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NanoEnTek JuLI Stage User Manual

Real-time cell history recorder
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NanoEnTek Inc. All rights reserved.
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JuLI™Stage, Real-Time Cell History Recorder

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  • Page 1 User Manual Copyright NanoEnTek Inc. All rights reserved. JuLI™Stage, Real-Time Cell History Recorder...
  • Page 2 The information in this manual is described as accurately as possible. The changes involved in firmware and software may be applied without notification. Copyright ©2014 by NanoEnTek Inc. All rights reserved. Published in Korea. Documentation: NESMU-JST-001E Revision history: V.0.0 Date: OCT 2014 V.0.5 Date: MAY 2015...

  • Page 3: Table Of Contents

    Table of contents Product components Quick operation guide Product overview - Position Calibration (for Vessel type) Features - Stitching Image Calibration (Stitching & Map mode) Description - Image capture, save and overlay - Operation type-1 - Front view (for Selecting multiple samples and use the Auto focus) - Side view - Operation type-2 - Rear view...

  • Page 4: Product Components

    The JuLI™ Stage, Real-Time Cell History Recorder is shipped with the following components. Please check all components listed below when the instrument is delivered. PC will be shipped separately because it is optional. If any item is missed of damaged, contact your local distributor or e-mail sales@nanoentek.com. JuLI Stage main instrument...

  • Page 5: Product Overview

    Product overview A fully-automated, real-time cell history recorder that acquires cell images directly from various cell culture plates (6 to 384 well) slides and dishes in an incubator. JuLI™ Stage supports multi-channel fluorescent colors, multiple objective lenses, and sensitive filter-based optics to optimize for various live cell assays. In addition, it enables users to support quantified cellconfluence results with low variation and the growth curve using image-based analysis with bright field.

  • Page 6: Features

    Features The JuLI Stage a Real-Time Cell History Recorder, offers you the following benefits, Automated imaging system The JuLI™ Stage provides a live cell imaging using automatic focus and a precise one by one multi-well control system. - Fully automated and motorized X-Y scanning stage - Auto focusing on every single position - Image tiling with a stitching function Sensitivity and expandability...

  • Page 7: Description

    Description Cover Used to protect test samples from external light. When you use the JuLl™ Stage Front view in an incubator, the cover can block out light from the outside. Main body Front view of the JuLl™Stage. The main body is composed of LED, Filters (Fluorescence mode) and optic parts.

  • Page 8: Rear View

    Description Rear view Machinery cooling system. Connection port Connect the JuLI™ Stage main instrument with the control box. Power inlet Connect the instrument to an electrical outlet using the supplied power cord and the appropriate plug, based on the electrical outlet configuration in your country.

  • Page 9: Installation

    Installation 1. Open the box and remove the material foam from the box. Unpacking 2. Carefully lift the instrument out of the box, holding it by handholds in the bottom of the base. the instrument 3. Place the instrument on a flat, level surface (table or incubator). IMPORTANT! Do not lift the JuLI™...

  • Page 10: Install The Instrument

    Installation 1. Connect repeater to the PC and two USB cable, then connect to control box. Install (PC Monitor Resolution: 1920 ×1080 pixels) the instrument ☞ Note: Check the connection on the rear side of the PC. ☞ Note: If you connect all the components, it will look like the Image below. 1-1.
  • Page 11 Installation 4. Turn the PC and the Control box on by pressing the Power button. Install the instrument 5. Double-click the JuLI™ Stage icon on Desktop. The JuLI™ Stage software starts up. ☞ Note: The JuLI™ Stage will be ready to use after initialization is completed when power on. JuLI™Stage, Real-Time Cell History Recorder...

  • Page 12: Place The Instrument

    Installation Live-cell images from various cell culture dishes are directly captured in a cell culture incubator. Place the This design allows an instrument to be installed in an incubator, and prevents the contamination instrument into by maintaining a sterile environment. The intuitive graphic user interface also provides quick and the incubator convenient use in a cell culture incubator while capturing time-lapse imaging.

