Cell Migration Assay Cell Émigré
Cell Migration Assay Cell Émigré
The Cell Migration Assay, Cell Émigré, is a quick & quantitative assay for determining linear cell migration rate. It is designed for use with attachment-dependent mammalian cells.
This assay measures migration within an array of linear microchannels in a microfluidic chip. The assay kit comprises a set of single-use microfluidic migration chips together with support reagents and protocol. Video recording/computer image analysis is not required and migration can be quantified at any time point during an assay.
Each Cell Émigré assay chip comprises three identical migration arrays. Each array contains ten microchannels. A sample of cells in fluid suspension is applied via the pipette access port. Following cell attachment the microchannel wall guides each cell along a linear migration path by contact guidance. Quantification requires measurement of a cells linear cell displacement from the ‘START’ position, via an integrated scale.
- Cell Émigré is designed to measure the linear migration rate of attachment-dependent mammalian cells.
- Migration rate responses can be readily compared across different cell types. The assay provides a rapid means of evaluating and comparing the effect of chemical or biological treatments such as chemical agents, proteins, peptides, antibodies or genetically modified cells.
1. AN EASY-TO-USE ASSAY
The end-user supplies a cell sample (of freshly detached cells in fluid suspension) to a microfluidic ‘chip’. The chip then automatically distributes cells to microchannel arrays – ready for attachment & migration. No pumps or external fluid connections are required; each chip contains a reservoir for culture medium which is filled during the initial cell addition step.
2. MEASURES THE RATE OF LINEAR CELL MIGRATION – NO MORE GOING IN CIRCLES!
Existing 2D-migration assays often attempt to measure ‘random-walk’ cell migration – a challenging task! Cell Émigré takes the innovative approach of quantifying migration within an array of linear microchannels. Each cell under assay is constrained to a linear migration path through contact-guided migration along a microchannel wall.
3. RAPID MIGRATION QUANTIFICATION
Cells begin migrating within Cell Émigré aligned to a common START position; a cells displacement from this position indicates its linear migration distance. There is no need for video recording or computer image analysis.
Each microchannel in the array contains a parallel measurement scale calibrated in microns. Cell migration can be rapidly & easily quantified (to the nearest 50μm) by visually comparing the alignment of a cell within the microchannel with this integrated measurement scale (using x40-x100 microscopic examination). Migration can be quantified at any time point during an assay.
Results may be expressed as a migration rate: microns per hour (μm/h).
4. REDUCED REAGENT CONSUMPTION
Achieved through the use of microfluidic technology. A single chip containing 3x assays consumes <350μl of fluid when loaded. This reduces costs in terms of cells and reagents.
Stimulation of linear migration in HT1080 (ATCC® CCL-121) and CHO (ECACC 85050302) cell lines using Foetal Calf Serum (FCS) at a concentration of 0-15%.
Inhibition of linear migration in HT1080 (ATCC® CCL-121) and CHO (ECACC 85050302) cell lines using Cytochalasin D at a concentration of 0-100nM.
Q. Why am I getting air bubbles in my microchannels?
A1. Sudden temperature increases may cause dissolved gas to come out of solution, forming bubbles within microchannels. To prevent this – allow the Primer and culture medium solutions and Cell Émigré assay chips to come to room temperature prior use.
A2. Air bubble entrapment can be caused by partial evaporation of fluid from the supply channel during chip loading. This is more likely to happen in laboratory environments above room temperature (>23 °C). To prevent evaporation we recommend that reservoir fluid removal and replacement occurs as promptly as possible by removing and replacing fluid one reservoir at a time. Do not process multiple reservoirs in batches.
Q. How long does the Cell Émigré assay take to perform?
A. The assay time will depend on the particular cell line being used. We recommend allowing the cells to migrate a minimum of 200 μm before attempting to calculate a migration rate. Rapidly migrating mammalian cell lines will achieve this is ~2 hours, other cell lines may take 15 hours to achieve the same migration distance.
Q. Can I apply coatings to the microfluidic channels of Cell Émigré?
A. Customer-provided coatings may be used in Cell Émigré. Coatings must be of sufficiently low viscosity to flow through the microchannels after being applied via the pipette access ports. A general coating protocol may be downloaded below:
Results from a Laminin-coating experiment may be found on P12 of the assay manual (available for download below).
Q. Can I re-use Cell Émigré chips?
A.Cell Émigré chips are single use only, for the following reasons:
- Not only is it difficult to clean the microchannels from cells and adsorbed protein, it is also impossible to verify if suitable cleaning has taken place.
- Each chip is constructed from a bio-compatible silicone top. Over the course of an assay the silicone can absorb compounds from the culture medium and the cells, as well as any compounds that have be supplied to modulate migration. If chips are reused, these compounds could leech into solution and influence any future experiments.