GeminiBio MXG102, Руководство пользователя

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Moxi GO II
™
User Guide
Page 12
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. Fluorescent Labeling/Stains - The Moxi GO II™ uses a 488 nm laser for excitation
with 525/45 nm and either 561 nm/LP or 646nm/LP (second filter is user-swappable)
detection filters. Dye and fluorophore selections should be based accordingly
PMT Filter Compatible Fluorophore Examples (not complete list)
525/45 nm FITC, BB515
®
, Alexa Fluor
®
488, GFP, Calcein, Acridine Orange,
BODIPY 500/510, Rhodamine 110
561nm/LP PE, PE-Cy5, PerCP-Cy5, 7-AAD, PerCP, PI, DsRed
646nm/LP PI, 7-AAD, PE-Cy5
5. Post-Incubation Washes for fluorescent labels – To optimize sensitivity of the
fluorescent signal, it is recommended (for most samples) that the sample be washed 2x
(minimum) with a minimum of 1.5ml staining buffer or PBS to remove the excess (free)
antibody.