the User ID and set the date that you want to search. 2. Click ‘Search’ button. 3. The record of the selected user is displayed. ☞ Note: Only the master user can access the ‘’Audit trail’ . ADAMII™ © NanoEntek, Inc.

  • Page 12: Calibration

    Calibration Calibration 1. After turning on the ADAM II CD34 first time, the instrument needs to be calibrated. Subsequently, it is recommended that the calibration procedure is done regularly or at least once a week. A pop-up window for calibration appears as shown below.
  • Page 13 5 sec. [Shake bottle vigorously] [Vortexing] ☞ Note: ADAMII Calibration Beads are included in ADAMII-CD34 kit. Before use, the ADAMII Assay Slide should be removed from refrigerator and kept at room temperature for at least 30 minutes. The reagents should be kept at 4ºC.
  • Page 14 Calibration Calibration CAUTION: procedure • When loading samples to the slide, please release sample gently and slowly to avoid bubbles because bubbles interfere image analysis. Warning: • Calibration Beads loading error: - Ensure that calibration bead solution is fully loaded into slide. - If loading volume is not enough, calibration can be inaccurate.
  • Page 15 7. Enter lot number of Calibration Beads. Calibration procedure CAUTION: • Lot number has to be entered after inserting slide. 8. Select ‘Bright’ channel and adjust the focus using the focus buttons. To view beads 2x bigger, press ‘Zoom in’ button. ADAMII™ © NanoEntek, Inc.
  • Page 16 Calibration 9. Press the ‘Save BR Focus’ after adjusting focus. Calibration procedure ☞ Note: Please check ‘ADAMII™ Focus guide’ before setting the focus. [Focusing example - representative images] * Enlarged image Bright- Good Focus Bright- Bad Focus * Enlarged image FL- Bad Focus FL- Good Focus •...
  • Page 17 11. Press the ‘Save FL Focus’ after adjusting focus. ☞ Note: After saving the focuses of “Bright” and “FL3”, the focuses of the other channels (FL2 and FL1) will be automatically adjusted. 12. Press the ‘Run’ button. 3~4 min ADAMII™ © NanoEntek, Inc.
  • Page 18 Calibration Calibration 13. When the calibration is completed, the results will be displayed. procedure ☞ Note: If any of the results are “FAIL”, please contact the manufacturer or distributor. ☞ Note: If any of the results are “Low”, please check the focus or expiration date of the Calibration Beads.
  • Page 19 Calibration Calibration 15. The calibration results are displayed. To save the results, click ‘Save’ button. procedure 16. Press “Close” button to finish calibration. ADAMII™ © NanoEntek, Inc.

  • Page 20: Measurement

    Measurement Sample Prepare sample following the instruction enclosed in ADAMII-CD34 kit. preparation To measure a sample, click ‘Measure’ menu. Load sample Load 25 uL stained sample into an ADAMII Assay Slide. [Load 25μL sample] ☞ Note: Please refer to page 14 to check cautions for loading correct volume .
  • Page 21 2. Press the black bar to open the slide holder. 3. Insert the Assay Slide into the slide holder. ☞ Note: Assay Slide should be inserted all the way in. Please see the instruction in the next page. ADAMII™ © NanoEntek, Inc.
  • Page 22 Inserted in opposite direction Inserted upside down (See the arrow mark) 4. Close the slide holder by pressing down the top portion of the holder tightly until it ‘clicks’. Warning: If the slide holder is not properly closed, ADAMII-CD34 will give a warning.
  • Page 23 5. Close the slide door. assay slide 6. Select sample type, dilution factor and the number of frames. 7. Press the “Insert sample’ button on the ADAMII-CD34 software. The slide holder containing the Assay Slide will move into the measuring position.
  • Page 24 Measurement Measure menu The functions of each part in the ‘Measure’ menu are as follows. - Initial mode 1. Counting mode 2. Display area 3. Detection channel 4. Insert sample 5. Counting results area 6. Test information...
  • Page 25 (the dilution effect during sample preparation has been set in the Setting). • Frames: ADAMII-CD34 can take up to 75 frames of images per sample. • Viability: Check the circle to include viability measurement. ② Display area Live feed of camera will be shown in this window.
  • Page 26 Measurement Measure menu After completing image acquisition and analysis, the results will be displayed here. (Refer to page 33) - initial mode ⑤ Counting result area ⑥ Test Detailed information of the sample will be displayed here. information...
  • Page 27 Measurement Measure menu Once the Assay Slide is inserted, focusing mode will appear. - Focusing mode 1. Display area 2. Channel 3. Focus option 4. Save focus 5. Run sample 6. Eject ADAMII™ © NanoEntek, Inc.
  • Page 28 Measurement Measure menu Live feed of camera will be shown in this window. - Focusing mode ① Display area ② Channel Indicate the channel that is being displayed. • CD45 • CD34 • Viability for live/dead marker ☞ Note: Viability channel will be measured only when the viability circle is selected.
  • Page 29 2 fluorescence channels will be automatically set based on the CD45 focus position. Auto focus : If you press the “Auto focus” button, the focusing will be adjusted automatically. [Save BR focus] [Save FL3 focus] ADAMII™ © NanoEntek, Inc.
  • Page 30 Measurement Measure menu [Focusing example – bright field] - Focusing mode ③ Focus option * Enlarged image Bright- Good Focus Bright- Bad Focus #1 Bright- Bad Focus #2 Bright- Bad Focus #3 • Good Focus: Compared to gray background, dark inside of cells and bright boundaries around cells.
  • Page 31 CD45 (PerCP)- Bad Focus CD34 (PE)- Good Focus CD34 (PE)- Bad Focus • Good Focus: Clear cell image, bright cell body in contrast to the dark background. • Bad focus: Unclear cell image, blurry cell image. ADAMII™ © NanoEntek, Inc.
  • Page 32 Measurement Measure menu 1. Before running samples, make sure that sample type and dilution factor are selected accurately. - Focusing mode ④ Run sample 2. Press the “Run sample” button. 3. Enter the Sample ID, Patient ID, Antibody Lot, and Slide Lot. 4.
  • Page 33 There is no need to use this button after the assay is completed ordinarily. The completed Assay Slide will be automatically ejected at the end of the measurement. ☞ Note: The slide door should be closed at all times except when inserting and removing the slides. ADAMII™ © NanoEntek, Inc.

