ThermoFisher Scientific invitrogen iSort Series, Manual

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iSort
™
Automated Cell Sorter User Guide 31
4. Analyze and Sort
Overview The Analysis and Sort applications of the iSort ™ Automated Cell Sorter have similar
workflows that contain many identical steps. Therefore, they are described together
in this section.
•
Analysis mode (or non-sorting experiment) is designed for quick verification of
sample content and can be performed independently of the Sort workflow. You
can use the Analysis workflow to verify proper sample preparation, to
determine the purity of a sorted cell population, and to verify a successful
sorting process.
•
Sort mode allows you to designate up to two specific cell populations within a
sample, then isolate and collect them. When sorting, the desired populations
are selected by defining sort regions on the density plots that are displayed on
the touchscreen. You can also define a debris gate to exclude a population from
the final sort count.
•
Every time an Analysis or a Sort is completed, the data acquired during the run
are saved in FCS 3.0 format. These files contain the raw data that can be
imported into any standard flow cytometry analysis software (see “Manage
files”, page 46).
Required materials •
Sample (for sample preparation guidelines, see page 32)
•
Sheath fluid (1X PBS, flow cytometry grade)
•
Deionized water
•
Sample tubes, 12 × 75-mm (5-mL capacity)
•
Polypropylene collection tubes, 1.5-mL, 12 × 75-mm (5-mL), or 50-mL tubes in
any combination
WARNING! BIOHAZARD.
When running biological samples for analysis or
sorting, hazardous aerosols may be created depending on the sample type.
To prevent hazardous aerosols from spreading, keep the sorting compartment
door closed as much as possible. Follow all applicable local, state/provincial,
and/or national regulations for the handling and disposal of all
biohazardous substances, including samples, sorted fractions, and waste.
Wear appropriate protective eyewear, clothing, and gloves.