CURIOSIS Celloger Mini Plus Instruction Manual Download Page 1

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Celloger® Mini Plus

Automated live cell imaging system

Instruction Manual

Summary of Contents for Celloger Mini Plus

Page 1: ...Celloger Mini Plus Automated live cell imaging system Instruction Manual ...

Page 2: ...SIS Inc 4F 10 Teheran ro 38 gil Gangnam gu Seoul 06221 South Korea Tel 82 2 508 5237 Fax 82 2 508 5246 Copyright 2022 by CURIOSIS Inc All rights reserved Published in Korea CR IFU 008 Rev 2 FOR RESEARCH USE ONLY and not for use in diagnostic procedures Specifications subject to change without notice ...

Page 3: ...apse imaging in Scan App 24 5 Process single image or time lapse images in Analysis App 25 5 1 Import the original data taken by Celloger Mini Plus 25 5 2 Merge FL image 26 5 3 Slide show 26 5 4 Make video 27 5 5 Estimate confluency coverage 27 5 6 Create confluency graph 29 5 7 Show Z stacking image 29 5 8 Stitch images 31 5 9 Separate the image 34 V Software description 35 1 Celloger Mini Plus S...

Page 4: ...for dish 35mm dual Optional 1 Vessel holder for dish 60mm dual Optional 1 Vessel holder for dish 90 100mm single Optional 1 Vessel holder for T flask 25cm2 single Optional 1 Vessel holder for T flask 25cm2 dual Optional 1 Vessel holder for T flask 75cm2 single Optional 1 When receiving the package Check that all items listed above are included in your package Examine the device carefully for any d...

Page 5: ...5 II Device layout ...

Page 6: ...of the Celloger Mini Plus If using multiple equipment there should be a proper clearance among equipment Keep the device at temperature between 10 40 C Relative humidity range is 20 95 Important It is not recommended to operate the device at low temperature Below 10 C In those conditions warm up the device for over 10 minutes PC specifications p 55 Celloger Mini Plus software can be used on PCs wi...

Page 7: ...r the incubator 2 Connect two LAN cables and power cable to the POE injector Connect one LAN cable to the device yellow and another LAN cable to the PC white 3 Connect power cable to an outlet 4 Turn on the power by pressing the power switch located next to the power cable port and check if the light indicator turns on Install the device inside an incubator 1 Install the vessel holder on Celloger ...

Page 8: ...ues Light intensity Focus position Schedule File storage location etc are saved in the project Note In Celloger Mini Plus Analysis App the setting is managed by config file instead of project file Create a new project or open an existing one You can start a program with default project by clicking Continue without configuration Import the project saved in the past Create new project Start the prog...

Page 9: ... address 5 Connect the device 1 Open Celloger Mini Plus Scan App 2 When the app is executed Create or Open a project popup appears Select Continue without configuration 3 Connect your device popup will show 4 Click Get Network Adapter to load the network adapters connected to the PC 5 Select PC network from Network adapter list then the device icon will appear on the right 6 Click the device icon ...

Page 10: ...rectly from Case 4 When using multiple devices in one PC each device has the same IP Initially devices are released with the same IP When using the device for the first time it is necessary to reset the IP address refer to p 57 for device IP setting 7 Click Connect When the connection has successfully completed the LED status indicator lights in green from yellow Device connection pop up descripti...

Page 11: ... and time lapse imaging function Celloger Mini Plus Analysis App is a program for analyzing and post processing the images obtained from Caloyer Mini Plus and it can be operated even without the device connection Note Refer to p 35 for full software descriptions 6 1 Celloger Mini Plus Scan App Menu bar Toolbox Display Preview Light source control Device status Jog control Vessel Plate panel Scan p...

Page 12: ...12 6 2 Celloger Mini Plus Analysis App Menu bar Toolbox Display Light source control Z coordinates Image analysis control ...

Page 13: ...e cell and Trypsin EDTA to conical tube and add fresh medium to neutralize the Trypsin reagent 7 Centrifuge the suspension cells on conical tube 8 Discard the supernatant and add fresh medium into cells 9 Gently pipette the cell several times 10 Transfer the appropriate amount of cells into new vessel and dilute the cell concentration by fresh medium 11 Cover the plate with a lid and gently shake ...