  • Page 34: Data

    Data Data menu Data menu provides the counting results of a measurement, the countings of each channel at each frame, the lists of data, and an option to export data. 1. Test information 2. Sample information 3. Captured image 4. Sample volume information 5.
  • Page 35 • Measured frame: Number of frames that were measured. • Deleted frame: Number of frames that were deleted. • Resulted frame: Final number of frames that were used for the results. (The number of measured frames subtracted by the number of deleted frames) ADAMII™ © NanoEntek, Inc.
  • Page 36 Data Data menu In this window, raw image in each channel at each frame can be ③ Captured reviewed. Click left or right arrow, or enter a number to choose a frame to see images. Click a channel button to see the image in image that channel.
  • Page 37 Double click the captured image, then you can check preview images ③ Captured of selected channel on the right side of the screen. To check the pre- view images of other channels, select that channel and double click image the captured image again. ADAMII™ © NanoEntek, Inc.
  • Page 38 Default values are listed below. Please refer to the package insert included in ADAMII-CD34 kit. • HPC-A: Total volume = 275 μL, sample volume = 20 μL • Mobilized PB: Total volume = 60 μL, sample volume = 20 μL •...
  • Page 39 Frame by frame revision of raw data image and removal of invalid frames can be done on the table. For deleting an invalid frame, click on the right mouse button and select ‘Delete’ menu. Then, the “Result area” will show the re-calculated data, reflecting the deleted frame(s). ADAMII™ © NanoEntek, Inc.
  • Page 40 Data Data menu How to find the invalid frames is described below: ⑦ Raw data table 1) Sample aggregation: When there is aggregation in a specific area of the sample, that frame should be deleted. Br image FL image 2) Dust: When there is dust inside the slide, that frame should be deleted.
  • Page 41 4) Cell debris: When there is cell debris, that frame should be deleted. ⑦ Raw data table Br image FL image 5) Overlapped circle: When there are several circles in one cell, that frame should be deleted. Br image FL image ADAMII™ © NanoEntek, Inc.
  • Page 42 Data Data menu In the Data list, all previous measurements are shown in a chronological ⑧ Data list order. Scroll up and down to find the data of interest and select the data to review. ⑨ Edit data list User can edit the Data list. •...
  • Page 43 • csv file: Data table containing comprehensive information in a spreadsheet. • jpg file: Raw images of all 4 channels. • pdf file: A report with summaries of measurements including CD34+ cell counts, CD34+/CD45+ ratio, and viability. [csv file] [jpg file] [pdf file] ADAMII™ © NanoEntek, Inc.

  • Page 44: Setting

    Setting Setting menu Setting menu contains information of ADAMII-CD34 software as well as several user preferences and other Settings. It allows users to change settings including cell counting settings, and update software and firmware. 1. Date & Time 2. Counting settings 3.
  • Page 45 Setting Setting menu You can change date and time of ADAMII-CD34 software. ① Date & Time ② Counting It is not recommended to adjust the sensitivity and circularity by the user. settings • Sensitivity : Refers to the contrast of the objects from the background.
  • Page 46 Setting Setting menu Please refer to page 12~19 for calibration. ③ Calibration To open the lists of calibration histories, click “Record” button. User can see the records of previous calibrations. ④ Sample While we don’t recommend users to change this, user can change the default sample volume and total volume.
  • Page 47 [Software update] 1. We recommend to contact NanoEntek customer service (ivdst@nanoentek.com) if SW or FW needs to be updated. 2. If an update version of SW or FW has been provided, copy the file in a USB drive and plug it into the laptop.
  • Page 48 Setting Setting menu Device info indicates serial number of the ADAMII-CD34 and the status of ⑦ Device info connection between camera in the instrument and the laptop. ⑧ Disk info Disk info indicates available space in the hard drive installed in the laptop.

