Nexcelom Bioscience

978-327-5340

Image and Quantify Transwell Invasion and Migration Assays using Suspension and Adherent Cells

  1. Image transwell invasion of adherent cells without dissociation of cells from the transwell insert
  2. Directly count the number of invading/migrating cells
  3. Fluorescently label and image invading cells

Perform and Quantify Transwell Invasion Assay by Direct Cell Count

transwell illustration

Suspension cells located inside the insert migrate through the porous membrane toward the chemoattractant from the bottom plate. Celigo is used to automatically count migrated cells in the bottom plate.

Image and Count the Number of Cells in a Transwell Invasion Assay

Protocol

  1. Three different adherent cancer cell lines were seeded into a transwell insert with or without treatment to stimulate migration.
  2. 24 hrs after seeding, the cells were fixed in the transwell insert with formaldehyde
  3. Cells were then washed with water to remove any formaldehyde
  4. Cells which had not migrated through the membrane were removed from the top of the transwell insert
  5. Cells were stained with DAPI/Triton X-100 solution for approximately 10-15 minutes
  6. Remove transwell insert and wash with PBS
  7. Add 600uL of PBS to a clean well of a 24-well BD Falcon plate
  8. Place washed transwell insert into well and image on Celigo using bright field and blue channels

Bright field and DAPI images of transwell invasion of adherent cancer cells

transwell invasion of adherent cancer cells

Overlay of bright field and DAPI

Overlay of bright field and DAPI

High resolution bright field images of the transwell membrane

high resolution bright field images of the transwell membrane

Thumb-nail pictures and cell numbers are automatically displayed upon the completion of the cell counts

Acquisition of high resolution DAPI and bright field images of the transwell membrane surface in a 24 well plate format took ~ 3 minutes

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