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Cell Line Optimization


                     Cell Colony Picking                                                                                                CELL PICKER





                               Cell Colony Picking Method Used to Automate                                                                                 benefits

                                 Picking Operations for Cell Genome Editing                                                                                                                                                                  Cell Line Optimization
                                                                                                 click here                                     •  Automation of manual steps enables reliable picking operations.
                                                                                                                                                •  Operations can be recorded easily.
                     Operating Principle and Features                                                                                           •  The compact space-saving design is ideal for installation in cramped laboratories.
                                                                              Table 1   Adherent Cell Cultivation Parameters
                     CELL PICKER has a technology to aspirate and discharge liquids.     Cells:  HCT116 cell line (human colon cancer cells)
                     After visually deciding the target cell colony, a button is pressed to   McCoy’s 5A medium
                     automatically attach a pipette tip to the end of the nozzle and reliably   Culture medium:  with 10 % FBS, 2 mM glutamine added                                                                                         Culture
                     move the tip close to the target cell colony. When the tip scrapes   Coating:  Gelatin solution                                                                                      Reduces work by enabling cell
                     off the cell colony, the measuring pump simultaneously activates to   Cultivation parameters:  Let stand at 37 °C and 5 % CO2                                                        observations and instrument
                     aspirate and then discharge the cell colony and a small amount of the                                                     Hand shakes                                                operations via a tablet computer!
                     medium. Using a tablet computer for observations and operations can   Genome-edited cells are
                     reduce the amount of work involved in operations.  seeded to form colonies.
                     Procedure and Cultivation Parameters

                                                  6
                     A 6-well plate was seeded with 1×10  to 1×10  cells/well of human
                                            4
                     colon cancer cells (HCT116 adherent cell line). After cultivating the cells                                                                                                                                             Purification
                     for six days, the CELL PICKER was used to pick cells and seed a 96-well
                     plate. Then the same cultivation parameters were used to cultivate the
                     seeded cells for six days, after which the cell adhesion and proliferation
                     were checked. The operation process flow is illustrated in Fig. 1.                                                               Button-operated
                                                                                                                                                      tip
                     Results                                          Cell colonies are picked with
                                                                      CELL PICKER.
                     Cell colonies before and after picking are shown in Fig. 2. The picked                                                                    improves
                     cells after cell proliferation are shown in Fig. 3.                                                                                       reliability!!
                     100 % of the seeded wells produced adherent cell cultures.  Clones are established.
                     This example confirmed that cells can be picked and seeded without                                                                                                                                                      Characterization
                     causing cell damage.                                          Fig. 1   Workflow image
                     Application Examples                               Before picking   After picking

                     • Picking for establishing iPS cells
                      This confirmed that iPS cell colonies can be cultivated continuously
                      while maintaining their undifferentiated state.
                     • Collecting floating cell clusters (Spheroids)
                      The system can also be used for the purpose of collecting spheroids   Fig. 2   Cell colonies before and after picking                                                                                                  Quality Control
                      created by 3D cell culturing.
                      It can pick a single spheroid from among multiple spheroids in an   Day1  Day4  Day5                              Specifications
                      HEK293 cell line (400 to 500 μm) (Fig. 4).
                                                                                                                                         Instrument            CELL PICKER
                                                                                                                                         Microscope (recommended)  Olympus CKX53
                                                                                                                                         Pipette tip volume    200 µL
                                                                                                                                         Recommended pipette tip  QSP (Thermo Fisher Scientific)
                                                                       Note: These colonies are different than shown in “Cell colonies before and after picking.”  TW110-96RNS-Q
                                                                                                      Scale bar: 200 µm
                                                                                Fig. 3   Cell proliferation after picking                Cell culture vessels  6 well plate: FALCON_353046 / IWAKI_3810-006                                  Pharmacokinetics
                                                                                                                                                               6 cm dish: FALCON_353002 / IWAKI_3010-060
                                                                       Before picking    After picking                                                         10 cm dish: FALCON_353003 / IWAKI_3020-100
                                                                                                                                         Suction amount        Picking mode: 5/10/15 µL
                                                                                                                                                               Removal mode: 5 µL
                                                                                                                                         Dimensions            W 280 mm × D 350 mm × H 400 mm (not included microscope)
                                                                                                                                         Weight                Approx. 8.5 kg                                                                Others
                                                                                                                                         Power supply          100 to 240 V AC, Frequency: 50/60 Hz, Power consumption: 75 VA
                                                                            The white dashed circles in the images are not shown in the software.
                                                                             Note: The indicated operations require special operating software.  Operating environment  Temperature: 10 to 35 ˚C     Humidity: 20 to 85 % RH
                                                                               Fig. 4   Before and after picking spheroids              Note: A table PC for operation is required separately.


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