monocyte chemotaxis assay - (Jul/07/2008 )
Hi all! I want to measure efect of some protein toward monocyte migration. I have read that chemotaxis of monocyte can be assess by use boyden chamber, but I dont find the principal of this methode and its prosedure. Anyone can help me to give more information about that and the refference?
Dear Fiat29,
The principle of the Chemotaxis assay is simple. The Boyden chamber is placed/suspended in a 12/24 well plate. The chemoatttractant is placed under the Boyden chamber in the plate BUT in contact with the Boyden chamber. The freshly isolated monocytes are then placed in the Boyden chamber. The bottom of the Boyden chamber has a filter which over the course of the experiment is invaded by the monocytes in response to the chemoattractant. The filter is removed and the infiltration of the filter can be assesesed microscopically. The stronger the chemoattractant, the further the monocytes walk through the filter
Hope this is useful
Kindest regards
Rhombus
Dear Fiat29,
The principle of the Chemotaxis assay is simple. The Boyden chamber is placed/suspended in a 12/24 well plate. The chemoatttractant is placed under the Boyden chamber in the plate BUT in contact with the Boyden chamber. The freshly isolated monocytes are then placed in the Boyden chamber. The bottom of the Boyden chamber has a filter which over the course of the experiment is invaded by the monocytes in response to the chemoattractant. The filter is removed and the infiltration of the filter can be assesesed microscopically. The stronger the chemoattractant, the further the monocytes walk through the filter
Hope this is useful
Kindest regards
Rhombus
Ok, thanks a lot for your response, It make me more understand and interest to do this methode, can you give me an appropriate refference about it from journal or web address? Thanks Rombus
Protocol & info: http://www.neuroprobe.com/products/chemo_tx.html
Starting point: "Analysis methods of human cell migration" Experimental Cell Research, Volume 307, Issue 2, 15 July 2005, Pages 418-426
Important note: the Boyden Chamber only offers an end-point result of chemotaxis.
Alternatives when live imaging is needed are the Dunn chamber or microslide (Ibidi corp.). If you are using the Boyden-chamber, look into quantification methods, there are a few available, each with it's own pro's and con's.