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Doc.No: 994-2028 v. 1.0 · Issue date: 06-Jan-2020 · Template: 880-0011-79A v.3
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ChemoMetec A/S · Gydevang 43 · 3450 Alleroed · Denmark · support@chemometec.com · www.chemometec.com ·
Within one minute the viability and cell concentration of the sample are displayed. The available results are: Total
(cells/ml), Live (cells/ml), Dead (cells/ml), Viability (%), Diameter (µm), Aggregates (%), Debris Index, Dilution factor and
Status.
If the viability reading is not necessary, then press ‘cancel’ after analysis of the first Via
2
-Cassette™, whereby the total
cell concentration will be provided.
Notes
To ensure robust and reliable results, the cell suspension concentration should be in the range of 5·10
4
to 1·10
7
cells/ml.
If the cell concentration is above 1·10
7
cells/ml dilute with growth media. The diluted cell sample is then counted as
described above.
Lysis
1
will precipitate below 15
°
C. Check buffer for precipitation before use. Re-dissolve any precipitation by warming
to room temperature.
Viability
The percent viability is calculated from:
% Viability =
Ct
−
Cnv
Ct
∗
100%
% Viability
: The Percentage of viable cell in the cell sample
C
t
: The total concentration of cells (Acridine Orange positive cells in the second Via
2
-Cassette™)
C
nv
: The concentration of non-viable cells (DAPI positive cells in the second Via
2
-Cassette™)
Procedure
Adherent or semi-adherent cells are released from the cell culture surface (e.g. by trypsin). For optimal counting precision
use at least 300 µl cell sample and use 1.5 ml micro centrifuge tube, which inner shape fits with the Via
2
-Cassette™ tip.
Materials needed
1.
Cell sample in suspension
2.
Lysis
1
3.
Via
2
-Cassette™
Procedure
Step 1: Determine the total cell concentration from a cell sample mixed 1:1 with
Lysis
1
.
•
The cell suspension is mixed to obtain a homogenous suspension and a sample is mixed 1:1 with
Lysis
1
. E.g., to 100
µl of cell suspension
add 100 μl of
Lysis
1
. Mix by pipetting. Load a cell sample by inserting the tip of the first Via
2
-
Cassette™ into the diluted cell suspension and pressing the piston.
•
Insert the loaded Via
2
-Cassette™ in the NucleoCounter® NC-202™, select the ‘
C&V Lysis-Count’ protocol
, press RUN
and then Select ‘
OK
’.
Step 2: Determine the viability of an untreated cell sample.
•
The cell suspension (
without
Lysis
1
treatment) is mixed again to obtain a homogenous suspension. Draw a cell
sample by inserting the tip of the second Via
2
-Cassette™ into the cell suspension and pressing the piston.
•
When the message box requests it, replace the first Via
2
-Cassette
with the second Via
2
-Cassette™ loaded with the
cell suspension without addition of
Lysis
1
and select ‘
OK
’.
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Summary of Contents for NucleoCounter NC-202
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