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Find answers to frequently asked questions on chemotaxis assays.

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Types of Chemotaxis Assays

Statistics

Typically, three to five repeat experiments are sufficient to create significant data from chemotaxis and control experiments. To achieve this, each experiment should contain data from 20 - 40 single cells, which is possible using low-magnification microscopy objective lenses, such as 5 x or 10 x.


Typical results of a chemotaxis experiment with HT-1080 cells migrating towards fetal calf serum. The FMI (forward migration index) values parallel (║) and perpendicular (┴) to the gradient are shown. Significant chemotaxis is only observable parallel to the gradient in the +/- experiment. All other values are not significantly different from 0.


Experimental Endpoints

  • Chemotactic behavior of cells and mutants (knockout / knockdown)
  • Chemotactic potential of substances
  • Effect of chemotaxis inhibitors / enhancers
  • Distinguishing between chemotaxis and chemokinesis (enhanced migration)
  • Molecular mechanisms during chemotaxis, visualized by fluorescence microscopy
  • Investigation of signal transduction or cytoskeletal effects during

Experimental Examples

All examples are carried out in the µ-Slide Chemotaxis 2D or µ-Slide Chemotaxis 3D, respectively.

HUVEC Chemotaxis Inhibition

Endothelial cells, which tend to migrate in a collective manner, can be tested on compounds that inhibit chemotaxis functionality, even when there is a gradient present (e.g., compounds suppressing cytoskeleton dynamics, or receptor antagonists).

Leukocytes

Fast leukocytes, such as dendritic cells or T-cells, can be visualized well during chemotaxis, when inside gel matrices like collagen. This is especially true when using high-resolution fluorescence microscopy, which shows changes in the cytoskeleton during immune response and 3D interstitial migration.

Cancer Cells

Chemotaxis, described as the directed motion towards a chemoattractant, can be distinguished from chemokinesis, which is known as an enhanced migration effect. Both effects can be analyzed independently of each other. In the example, the chosen chemoattractant induces both chemotaxis and chemokinesis.

Cell-to-Cell Chemotaxis

The ibidi μ-Slide Chemotaxis 3D can additionally be used for various cell-to-cell chemotaxis assays. For example, cells brought inside the large reservoirs can be used as a chemoattractant producer.