Work out how long to book a cell sorter, and how many cells to bring.
How long to book a sorter, and how many cells to bring. The estimate is arithmetic on numbers
you supply, so it is only as good as your guess at the two things nobody knows before the first
run: what fraction of your cells carry the marker, and how much debris is in the tube.
If you have never run these samples on a cytometer, book an hour for a pilot first.
If your samples are not all alike — different target populations, different cell counts,
some where you need an exact number and some where you want as many as possible — add a
sample type for each and they will all be counted in the same booking.
Instrument figures follow our Cell Sorting Guidelines.
Samples
Instrument
These cells are too large to sort
Write to flowcytometry@chop.edu and we will go through the options for your
cells.
Controls, timings and other assumptions
M/ml
min
min
min
× minimum
Reserve in iLab—
Where the time goes
What to bring per sample unless stated
Before you book
An estimate is not a guarantee. Samples prepared identically can yield different
numbers of sorted cells. Bring more cells than the minimum if you have them — extra
cells let us trade efficiency for speed if necessary.
Single-cell deposition into plates is not covered by this estimate. Filling a
96-well plate with single cells typically takes 3–5 minutes, and considerably longer
when the target events are very rare.
Bring labelled collection tubes with medium or buffer in them — sorted
droplets should land in liquid, not on dry plastic — plus about 10 ml of buffer in
case the cells need diluting, and spare tubes and medium.
Filter the cells through a 35 µm strainer immediately before sorting, to
remove aggregates that can clog the nozzle, and check a drop under a microscope: round,
well dissociated, few aggregates.
Reservations for sorting should be made in
iLab. Choose full-service,
semi-assisted or unassisted under “Pricing details”. Sorters may be reserved in
30 minutes increments.
Estimates only. Setup, event rates and efficiency all vary with the sample; write to
flowcytometry@chop.edu or call 215-590-3402 if the number above looks wrong for
your experiment, and we will work it out with you.
