Cell Migration & Invasion Assays

Metastasis is the cumulative result of multiple changes in tumor cells and their microenvironment that enables cellular migration and invasion into healthy host tissue. As proliferating neoplastic cells attempt to escape the primary tumor site, local cell adhesion and invasion of the surrounding tissue must occur1. Prior to penetrating the blood vessel endothelium and gaining access to the blood stream, cancer cells must invade local tissues by degrading ECM protein components and ultimately, transverse the basement membrane2. Once in circulation, these cells can form metastatic colonies at secondary locations.

Tumor metastasis is a multistep process involving cancer cell adhesion, cell migration and invasion into neighboring tissues and vasculature

Figure 1. Tumor metastasis is a multistep process involving cancer cell adhesion, cell migration and invasion into neighboring tissues and vasculature.

The Boyden Chamber Assay

The most widely accepted cell migration technique is the Boyden Chamber assay3. The classic transwell migration assay system uses a hollow plastic chamber, sealed at one end with a porous membrane. This chamber is suspended over a larger well which may contain medium and/or chemoattractants. Cells are placed inside the Chamber and allowed to migrate through the pores, to the other side of the membrane. Migratory cells are then stained and counted.

 The Boyden Chamber Assay Protocol

Figure 2. The Boyden Chamber Assay Protocol. Cells are allowed to migrate through a cell monolayer or ECM protein mixture which have been seeded onto a semi-permeable membrane cell culture insert with chemoattractants added below the membrane. Migrated cells can then be quantified by staining cells with DNA dyes such as Calcein-AM or CyQUANT GR Dyes.

Cell Migration & Invasion Kits

Millipore’s QCM™ cell migration and invasion assays provide a quick and efficient system for the quantitative determination of various factors on cell migration, adhesion and invasion including screening of pharmacological agents, evaluation of integrins, chemoattractant or other adhesion receptors responsible for cell migration. The kits have provided researchers with consistent results for over a decade and have been published in over 4000 Publications.

Cell Migration Assays: Enables convenient and sensitive quantification of in vitro cell migration towards a chemical concentration gradient (chemotaxis) or ECM protein gradient (haptotaxis).

Cell Invasion Assays: Enables convenient and sensitive quantification of in vitro cell invasion through a basement membrane ECM protein or a layer of cells such as endothelial cells.

How to select the appropriate pore size for your cells:

  • 3 μm pore size is appropriate for leukocyte or lymphocyte migration.
  • 5 µm pore size is appropriate for a subset of fibroblast cells or cancer cells such as NIH-3T3 and MDA-MAB 231 cells. Also suitable for monocytes and macrophages.
  • 8 μm pore size is appropriate for most cell types. This pore size supports optimal migration for most epithelial and fibroblast cells. Note - the 8 μm pore size is not appropriate for lymphocyte migration experiments.

Cell Migration Assays

Description Pore Size Plate Format ECM Coating Detection No. of Tests Product No.
Chemotaxis Cell Migration Assays 8 µm 24-well
None Colorimetric 24 ECM508
    24-well   Fluorometric 24 ECM509
    96-well   Fluorometric 96 ECM510
  5 µm 24-well   Colorimetric 24 ECM506
    24-well   Fluorometric 24 ECM507
    96-well   Fluorometric 96 ECM512
  3µm 24-well   Colorimetric 24 ECM504
    24-well   Fluorometric 24 ECM505
    96-well   Fluorometric 96 ECM515
Haptotaxis Cell Migration Assays 8 µm 24-well Fibronectin Colorimetric 24 ECM580
    24-well Collagen I Fluorometric 24 ECM582
  5 µm 24-well Laminin Colorimetric 24 ECM220
    24-well   Fluorometric 24 ECM221

Cell Invasion Assays

Description Pore Size Plate Format ECM Coating Detection No. of Tests Product No.
Cell Invasion Assays 8 µm 24-well ECMatrix™ Colorimetric 12 ECM550
    24-well   Colorimetric 24 ECM554
    96-well   Colorimetric 96 ECM555
    24-well Collagen I Colorimetric 24 ECM551
    24-well   Fluorometric 24 ECM552
    96-well   Fluorometric 96 ECM556
Endothelial Cell Migration Assays 3 µm 24-well Fibronectin Colorimetric 24 ECM200
    24-well   Fluorometric 24 ECM201
Endothelial Cell Invasion Assays   24-well ECMatrix™ Colorimetric 24 ECM210
    24-well   Fluorometric 24 ECM211
Leukocyte Transendothelial Migration 3 µm 24-well Fibronectin Colorimetric 24 ECM557
Tumor Cell Transendothelial Migration 8 µm 24-well   Colorimetric 24 ECM558
QCM™ Invadopodia Gelatin Degradation Assay (Green) NA NA FITC-Gelatin* Fluorometric 32 ECM670
QCM™ Invadopodia Gelatin Degradation Assay (Red)     Cy3-Gelatin* Fluorometric 32 ECM671

Microfluidic Migration Device

Millicell® µ-Migration Assay Kit overcomes the limitations of traditional multiwell migration assays. The innovative design of the µ-Migration Slide promotes a stable diffusion-generated concentration gradient that is consistently linear and lasts for more than 48 hours. Made from a plastic with high optical qualities similar to those of glass, the µ-Migration Slide is specially designed for video microscopy assays. At specific time intervals, images of the observation area can be acquired, allowing real-time monitoring and quantitative measurements of cell migration.

Live Cell Migration of HT1080 cells

Figure 3. Live Cell Migration of HT1080 cells. The effect of serum concentrations (0% or 10%) on the migration propensity of HT1080 cells on collagen-coated surface. The results show that the majority of the cells have directed migration towards 10% FCS gradient (Black).

In Vitro Scratch Assay

The scratch assay is a popular method for the study of cell migration4. Scaling up this technique has proved challenging, however, making biochemical analysis of the molecular events mediating wound repair difficult. The Cell Comb™ scratch assay addresses the need for a simple tool able to create multiple scratch wounds in a higher-throughput manner.

Read our Application Note in Nature

Product No. Product Description
17-10191 Cell Comb™ Scratch Assay

CellComb scratch assay

Figure 4. Using the CellComb scratch assay, monolayers of NIH3T3 cells were scratched/wounded in a one-direction (left) or two-direction (right) pattern. Over time, migrating cells will fill in the scratched sections and quantified using simple cell counting. Live cell imaging on migrating cells is also possible using the scratch migration assay.

ECM Proteins

Extracellular matrix (ECM) proteins are produced intracellularly and are subsequently secreted into the surrounding cellular medium, actively regulating a diverse range of cell functions including cell adhesion, differentiation, proliferation, migration, invasion and survival. A primary utility of ECMs in in vitro culture is to promote cellular adhesion while maintaining cell viability and maximizing cell proliferation for downstream cell-based applications.


  1. Friedl P, Alexander S. Cancer invasion and the microenvironment: plasticity and reciprocity. Cell. 2011 Nov 23;147(5):992-1009.
  2. Zena Werb et. Al. Extracellular Matrix Degradation and Remodeling in Development and Disease. Cold Spring Harb Perspect Biol. 2011 Dec; 3(12): a005058.
  3. Chen HC. Boyden chamber assay. Methods Mol Biol. 2005;294:15-22.
  4. Guan JL. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc. 2007;2(2):329-33.