  • Page 13: Operating Environment

    Installation • Allow at least 10 cm (4 inches) free space at the back of the instrument to allow for proper venti- Operating lation and prevent overheating of electronic components. environment • Set up the JuLI™ Stage away from direct light sources, such as windows. Ambient room lighting can enter the imaging path and affect the image quality.

  • Page 14: User Interface

    User interface The JuLI™ Stage software allows you to generate live cell imaging and analysis easily. - Image capturing and time-lapse recording with multi-position scanning - Navigation function to figure out the optimal position - Image stitching function to scan the whole well or regions of the well at once The JuLI™...
  • Page 15 User interface Data menu Settings menu JuLI™Stage, Real-Time Cell History Recorder...

  • Page 16: Basic Operation

    Basic operation The JuLI™ Stage can be used in combination with PC (Monitor Resolution 1920x1080) enabling users to operate the instrument using the computer mouse and keyboard. The JuLI™ Stage basic operations use the mouse for moving, selecting and zooming in/out. The detailed explanations are listed below.

  • Page 17: Selection

    Basic operation 1) Selecting well Selection The well is marked by a grey color when the specific well is selected by clicking The Left button. Click once to select and click once more to deselect. ☞Note: The JuLI™ Stage supports selecting function only in the Option menu. The selecting function does not work in the Preview menu.
  • Page 18 Basic operation 2) Selecting position Selection When the specific position well is selected by clicking the Left button, the assigned position is marked by a Green square in the Navigation window. Press the Position set button in order to confirm. The JuLI™...

  • Page 19: Position Zoom

    Basic operation Double click the position to delete. Selection Press the Position Add button to save your setting - the green square will change to a Red square. Left mouse 1X zoom in Position zoom 2X zoom in 4X zoom in 8X zoom in When you press + or - button, it activates the zoom function.

  • Page 20: Zoom

    Basic operation 1) Zoom-in Zoom Scroll up in the view field to use the zoom in the image. The JuLI™ Stage supports zoom-in function in both the real view and captured images allowing users to magnify specific position by scrolling up. Mouse wheel (Up and down) <...

  • Page 21: Focus

    Basic operation 3) Focus adjustment Focus Scroll up or down with Shift key on the keyboard for fine focus adjustment. It is the same function with the Fine focus interface in the menu Coarse focus interface Mouse wheel Fine focus interface (Up and down) ☞...

  • Page 22: Auto Focus

    Basic operation Auto Focus Toggle button is used during the monitoring. Auto Focus It activates auto focus between wells. Select the number of wells to skip from the current well. Set the number based on well. According to the number, auto focus will be activated on wells which the user wanted, this can be adjusted allowing for flatness of plate/sample.
  • Page 23 Basic operation Auto Focus Auto Focus When you set 5 in Auto Focus, activate auto focus on wells with skipping 5 wells. ⇒ So, only the while colored wells will activate auto focus. All Position Toggle button activates auto focus between positions. Select the number of All Position how many positions to skip from the current position.
  • Page 24 Basic operation Auto Focus All Position When you set 3 in All Position, activate auto focus on positions with skipping 3 positions. ⇒ So, only the red square positions will activate auto focus. When you set 5 in All Position, activate auto focus on positions with skipping 5 positions. ⇒...
  • Page 25 Basic operation Tip: When you set the positions leaving spaces, right next to the positions are an adjacent Auto Focus focus area. Therefore, activate auto focus only at the one position and save time. All Position There are 6 positions. If you set 2 in All Position, only the 3 red square positions will acti- vate auto focus.
  • Page 26 Basic operation All Channel Toggle button activates auto focus in all channels. Auto Focus All Channel Activates auto focus on Bright field channel. When you turn on All Channel, auto focus will also activate the GFP/RFP/DAPI channel as per Auto Focus and All Position setting that user specified.
  • Page 27 Basic operation Steps are as below. Auto Focus Custom AF 1. Adjust the focus according to your sample, and then press the Custom AF button (When the autofocus mode is turned on). 2. The JuLI™ Stage Software automatically adjusts the Optimal focus corresponding to your sample in the bright field.

