Miltenyi Biotec MACSQuant Tyto Manual Del Usuario página 46

The input filter is clogged
Sort and event rate decrease immediately during sort run, but transit time of cells is correct. This can occur if:
The sample contains high amount of debris or cell clusters. Clean up the sample (filtration, density gradient,
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dead cell removal).
Deactivated or non-functional mixer (sedimentation of cells). Check the mixer. Use a new cartridge if necessary.
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False negative fluorescent population
If primary trigger and/or selected cell speed channels are wrong or the transit event rate is too low, dot plots
show false negative populations but the transit time says 40 µs for regular speed cartridges or 20 µs for high-
speed cartridges. For an overwiev of
Examine trigger and speed channel selection.
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Examine transit event rate, check if trigger for 2nd transit channel is too high.
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Display time-of-flight histogram (HDR-TOF) (refer to MACSQuantify Tyto Software User manual).
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Cartridge
5
MACSQuant Tyto Cartridge
MACSQuant Tyto Cartridge HS
MACS GMP Tyto Cartridge
Table 5.1: Overview of cartridge types.
Low yield
A low yield after sort can have multiple reasons:
Timing of detection and sort valve are incorrect.
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Flow control settings are incorrect and thus the transit time is incorrect (Figure 5.5).
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Detector setting is too low.
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Arrival windows are too small or too large.
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Valve timing is incorrect.
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Alignment of cartridge is incorrect.
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Event and/or sort rate is too high.
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An air bubble is in the channel or the sort chamber.
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Do one or all of the following:
Check trigger channel and speed channel selection.
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Select event transit time or adjust sample pressure manually to reach a transit time of 40 µs for regular speed
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cartridges or 20 µs for high-speed cartridges (Figure 5.5). For an overwiev of Cartridge types refer to Overview
of cartridge types. above.
Check setup of detectors.
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Cartridge types refer to Table 5.1 . Do one or all of the following:
Ordern no.
130-104-791 (8 pieces)
130-106-088 (24 pieces)
130-121-549 (8 pieces)
130-121-551 (24 pieces)
170-076-011 (1 piece)
Regular speed High-speed
yes
no
yes
(HS)
no
yes
no