2 dealing with clogs, 3 cleaning or replacing nozzle tip, 1 unclog option in software – Bio-Rad S3™ Cell Sorter User Manual

Page 98: 2 run sample of cleaner

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88 | S3 and S3e Cell Sorters

Maintenance

10.2 Dealing with Clogs

A clogged or partially clogged nozzle tip can occur while analyzing or sorting and it is usually
a result of an unfiltered sample delivering clumps of cells to the tip. When this happens, the
stream will be unable to spray straight or will be blocked completely. Several things can be
done to remove this clog. These options, in order of severity are listed below. Once complete,
click the startup button and allow the system to verify that the clog has been removed.

10.2.1 Unclog Option in Software

A clog can easily be introduced and can cause multiple issues with the fluidics. Resuspend
cells into a single cell suspension and always use a 40 µm or smaller filter prior to cell sorting
to help avoid clogs. If a clog does occur, use the Unclog button on the Setup and Maintenance
tab toolbar as a first-line action. The S3 and S3e Cell Sorters have the ability to backflush the
nozzle, potentially releasing the clog and flushing it into the waste.

If successful, a couple of debubble cycles are required to remove any air that may have been
pulled into the nozzle body.

If unsuccessful, see Section 10.2.2, Run Sample of Cleaner, or Section 10.4, Swap Nozzle
Tip Wizard.

10.2.2 Run Sample of Cleaner

Running a sample with 70% ethanol or 10% sodium hypochlorite could help free the clog. This
is run through the sample input station. Cleaning solution can dislodge or loosen clogs to free
up the line or the nozzle tip.

If successful, run at least two rinse cycles prior to running any test samples.

If unsuccessful, swap nozzle tip (Section 10.4).

10.3 Cleaning or Replacing Nozzle Tip

The nozzle is the main fluidics component of the system and provides stable sheath flow,
focused sample introduction, and droplet creation and charging. The nozzle tip is fixed at
a 100 µm tip orifice that provides centered sheath stream for laser interrogation. The tip may
be removed to clean or to remove a clog. The nozzle can be accessed through the top, front
sliding cover.

Note: The cover is interlocked and lasers will be turned off if it is opened.

Note: When performing any of the following instructions, use the Swap Nozzle Tip button to
open the wizard, which will assist in swapping or cleaning the tip. The wizard will also walk
through restarting and aligning the system.

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