  • Page 28: Operation

    Operation The Preview menu allows you to select basic system functions and is the first screen after Preview menu start-up. It is available for checking the sample, setting the optimized focusing, capturing and merging the images. Vessel type button Position Zoom in & out Position List button Map mode button Navigation window...
  • Page 29 Operation 1. Press the Vessel button. Preview menu 2. Select the Vessel type, which is used for selecting the sample vessel category. Vessel type 3. Press the Vessel button again to close the window without calibration. 1. Press the Calibration button, if necessary. Vessel 2.
  • Page 30 Operation 1. Press the Map mode button. Preview menu 2. Press the Make Map button which will appear after pressing the Map mode button. Map mode 3. Select the LED channel, map range which is free to choose (1 X 1 ~ 99 X 99), and check whether or not to use auto focus function.
  • Page 31 Operation 1. Preview well is marked by a Yellow circle. Preview menu 2. The field you selected is marked by the Red square in the Navigation step. Navigation window ☞ Note: The size of red square, which is field of view, depends on vessel type. 24 well plate 96 well plate ☞...
  • Page 32 Operation Camera setting allows users to customize the user interface within the range of Preview menu setting values. To adjust the illumination intensity, use the Exposure and Brightness Camera set bar. To adjust the power of light sources, use the LED power bar. To observe in stereoscopic, use the D-Phase bar.
  • Page 33 Operation 1. Press the Focus information button. Preview menu 2. The focus information button is designed to provide the focus setting information Focus about each well. information e.g. A1: Complete Bright focus A2: Complete Bright and GFP focus A3: Complete Bright, GFP and RFP focus Press the Scale Bar button to mark the Scale Bar on captured image.
  • Page 34 Operation Press the Save button to save the captured image. Preview menu Enter the file name and store the path when the pop-up window appears. Save image Real time preview recording function ( 10 FPS, maximum 55min) Record It will be recording about the current viewing situation.
  • Page 35 Operation 1. Capture images from more than two channels. (Bright, GFP, RFP or DAPI). Preview menu 2. Press the Overlay button, then the editing window will appear. Overlay image 3. Press the Save button in the overlay window to save the overlapped image. Press the Stage moving control button to control the X-Y stage position.

  • Page 36: Option Menu

    Operation The Option menu is composed of necessary functions in cell monitoring. It is divided into two Option menu parts (camera setting option and monitoring setting option). setting option ☞ Note: Please refer to the details of camera setting option in preview menu. Vessel type button Navigation window Position Zoom in &...
  • Page 37 Operation Select All : Press Select All button to select all wells. Option menu Navigation Position Add : Once Position Add button is pressed after assigning an area you position set want, the area will be changed into Red square at navigation window. Default button : Select well and then press Default button, center value of well will be assigned.
  • Page 38 Operation Option menu Project Title To start monitoring, assign the title using the pop-up keyboard. Press Protocol button to import the saved X, Y coordinate value. Protocol Vessel Type : Select Vessel type. Time Range : Select All Data to find data from the whole days. Select Days ago (Last Used) to classify recent data.
  • Page 39 Operation Manual Focus Toggle button is used during the monitoring. Option menu If there is Z value of a set position, move the Z coordinate and locate. Manual Focus All of the functions listed below allow users to do specific auto focus on well and position as Auto Focus required.
  • Page 40 Operation If you seletct depth and number of stack images, you can obtain images of various Option menu focus from one position (above and below set focus plane). Z stack Number of images is from 3 to 21, and depth range is from 0.5µm ~ 10µm Image format can be saved as 4 types of file.
  • Page 41 Operation Press the Start button to move to the next step. Option menu Start button Press the Cancel button to reset. Cancel button IMPORTANT! Warm up the JuLI™ Stage for 2 hours before monitoring. The power should be on when warming up the instrument. The well plate’s cover should be wetted by the cell culture media before monitoring.
  • Page 42 Operation While the instrument operates, you can control the monitoring cycle number. Option menu Detail methods are as below. Edit the cycle number 1. Click cycle number display box during the instrument operating. 2. When keypad appears, enter the cycle number that you want to edit. If you press OK button, the value will be applied to cycle number display box.
  • Page 43 Operation While capturing Timelapse current filmed condition, completed image Option menu and others are expressed in Process bar. Process bar Image and graph which captured at selecting past area are also displayed. While the instrument operates, if you use All well button, you can ob- Option menu serve images of whole well at the same time.