Page 14: ...te Celloger Mini Plus Z stage absolute coordinate error is 0 001 mm Upon finding the best focal point for scanning designate the location by pressing Current Position in Set Position Note If you want to change the focus select the position you wish to change in the scan table click under Properties and click on Apply or Apply All to apply the change refer to p 45 for Properties Note When Auto Focu...

Page 15: ...point and Z position coordinates from autofocusing IsDarkSideOriented Select whether to focus on the black side when performing autofocus Focus is on black side when checked if not focus is on the clearer side of either black or white Note The larger Depth and the smaller Resolution the higher autofocusing accuracy Meanwhile the longer the screening time the longer autofocusing time Set the value ...

Page 16: ...reen once selected Select the FL channel in Light Source Control and adjust the focus using the Jog button by referring to the Display Check that the FL Focus Offset checkbox under Properties is checked and click on to specify the focus and click Apply All If the position is specified after checking in the FL mode the FL Focus Offset checkbox is checked Check if the designated FL Focus Offset valu...

Page 17: ...Z stacking 1 Manual Z stacking By using Jog find the position to start stacking Designate the stacking unit in the Step Ex Step setting for stacking by 10 𝜇m In the Jog button click Current Position for each position of the range you want to stack by using 2 Automated Z stacking Select the position in the Scan table where the z stacking function will be applied Select whether or not to execute the...

Page 18: ...e the stitching function will be applied Select whether or not to execute the stitching function in BF and FL in Stitching under Properties refer to p 46 for Stitching Set the overlapping area between the images using Overlap scroll bar and designate the range to be scanned in Range Properties Z Stacking Scan table Properties Stitching ...

Page 19: ...FL Lamp 30 Exposure 0 5 Gain 1 3 Image analysis in Scan App 3 1 Capture icon in Toolbox captures the image on Display Use Ctrl I as a shortcut 1 An image with features It is possible to generate and manipulate the desired features show it on Display and save it as it is To obtain without features press Features layer for deactivation and then press 2 BF or FL channel image Press the desired tab of...

Page 20: ... thresholding Divide the image into local areas and thresholding is conducted by comparing relative intensity among areas Adjust parameters of the Kernel Size and Offset while looking at the mask of the image after checking the Adaptive Algorithm box Kernel Size indicates size of local and Offset indicates the relative offset of the threshold of local area Note When the fluorescence areas are dist...

Page 21: ... menu and save the modified image Note In case of fluorescence as shown in the image below Bad Pixel Correction and Brightness Contrast are set by default therefore it should be adjusted Note The adjustments are made with captured images so cells are not affected during scanning Before adjustment After adjustment ...

Page 22: ... Record 3 6 Intensity 1 Point Intensity Mean Intensity Upon activating Intensity in Visibility menu Point Intensity and Mean Intensity information is displayed at the upper left of Display Intensity at X coordinate Y coordinate Point intensity value of the X Y coordinates indicated by the cursor If it is in merge state intensity of BF channel and intensity of FL channel are displayed in order Mean...

Page 23: ...he items to be deleted To delete all press Clear To hide them without removing all data deactivate them by pressing Manual Counting activated in Visibility of Tool menu 3 8 Ruler In Measure select either Free Horizontal or Vertical directions and select the desired area from point to point The number on the line is the length measurement 𝜇m Note To undo the selected point after selecting one point...

Page 24: ...rescence is also scanned the focus of fluorescence should also be adjusted Open Time lapse Control and select the location to save and enter the project name If the image storage location is not specified the file will be saved in the Scan folder under the Celloger Mini Plus Scan App folder and if the project name is not entered a folder will be automatically created under the scan start date Set ...

Page 25: ...re the merged image and raw images of reference position followed by Z stacked images of BF and FL are shown alternately from the low to high Z axis Stitching When image is captured with only BF channel or BF and FL channels the first 2 or 3 images are merged image and raw images same as Z stacking followed by the images in scanning order Refer to p 47 for the stitching scanning order 5 Process si...

Page 26: ...ge file Note Folder can be dragged Note The file location address and file name should only be composed of English alphabets number and some special symbols a file name shouldn t contain any of the following characters to import the file correctly in Celloger Mini Plus Analysis App If it fails to import the file check whether the file name and location address is written in languages other than En...

Page 27: ...irmation Note In order to change the speed change the parameter in Preferences of Tool menu Recommended value 5 13 Note FPS is a parameter indicating the speed of slide show and video However the speed of slide show and video could be different even with the same FPS 5 5 Estimate confluency coverage Open the file and images captured with Celloger Mini Plus Press Analyze To analyze certain images f...

