BD FACSAria Manuale rapido - Pagina 2

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8. Close the hood and ensure it snaps into place. This will allow the laser interlock to

disengage and allow laser light to pass through the flow cell.

Setting up the break off point

1. . Make sure that the stream appears in the Break off Window.

a. No stream, dripping or spraying: remove nozzle tip. Sonicate the nozzle,

wipe out the flow cell and sort chamber.

b. Unsteady stream: shut off the stream, turn it on again. If problem persist,

remove the nozzle tip, sonicate, and put it back on again.

c. Stream obscured by grey area: sheath fluid in front of droplet camera. Turn

off the stream, remove the nozzle tip and the key, dry everything out, and

clean the chamber with kimwipe.1

2. Monitor the droplet formation

Gap

Drop 1

Satellite

droplets

70um tip

85/100/135um tip

3. The leftmost figure above shows fast converging droplets wherein the satellites

attach to the top of their droplet. Fast converging droplets are required to use the

70um tip and are preferred for the 100um tip. Slow converging droplets, shown in

the rightmost figure, are never preferred but can be used for the 100um tip. If you

observe slow converging droplets while using the 70um, remove and sonicate the tip

as slow converging droplets can never be used with the 70um tip.

Gap (wide band): distance between the

unbroken stream and the first unattached

droplet

Drop1 (thin band): position of the first

unattached droplet

70/85um tip: satellites attach to the droplet

on top, the gap is located at between the

above satellite droplet and the following

droplet.

85/100/130um tip: Satellites attach the

droplet below, gap is located just on top of the

last attach droplet and its following satellite

droplet