  • Page 44: Data Menu

    Operation Data menu provides the function to view and export the captured images. In the data menu us- Data menu ers are able to check. Once you selected the title, the JuLI™ Stage shows test conditions including vessel type, well captured position, LED channel condition, interval time and cycle number.
  • Page 45 Operation The selected title is marked by green color when clicking a title. Data menu Title list The color of vessel contains information about test conditions. Vessel display Grey circles: Captured well Red circles: Captured well and selected for review LED set &...
  • Page 46 Operation Time setting condition and the mode information are displayed. Data menu Time set & Mode Press the Export button to export the captured images. The multiselection function is supported. Display export ☞ Note:Access or download the image data through JuLI ™ Stage / TIMELAPS folder in PC.

  • Page 47: Setting Menu

    Operation Setting menu contains information about the JuLI™ Stage as well as several user preference and Setting menu other Settings. Setting menu allows users to edit settings about the software & firmware informa- tion and basic operations. Serial number bar Lens information Data path information Mouse control buttons...
  • Page 48 Operation Shows serial number of the JuLI™ Stage main instrument. Setting menu Serial info Provides lens information when it moves and checks the size. The lens can be changed manually Objective Lens and the size of field of view (square) on navigation window is also changeable with lens informa- tion.
  • Page 49 Operation The JuLI™ Stage Cell History Recoder activation. Setting menu Activation Administration mode. It is for manufacturing engineers to set the system. Admin mode The JuLI™ Stage checks for the latest version of software or firmware and automatically system info displays the information window whenever it starts up.

  • Page 50: Quick Operation Guide

    Quick operation guide Table of contents 1. Position Calibration (for Vessel type) 2. Stitching Image Calibration (Stitching & Map mode) 3. Image capture, save and overlay 4. Operation type-1 (for Large scale of sample using the Auto focus) 5. Operation type-2 (for small scale of sample using the Manual focus)
  • Page 51 Caution IMPORTANT! Cautions The JuLI™ Stage is optimized for a standard size of cell culture incubator. It is recommend that the temperature should be stabilized in case of a small size incubator before using the instrument. Warm up the JuLI™ Stage for 2 hours before monitoring. The power should be on when warming up the instrument.

  • Page 52: Position Calibration (For Vessel Type)

    Position Calibration (Vessel type) 1. Press the Move To A01 or B02 button. Like the picture, stage will move to A1 or B02 well and the center square displays the current location. 2. Press the Check A01 or B02 button. If you press the button, The Stage will move to 4 quad- rants (top, bottom, left and right) to check positioning around the radius of the well.

  • Page 53: Stitching Image Calibration (Stitching & Map Mode)

    Stitching Image Calibration (Stitching & Map mode) If there is overlapped image during map mode or stitching, need this function. 1. Select Make Map of Map Mode from Preview tap. 2. Press Calibration button in the Make Map. 3. Select Horizontal Axis Mode button. 4.

  • Page 54: Image Capture, Save And Overlay

    Image capture, save and overlay 1. Select the Preview menu. 2. Press the Vessel button. 3. Select the Vessel type according to your sample when well type list appears. 4. Press the Calibration button, if necessary. Set the Calibration option from first well to end well. 5.
  • Page 55 8. Find optional focus and adjust with Focus bar and buttons 9. Press the Capture button to start capturing images. 10. Press the Save button to save the captured images, if necessary. 11. Once pop-up window appears, assign the storing path and file type, and press the Save button to confirm.