Page 28: ...ndicated When a cycle is selected from the list the corresponding cycle s image and confluency mask can be seen Note If you wish to adjust the confluency masking color or transparency select Preferences under the Tool menu change the parameters in Analysis under Merge Layer and click Apply Note If you do not want to see the confluency mask deactivate it by clicking on under Toolbox Calculated conf...

Page 29: ...tions are displayed at the same time The image below shows the confluency graph when selecting 3 positions among 12 scan positions 5 7 Show Z stacking image Open the time lapse file or images captured with Z stacking function After selecting the scan position with Z stacked images click the Z stacking tab under Image Analysis Control to see the Z stacking list The image below shows the Z stacking ...

Page 30: ...ay ZStacking 2 means the image was taken two steps below the reference position Focus For instance if the Step is set to 10 𝜇m ZStacking 2 is taken 20 𝜇m below the reference position refer to p 17 for more information of Z stacking setup Note If you want to make a video from the images of the Z stacking list click Record on the Z stacking tab The video appears in the order of the Z stacking list f...

Page 31: ...ges click the Stitching tab under Image Analysis Control to see the Stitching list The image below shows the Stitching list when the Range of stitching function is set to 1 1 1 1 Reference Position Overlap Overlap Stitching 1_ 1 Stitching 1_0 Stitching 1_1 Stitching 0_1 Stitching 0_ 1 Stitching 1_ 1 Stitching 1_1 Stitching 1_0 Stitching list ...

Page 32: ...the fourth cycle captured at the first scan position on Display Stitching1_ 1 means the image was taken one step right and one step down from the reference position overlapped by the value set in overlap Note Thread refers to the number of images taken to perform the stitching function horizontal x vertical Stitching Channel shows the light channel selected when stitching was performed Scan positi...

Page 33: ...ted and Success is displayed in the Result When ViewChange is clicked stitched image appears on the Display Note When stitching fails Fail appears in the Result Click Save to save the stitched image Note Be reminded that the image shown on the Display will be saved so be sure to check the display image before clicking Save ...

Page 34: ...d and saved as individual image files When Separate is selected from Folder File tab the raw images BF FL of the reference position is separated and saved When clicked from the Z Stacking or Stitching tab images from each list will be separated and saved For example if you click Separate on the Stitching tab the images in the Stitching list are made into individual image files and saved refer to t...

Page 35: ...t Ctrl S can be used Save Project As Create a new project with current settings Save Image As Save the image shown on the Display Keyboard shortcut Ctrl I can be used Histogram p 23 Show hide intensity histogram of an image Intensity p 22 Show hide point and mean intensity of an image Manual Counting Show hide Manual Counting function Scale Bar Show hide scale bar on the left bottom of Display Cen...

Page 36: ...nnected device Parameter Display the values set for the connected device Certain parameters can only be manipulated by an authorized person IPAddress Change the IP address of the connected device F W Update Update the firmware of connected device Manual Counting p 23 Count the points clicked by user Count Add a point for counting Remove Delete a clicked point Clear Delete all points Adjust p 21 Ad...

Page 37: ...e the distance between two selected points vertically Delete the selected measurement Delete all measurements Layer Merge Layers It shows the merged image of BF and FL channels BF image layer FL FL Green FL Red image layer Features Ruler Measure scale bar manual count etc layer Note Once activating Merge Layers the displayed image is an image processed by setting Opacity and Threshold in the Prefe...

Page 38: ...high value as phototoxicity may occur Moreover it is not recommended to leave the Preview on for a prolonged period as photobleaching may occur Device status It shows the status of connected equipment When IsErrorTriggered lights in red it means that an error has occurred When others light in green it means they are in progress In the case of above image the green lights on IsConnected and IsStand...

Page 39: ...ff Stop the stage on move Vessel Plate panel Vessel type This is the tab where the vessel type is selected Well plate 6 12 24 48 96 wells Dish 35 mm 60 mm 90 mm and T flask 25 cm2 75cm2 can be selected Note The specifications of the well plates are different for each brand On the Celloger Mini Plus Scan app the well plate specifications are set based on a certain brand s specifications As such if ...

Page 40: ...e panel In the case of a well plate it move to the center of the selected well To move to a position other than the center press Ctrl and double click to move to the desired position Scan Pattern Major Order When scanning multiple wells in a well plate the scanning direction can be selected between the Column and Row Direction When scanning multiple wells in a well plate the scanning direction can...

