GeminiBio ORFLO Moxi V, Руководство пользователя

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Moxi V
™
User Guide
Page 11
1. Solution Conductivity – As the system utilizes an electrical measurement for counting
cells, sizing cells, and metering the fluid volume, cells need to be suspended in a
conductive media. The target solution conductivity would be 0.9% salt solution (e.g.
PBS or equivalent). Deviations of +/- ~50% conductivity can be tolerated by the system.
Most medias that are isotonic to PBS will work.
2. Particle Size and Concentration – For best results tests must be run within the size
and concentration range specifications of the cassette, as specified below.
a. Size Range:
i. Type S+ cassette: 3 - 27 μm diameter (14 – 10,306 fL volume)
ii. MF-M cassette: 4 - 35 μm diameter (14 – 22,449 fL volume)
b. Concentration range:
i. Type S+ Cassettes:
1. Counts: 1 x 10
4
– 1.75 x 10
6
total particles/ml
2. Optimum Fluorescence Sensitivity: < 5 x 10
5
particles/ml
ii. MF-M Cassettes:
1. Counts: 1 x 10
4
– 7.5 x 10
5
total particles/ml
2. Optimum Fluorescence Sensitivity: < 3 x 10
5
particles/ml
Optimal concentration ranges up may vary slightly depending upon your cell
or bead type.
3. Single Cell Suspensions – While the Type S+ and MF-M Cassettes do have a pre-filter
for removing sporadic large particles, cell samples should be prepared as single cell
suspensions to avoid clogging of cassettes. Clusters/aggregates can be broken apart
with mechanical trituration and/or protease dissociation (e.g. Accutase
®
or Accumax
®
).
Larger particles or stubbornly aggregated clusters can be removed with a 40 µm cell
strainer. Standard centrifuge tube strainers can be used for larger volumes or a pipette-
tip FlowMi™ strainer can be used for smaller volume filtering.
4.
As the Moxi V™ uses a 532nm laser with a 561nm/LP filtered PIN diode for detection.
Dye/fluorophore selection should be based accordingly. As the system is intended as a
cell viability system, the main recommended fluophore is Propidium Iodide (PI,
recommended 2µg/ml final PI concentration). Please refer to the
Orflo Viability Staining
Protocol for the staining protocol for viability assessments. For cell counting application,
cells can be run without fluorochromes using any of the assays.
5. Red Blood Cell (RBC) containing samples – For samples with high levels of RBC’s
(e.g. peripheral blood), it is necessary to eliminate/reduce the level of RBC’s in order to
prevent those cells from “masking” the cells of interest. The preferred approaches to
purification of a sample are density-gradient centrifugation (e.g. Ficoll-Paque
®
), magnetic
bead separation, or “pre-plating” (culturing of adherent cells). RBC lysis approaches can
be applied but users should be aware that those approaches have limited efficiency and
can still leave a significant number of intact RBC’s behind.