eBook: Optimizing Cell Culture Techniques - 10

be aspirated and then dispensed into the 96 well
plate. The cell culture medium and the cells will
be mixed for 30 cycles. When this step is finished,
the pipette aspirates 120 μl of cell suspension and
alerts the user to change the three stage position,
sliding it into position 1. The VIAFLO 384 dispenses
40 μl of cell suspension to a new 96 well plate,
then prompts the user to change plate. This step
will be repeated twice, resulting in the cells being
split into three new plates.
Tip: In this example, the ES-E14TG2 mouse embryonic
stem cells were split in three. This can easily be
set up differently for other cell lines by changing
the aspiration and dispensing volumes.
Experimental results (Friedrich Miescher Institute
for Biomedical Research) showed consistency between
replicates of mouse embryonic cell samples
with the same confluency. The mixing step was
very efficient, allowing single cells to be seen under
the microscope (Figure 3c).
Remark
Partial plates
Programs can be easily adapted to a different number
of samples. The VIAFLO 384 is able to work with
any number of tips loaded, giving you the benefit
of simultaneous and accurate dispensing of a
smaller number of samples.
Conclusion
* The VIAFLO 384 offers a simple, accurate,
reproducible, and fast pipetting solution for
cell washing, transfer and seeding.
* Prolonged manual pipetting tasks can
lead to repetitive strain injury. This can be
avoided by automating the cell culture
steps with the VIAFLO 384, maximizing
hands-free time.
* Optimized electronic mixing and
perfect pipetting height control achieve
homogenous cell suspensions while
minimizing the chance of cell shearing
during passaging and splitting.
Figure 3. Microscopy image showing the different stages of the
splitting protocol for four different wells. Pictures of mouse embryonic
stem cells were taken a) after washing the cells with PBS,
b) after the incubation with accutase, c) after splitting the cells,
and d) 24 hours after splitting the cells. Wells 1 and 2 showed
lower confluence, while wells 3 and 4 had higher confluence. An
Axiovert 40 CFL (Zeiss) microscope was used with 2.5x magnification.
(Photos courtesy of Julie Cramard, Friedrich Miescher Institute
for Biomedical Research, Basel, Switzerland.)
* The VIAFLO 384 with a 96 channel pipetting
head was used in this example, but the
VIFALO 96 and VIAFLO 384 base units have
easily interchangeable heads to allow 24, 96
and 384 channel pipetting, providing the
right tool to match your throughput. The MINI
96 offers another option for this application,
but in a much smaller and simpler package.
10
eBook: Optimizing Cell Culture Techniques

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