Page 41: ...41 Position Control A B C D ...

Page 42: ...ositions the desired position can be selected and changed from the Scan table C Refer to D on how to change the position FL mode can also be changed in the same way Set Position Set the position you want to scan Current Position Set the current position as the scan position Centers of All Set the center of all wells as the scan position Can only be used when the vessel type selected is a well plat...

Page 43: ...k the selected well and press Ctrl Q C Scan table It shows the scanning order and the corresponding coordinates and Z stacking values Index Well number of the well plate If it is not a well plate all indexes are indicated as A1 X X axis coordinate Y Y axis coordinate Focus Z axis coordinate of BF and FL channels BF channel shows the focus of Z coordinates when Auto Focus function is off None or it...

Page 44: ...refers to taking images at intervals set in Step from a Step bottom to b Step top based on the reference position Focus a and b are the values set in Range refer to Z Stacking of p 46 On the other hand ST c d e f stands for stitching Based on the set position reference position the image is taken with overlapping area set in Overlap while c d e f stands for the range of stitched image left up righ...

Page 45: ...be clicked on for each to apply the changes Click the Apply All button to apply both functions at once FL Focus Offset FL Focus Offset Position of FL channel for time lapse is the location of z axis of FL channel captured while performing the time lapse iamging The value shown is the difference from the location of Z axis captured in the BF channel To capture the FL channel on the same coordinates...

Page 46: ...ile the Step of Z stacking set in Properties is 10 um This means taking images from below 20 2 x 10 um to above 30 3 x 10 um every 10 um based on the Focus of 2 453 As such BF and FL images are taken with the z axis of 2 433 2 443 2 453 2 463 2 473 and 2 483 at x 11 200 and y 32 300 axis At this time 2 and 3 in Util are the values set in the Range Stitching Select whether or not to run the stitchi...

Page 47: ...ely to take the total images of 25 5x5 with overlapping area of 20 as shown in the image below The Reference position 1 is imaged first then the order of imaging is from bottom left 2 to top left 6 in vertical order until it reaches the top right 25 image The images are also stacked and stored in the same order as the imaging order Scan table Properties Stitching Overlap Set position reference pos...

Page 48: ...time of the scan settings Click on to directly open the folder where the file is saved B Schdule Set the schdule for time lapse imaging Time except for End Date and Total Cycle can be selected Click the mouse turn the mouse wheel or click the calender to set the schedule Interval Time Interval between scanning cycles Total Running Time Time lapse execution period Start Date Time lapse start time I...

Page 49: ...indow can be separately floated so that it can fit on the monitor It is possible to change the window by pressing Autohide for floating or hiding The layout is automatically saved upon terminating the program To go back to the original state press Reset Layout in Tool menu before closing the Celloger Mini Plus Scan App When the window disappears delete LayoutCGP XML in the Celloger Mini Plus Scan ...

Page 50: ...p when Celloger Mini Plus Analysis App starts It is for saving and managing parameters set in the Preferences of Tool menu Menu bar Toolbox Layer Mask the area recognized as confluency All other tools and functions are the same as those of Celloger Mini Plus Scan App It is managed by config file instead of a project Other menus are the same as those of Celloger Mini Plus Scan App ...

Page 51: ...ected the image is shown on Display It is possible to analyze images or make a video refer to p 25 34 A Click to load the image file taken with Celloger Min Plus refer to p 26 B Analyze images or make time lapse videos Analyze Calculate the confluence of the files checked on the list Analyze the image of selected light channels on the right refer to p 27 Chart Show the confluency graph Record Make...

Page 52: ... a rigid and level place Operate the device in conditions described in the operating condition Use only the components provided and authorized by Curiosis Inc Ensure the input voltage matches with the device s power supply voltage Check if the power cable is properly grounded to avoid potential electric shock Disconnect the power cable when abnormalities occur Wait about 2 3 minutes for the device...

Page 53: ...wer and wipe dry immediately Make sure that liquid or foreign matters do not enter fan holes or lens holes In case of storage at a room temperature after taking it out of the incubator store it in a dry and well ventilated place Never disassemble the instrument yourself Do not remove any covers or parts that require use of tool to obtain access to moving parts Operators must be trained before bein...

