SORTING REQUIREMENTS FOR THE
FACSARIA CELL SORTER
1. Book set up time as well as sort time . The set up time will be in the
morning of your sort for 1 hour (recommended between 8 and 9 a.m.). If
you request a sort and the set up time has not been booked; i.e a late
request; you must add 1 hour onto your sort time.
2. Concentrate your cells. We recommend 3-20 million cells/ml.
Although smaller cell numbers than this can be sorted the above
concentration allows the machine to operate at maximum efficiency.
Good starting point is 4 million per ml so we can check if your cells
clump.
3. Book 30 min per ml of sample.
4. Samples to be prepared in no more than 1% serum (if cells require
serum).This prevents frothing and therefore reduces possible
contamination.
5. If possible, Ca/Mg free medium (this helps prevent clumping and
therefore blocking of the machine).
6. Indicate the size of your cells. We have 2 nozzle sizes (70um and
100um); as a rule of thumb the cell diameter should be no more than 1/3
of nozzle diameter. Cells bigger than 20um will need the 100um nozzle.
This will affect the speed of the sort, below are examples of which cells
require which nozzle:Hi Speed (70 um) – Small cells; hi throughput
Med Speed (70um) – Cell lines
Lo Speed (100um) – Large cells.
7. Strain cells through 40um strainer immediately prior to sort.
8. Include relevant antibiotics in your sample and collection tubes.
9. Bring relevant controls (1) Single population sorting requires at least a
negative control but a positive would be desired; (2) Two or more
population sorting requires a negative and a positive for each colour.
This is a basic requirement as we need these controls to set up accurate
parameters and sort gates. NO CONTROLS = NO SORT.
10. Bring two collection tubes containing 2mls of media for each
fragment collected.
11. Ensure all caps and tubes are labelled correctly.
12. BE PUNCTUAL!!!
SORTING OPTIONS
We have various options which can be utilised during a sort ; please
advise at least 24 hours before sorting.
1. Sample Tubes:- we can now sort from 1ml serum tubes; 4ml facs tubes
and 15ml Falcon tubes.
2. Collection:- we can collect into similar tubes or onto plates (6-96
wells)
3. If samples are particularly hazardous we can utilise the Aerosol
Management System of the Aria.
4. If samples require temperature control , the sample chamber can be set
to a choice of temperatures.
5. The collection chamber can also be water cooled if required.