Flow cytometry facility // Al. Fleming BSRC

CELL SORTING GUIDELINES
-	Please get in contact to arrange for an appointment and discuss your experiment. It is strongly advised to refer to the Fluorochrome Reference Chart (BD FACS Vantage SE II) for fluorochrome compatibility of the instrument and if possible, please bring along a FACS analysis of your experiment.
-	Samples, must accompanied with a filled copy of the Experiment Description Sheet as well as a FACS analysis of your experiment to be used as reference.
-	There are some essential requirements for you experiment to be carried out efficiently.
	-	Clean cell preparations. Cell preparations containing debris and aggregates will affect the experiment outcome and probably clog the flow line of the instrument. Unclogging can become very tedious and time consuming. To avoid such a situation please filter your samples with nylon mesh (70μm) before you bring them.
	-	Compensation Controls. All experiment will require controls for the proper set up of the instrument and outcome of your experiment. The basic idea behind controls is described in the following example.
		-	Sample: mouse spleenocytes stained with FITC, PE, PE-Cy5.5
		-	Controls:	Tube 1: unstained cells Tube 2: single-stained FITC Tube 3: single-stained PE Tube 4: single-stained PE-Cy5.5
	-	Sample cell densities and volumes. All samples should be brought inFalcon 12x75mm polystyrene tubes.
			For controls: tubes must contain 500-600 μl at a minimum concentration of 1.0x10⁶ cells/ml
			For sorting: tubes must contain 3.5ml maximum at a concentration of 2.5x10⁶ cells/ml.
	-	The total number of cells and volume of starting cell preparations as well as the numbers of cells sorted depend on the nature of the experiment and instrument capability and they will be determined following discussion.