• 00:00 1.
    Invasion Assay
  • 00:06 2.
    Put transwell chambers in 24-well plate
  • 00:25 3.
    Prepare Matrigel
  • 00:48 3.1
    Add 100 ul Matrigel in the chamber
  • 01:09 4.
    Incubate at 37 C overnight for gelling
  • 01:15 5.
    Add 750 ul culture medium with serum in the lower chamber
  • 01:26 6.
    Place 200 ul cell (2.5x10^5/ml in serum-free medium) in upper chamber
  • 01:51 7.
    Place transwell into lower chamber
  • 02:09 8.
    Incubate at 37 C for 12-16 h
  • 02:13 9.
    Remove medium and wash twice by PBS
  • 02:52 10.
    Fix cells by formaldehyde (3.7% in PBS)
  • 03:17 10.1
    Fix cells at room temp for 2 min
  • 03:20 11.
    Remove formaldehyde and wash twice by PBS
  • 03:55 12.
    Permeabilize cells by 100% methanol
  • 04:16 12.1
    Permeabilization at room temp for 20 min
  • 04:18 13.
    Remove methanol and wash twice by PBS
  • 04:46 14.
    Giemsa stain
  • 05:10 14.1
    Giemsa stain at room temp for 15 min
  • 05:15 14.2
    Remove Giemsa stain and wash twice by PBS
  • 05:43 15.
    Scrape off non-invasive cells with cotton swabs
  • 06:04 16.
    Count invasive cells under a light microscope
  • Index
  • Notes
  • Comment
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Invasion Assay
Duration: 06:17, Browse: 1181, Last Updated: 2018-12-03
    • 00:00 1.
      Invasion Assay
    • 00:06 2.
      Put transwell chambers in 24-well plate
    • 00:25 3.
      Prepare Matrigel
    • 00:48 3.1
      Add 100 ul Matrigel in the chamber
    • 01:09 4.
      Incubate at 37 C overnight for gelling
    • 01:15 5.
      Add 750 ul culture medium with serum in the lower chamber
    • 01:26 6.
      Place 200 ul cell (2.5x10^5/ml in serum-free medium) in upper chamber
    • 01:51 7.
      Place transwell into lower chamber
    • 02:09 8.
      Incubate at 37 C for 12-16 h
    • 02:13 9.
      Remove medium and wash twice by PBS
    • 02:52 10.
      Fix cells by formaldehyde (3.7% in PBS)
    • 03:17 10.1
      Fix cells at room temp for 2 min
    • 03:20 11.
      Remove formaldehyde and wash twice by PBS
    • 03:55 12.
      Permeabilize cells by 100% methanol
    • 04:16 12.1
      Permeabilization at room temp for 20 min
    • 04:18 13.
      Remove methanol and wash twice by PBS
    • 04:46 14.
      Giemsa stain
    • 05:10 14.1
      Giemsa stain at room temp for 15 min
    • 05:15 14.2
      Remove Giemsa stain and wash twice by PBS
    • 05:43 15.
      Scrape off non-invasive cells with cotton swabs
    • 06:04 16.
      Count invasive cells under a light microscope
    Location
    Folder name
    Invasion assay (Abnova 版本)
    Author
    系統管理者
    Branch
    賴亮全教授
    Created
    2018-12-03 11:19:48
    Last Updated
    2018-12-03 11:26:14
    Duration
    06:17