GeminiBio MXG102, Руководство пользователя

Страница: 82 / 93

Moxi GO II
™
User Guide
Page 77
525/45nm filtered emission or 561nm/LP filtered
emission)
Use of Phycoerythrin (PE) with
646nm/LP filter in PMT2 Replace back filter with 561nm/LP filter. PE is
not detectable when using the 646nm/LP filter
Incorrect antibody used.
Make sure the proper antibody clone and target
species is selected for the cells being used.
Fluorescence background is too
high.
Make sure to wash the sample 2x (in a minimum
of 1.5ml of media) to eliminate residual/free
fluorophore.
No fluorophore present
Cell preparation protocol labeling step was
omitted.
Questionable concentration
Concentration of cell/bead sample
is too high or too low
Make sure concentration of cell or bead sample is
within recommended guidelines. Refer to
Specifications section.
Wrong diluent
Cells should be suspended in 0.9% salt solution
(e.g. PBS or equivalent) to ensure proper
conductivity for unit operation and to ensure
proper particle sizing. Water, hypotonic, or
hypertonic solutions are
not acceptable diluents.
Cell clumping
Ensure the cells are in a single-cell suspension.
Break clumps by pipetting up and down with a
standard pipette. Protease treatment can also be
used to dissociate clusters. ORFLO recommends
Accutase
®
or equivalent.
Improper Size Gating
Please refer to the “Managing the Data” or
“Gating the Data” section for instructions on
gating. The noise/debris cluster in the bottom left
corner should be excluded from count analysis.
Questionable cell diameter
Wrong diluent
Cells should be suspended in 0.9% salt solution
(e.g. PBS or equivalent) to ensure proper
conductivity for unit operation and to ensure
proper particle sizing. Water, hypotonic, or
hypertonic solutions are
not acceptable diluents.
Cell clumping
Ensure the cells are in a single-cell suspension.
Break clumps by pipetting up and down with a
standard pipette. Protease treatment can also be
used to dissociate clusters. ORFLO recommends
Accutase
®
or equivalent.
Improper gating
Size measurements are reported as the mean
size for a gated range. If a cell population is not
properly gated, contributions of other cell types,
system noise, or debris particles can contribute to
the mean size calculation.
No clear cell cluster (or
histogram peak on size
histogram)
Improper scaling
This is usually associated with very-low-
concentration samples. For histogram overlay
use finger swipes (left - right) for the fluorescence
histogram and (top – bottom) for the size
histogram. For size histograms (not overlaid),
use the white (black box background) arrows to
adjust the vertical scale to properly scale the
population.