Chemotaxis

5 posts / 0 new
Last post
golfer1
golfer1's picture
Chemotaxis

Hi all,
I'm a new person here but was just wondering if anybody had a particular preference for a technique to measure chemoaxis i.e. polycarbonate filters, gels, chambers etc.
Thanks

Kvachhani
Kvachhani's picture
hi,

hi,
i think you are talking about chemtaxis, well it is very difficult to measure but below site may be useful to you.
http://www.copewithcytokines.de/cope.cgi?key=Chemotaxis
http://www.springerprotocols.com/Abstract/doi/10.1385/0-89603-408-9:233
regards.
 

The FFM
The FFM's picture
You can find a Chemotaxis
heehawmcduff
heehawmcduff's picture
Hi there,

Hi there,
I have used two different types of chemotaxis assay:
1. The transwell migration assay where cells are placed in the apical compartment of transwell supports separated from the chemotactic stimulus which is placed in the lower compartment.  The transwells use polycarbonate filters with varying pore sizes for each cell type that you wish to use.  For example, 3um pore size when studying the migration of neutrophils and 5um pore size for macrophages.  After the transwells have been incubated for 1-6 hours (depending on cell type, stimulus etc), the membranes from each transwell can be removed and the cells on the lower surface stained by a variety of methods to enable counting of the cells. 
I found this assay to be pretty reliable and repeatable and would certainly recommend it.  One of the advantages is that, following fluorescent staining, you can mount the membrane on a glass slide and examine the cells that have migrated by confocal microscopy as well as normal light microscopy.
2. I have also used a 96-well chemotaxis chamber (neuroprobe) which is reusable and is supplied with filters through which the cells migrate.  I wasn't really impressed with this chamber as it had a propensity to leak (a problem which other people in another lab with the same chamber type also reported).  However, that said, when it did work, the results were repeatable and made sense i.e. cells migrated towards positive stimuli but not towards negative stimuli.
 
You may also find this paper of interest.  It was by Peter Wilkinson in Glasgow, UK and he has a great amount of experience with measuring cell locomotion and identifying the various types of migration that can occur.  This paper describes many different assays of leukocyte migration and compares and identifies the most appropriate ones to use.
http://www.ncbi.nlm.nih.gov/pubmed/9760220?ordinalpos=1&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum
 
Hope you find it helpful. 
Best wishes

The FFM
The FFM's picture
Kvachhani wrote:

Kvachhani wrote:

hi,
i think you are talking about chemtaxis, well it is very difficult to measure but below site may be useful to you.
http://www.copewithcytokines.de/cope.cgi?key=Chemotaxis
http://www.springerprotocols.com/Abstract/doi/10.1385/0-89603-408-9:233
regards.
 

 
The springer protocols are a useful resource but only if you or your institute has a subscription.