  • Page 56: Operation Type-1

    Operation type-1 (for selecting multiple samples and using the Auto focus) 1. Select the Option menu. 2. Press the Vessel button. 3. Select the Vessel type according to your sample when well type list appears. 4. Check the Selected Vessel type and press Vessel button again to finish setting.
  • Page 57 9. Press the Default button to equally capture central position of all wells. 10. The assigned position is marked by green square on navigation window. 11. Press the Position Add button to complete setting - the green square is changed to Red. 12.

  • Page 58: Operation Type-2

    Operation type-2 (for selecting Individual samples using the Manual focus) 1. Click a well you want to capture using right mouse button. 2. Once the well is selected, the X-Y stage is automatically moved to center of well. Use Right mouse button to navigate around the well to discover points of interest within the well.
  • Page 59 8. Select Other LED channels, If necessary to capture additional images in different channel. Follow Step 4 ~ 7 repeatedly and save images for each channel. 9. After adjusting focus and assigning position, press the Position Add button. 10. Press the OK button when the pop-up window appears.

  • Page 60: Cleaning And Maintenance

    Unauthorized repairs may damage the JuLI™ Stage or alter its functionality, which will void your warranty. Contact sales@nanoentek.com or your local the JuLI™ Stage distributor to arrange for service. The JuLI™ Stage is optimized for a standard size of cell culture incubator.

  • Page 61: Troubleshooting

    Troubleshooting Intstallation JuLI Stage does not Check on/off switch on back side of main device. power up Check power source or contact your distributor. LCD screen is black Move the mouse. Check if power supply is connected and power switch is on.

  • Page 62: Warranty

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

  • Page 63: Safety Precautions

    Safety precautions Review and follow the safety instructions below : If water or other material enters the instrument, adaptor, or power inlet, disconnect the power cord and contact a service person. For operating environment, refer to Product Specifications. Do not touch the main plug or power cord with wet hands. Always ensure that the power supply input voltage matches the voltage available at your location.
  • Page 64 Safety precautions Review and follow the safety instructions below : Symbol Meaning Caution and Warning Protective earth (Ground) This instrument and consumables conforms to the EC Declaration of Conformity. This equipment has been tested and found to comply with the limits for a Class A digital instrument, pursuant to Part 15 of the FCC Rules.

  • Page 65: Product Specifications

    Product specifications Items Specification Light source Blue, Green, UV LED (Intensity adjustable) Objective lens 4 X, 10 X & 20 X + Digital Zoom Inter-changeable objective lens Fluorescence 3 fluorescences DAPI: Excitation 390/40, Emission 452/45 GFP: Excitation 466/40, Emission 525/50 RFP: Excitation 543/22, Emission 580LP Camera High-sensitivity monochrome CCD (Sony sensor 2/3”)

  • Page 66: Ordering Information

    JuLI™ Stage, Vessel holder T-Flask 1 pc, Holds one T-25 flask 1 pc, Holds one T-75 flask (25 & 75 cm ), (Optional) JOS1000 JuLI Stage Software (CHR) CHR (Cell History Recorder) SOS1000 JuLI Stage Software (SOS) SOS (Single-Cell Observation System) JuLI...

  • Page 67: Technical Support

    For more information or technical assistance, please call or email. NanoEnTek Inc. 851-14, Seohae-ro, Paltan-myeon, Hwaseong-si, Gyeonggi-do, 18531, Korea Tel : +82-2-6220-7940 Fax : +82-2-6220-7999 NanoEnTek Inc. (USA) 240 Bear Hill Road, Suite 101, Waltham, MA 02451, USA Tel: +1-781-472-2558 Fax: +1-781-790-5649 Email sales@nanoentek.com...
  • Page 68 NESMU-JST-001E (V.2.0) NanoEnTek Inc. 851-14, Seohae-ro, Paltan-myeon, Hwaseong-si, Gyeonggi-do, 18531, Korea Tel : +82-2-6220-7940 Fax : +82-2-6220-7999 NanoEnTek Inc. (USA) 240 Bear Hill Road, Suite 101, Waltham, MA 02451, USA Tel: +1-781-472-2558 Fax: +1-781-790-5649 Email sales@nanoentek.com Website www.julistage.com www.nanoentek.com...

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