Page 54: ...of objective lens Image is not clear Carefully wipe off the objective lens with cotton swab Eliminate any dust on culture dish and LED lamp Remove any condensation on the lid of the culture dish Problem in saving files Check the unused space of the hard disk Time lapse images become dark and bright To prevent any problem such as shaking of culture dish or inflowing light pay special attention when...

Page 55: ...continues to occur even after performing the above it is recommended not to use the front wells A1 B1 C1 and rear wells as much as possible when using a well plate Note In case of using a 96 well plate using sealing film would prevent condensation and drying of media IX Product specifications Dimension 226 x 358 x 215 mm Weight 5 6kg 12 3lb Objective Lens 4X 10X Imaging modes Brightfield Fluoresce...

Page 56: ...ield Green Fluorescence 10X CRCLG MPBR04 CLG Mini Plus Live cell imaging system Bright Field Red Fluorescence 4X CRCLG MPBR10 CLG Mini Plus Live cell imaging system Bright Field Red Fluorescence 10X CRCLG MPWPS Vessel holder Well plate 6 96 Single CRCLG MPTFS25 Vessel holder T Flask A25cm2 Single CRCLG MPTFD25 Vessel holder T Flask A25cm2 Dual CRCLG MPTFS75 Vessel holder T Flask A75cm2 Single CRCL...

Page 57: ...n 1 1 Device IP setup Celloger Mini Plus is operated based on the connection with PC using wired LAN cable Ethernet connection type Under the Ethernet connection method devices with the same IP address cannot be properly connected In order to connect multiple Celloger Mini Plus units with the same IP address it is necessary to change the IP address of the devices It is possible to change the IP ad...

Page 58: ...ecting 3 Celloger Mini Plus devices and PC PC IP address 192 168 2 200 Device 1 IP address 192 168 2 10 Device 2 IP address 192 168 2 11 Device 3 IP address 192 168 2 20 Note When setting the IP address of the network adapter in Celloger Mini Plus Scan App p 9 only one of two or more devices must be checked in the DHCP Server and the other devices must be unchecked In case of setting the IP manual...

Page 59: ...n Open Celloger Mini Plus Scan App after completing network setup Select the device and connect it Note Devices connected to the App can be identified by choosing one of methods below 1 Check the selected device description 2 Check IP address at the bottom of the program refer to p 49 3 Click Identify and check white LED blinking PC Router Celloger Mini Plus PoE injector PoE injector PoE injector ...

Page 60: ...er IP address 192 168 2 XX and Subnet mask 255 255 255 0 in the blank fields Note Fill in any numbers from 2 254 except 10 in XX fields The PC IP address should be set differently from the device IP address or the network adapter such as a router The default device IP is 192 168 2 10 and to set up the PC IP address enter 192 168 2 XX in XX field input 2 254 except 10 This IP setting guideline may ...

Page 61: ...61 Image descriptions of PC network setting 1 Window 10 Mouse right button click ...

Page 62: ...62 2 Window 11 Properties Properties ...

Page 63: ...63 ...

Page 64: ...he well plate specifications of the brand used in Vessel Editor Select Vessel Editor from the Tool menu Select the type of well plate 6 12 24 48 96 well to be used from the Vessel Editor Selection window Change the specifications Length Width A1LocationX A1LocationY Well Diameter Horizontal Spacing Vertical Spacing of the well plate to be used after entering the Model and Company in the WellPlate ...

Page 65: ...ave the setting values The saved file can be found in the Vessel folder of the Celloger Mini Plus Scan App Re run the Celloger Mini Plus Scan App After checking if there is a well plate of the brand saved in Vessel Type click on it to use it ...

Page 66: ...ssel type As shown in the figure below insert the tip of holder to the area where it faces the LAN port of the device first and mount it Insert the screw into the hole and turn it clockwise to assemble Caution Please be careful when tightening the screw It can fall into the device ...

Page 67: ... 2022 09 15 CR IFU 008 Rev 2 Added contents Grid line Image adjustment Preview record Stitching Chart Revised contents Table of contents Getting started Z stacking Image analysis control Autofocus Cleaning and Maintenance Troubleshooting Appendix B Revised UI Image analysis control ...

Page 68: ...n ro 38 gil Gangnam gu Seoul 06221 South Korea TEL 82 2 508 5237 FAX 82 2 508 5246 Email info curiosis com FACTORY 400 Wonam ro Namsa eub Cheoin gu Yongin si Gyeonggi do 17123 South Korea TEL 82 31 339 6404 FAX 82 31 339 6409 www curiosis com ...

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