  • Page 49: Cleaning & Maintenance

    Cleaning and Maintenance Clean the surface of ADAMII-CD34 with a damp cloth. If liquid is spilled on ADAMII-CD34, turn off the power immediately and wipe out liquid. ADAMII-CD34 does not need regular maintenance. To troubleshoot problems with ADAMII-CD34, contact technical support.

  • Page 50: Troubleshooting

    Troubleshooting Installation • Check on/off switch on back side of main instrument. ADAMII™ • Check power source or contact your distributor. does not power on Operator program • Check on/off switch on back side of main instrument. does not start •...
  • Page 51 • Do not use any other slides as it results in inaccurate count and may damage the instrument. Poor image quality • While viewing cells in preview state, re-optimize the focus. ADAMII™ © NanoEntek, Inc.

  • Page 52: Error Code

    Error code Error code Reason Message Position Camera E0001 connection fail Driver Serial E0002 connection fail Serial Stage E0010 disconnected working Camera E0020 Camera disconnected. Slide holder is Slide E0030 opened. inserting Slide door is Slide E0031 Slide door is opened. opened.
  • Page 53 Error code Error code Reason Message Position Data storage E0040 Data save is full Sample density Measuring E0050 is too high E0060 Update fail SW update ADAMII™ © NanoEntek, Inc.

  • Page 54: Warranty

    Warranty NanoEntek provides 1-year warranty service for defects of material and workmanship. If any defects occur in ADAMII™, NanoEntek provides repair services for the defective parts at its discretion. The following defects, however, are specifically excluded: 1. Defects caused by improper operation.

  • Page 55: Safety Precautions

    In the documentation for the user, a statement shall be included drawing attention to the fact that there may be potential difficulties in ensuring electromagnetic compatibility in other environments, due to conducted as well as radiated disturbances. ADAMII™ © NanoEntek, Inc.

  • Page 56: Consignes De Securite

    Consignes de securite Examinez et suivez les consignes de securite ci-dessous : · N’installez pas l’instrument dans un endroit humide comme une serre ou un incubateur pour eviter un risque de choc electrique. Si de l’eau ou tout autre materiau penetre dans l’instrument, l’adaptateur, ou l’entrees d’alimentation, debranchez le cordon d’alimentation et contactez un tech nicien de service.

  • Page 57: Safety Symbols

    “Safety Requirements for Electrical Instrument for Measurement, Control, and Laboratory Use, Part l: General Requirements.” This instrument bearing the TÜV symbol are certified by TÜV Product Services to be in conformance with the applicable safety standard for the US and Canada. ADAMII™ © NanoEntek, Inc.

  • Page 58: Product Specifications

    Product specifications ADAMII-CD34 Bright, Green (PE), Green (PerCP) and Blue (FITC) Light source Camera High-sensitivity monochrome CCD Automated X-Y-Z stage Stage CSV, PDF, JPEG Exported formats: Image 12VDC, 5.0A Electronic input Operating power 100 – 240 V, 1.5 A, 50/60 Hz 5 - 40 °C, 20 –...

  • Page 59: Ordering Information

    · Main instrument ADAMII , Bench-top · Power cord ADAM2 Fluorescence Cell Counter · USB connection cable · ADAMII-CD34 Reagent · 10X RBC Lysis Buffer ADAMII-CD34 Kit CD34K-025 · ADAMII Calibration Beads · ADAMII Assay Slide ADAMII™ © NanoEntek, Inc.

  • Page 60: Technical Support

    Technical support Visit the our Website at www.nanoentek.com for : · Technical resources, including manuals, FAQs, etc. · Technical support contact information · Additional product information and special offers For more information or technical assistance, please call or email. Email ivdst@nanoentek.com...
  • Page 61 ADAMII™ © NanoEntek, Inc.
  • Page 62 851-14, Seohae-ro, Paltan-myeon, Hwaseong-si, Gyeonggi-do, 18531, Korea Tel :+82-2-6220-7942 , Fax:+82-2-6220-7999 MT Promedt Consulting GmbH Altenhofstrasse 80, 66386 St. Ingbert, Germany NanoEntek America, Inc. 220 Bear Hill Road, Suite 102, Waltham, MA 02451, USA Tel: +1-781-472-2558 , Fax: +1-781-790-5649 Email : ivdst@nanoentek.com...

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