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Figure 1.
Milling Mashing Lautering Boiling Whirlpooling
Malted The Brewing Process.
Barley Accurate counting of yeast
Hops cells is important during the
H20
pitching and yeast sampling
steps of the brewing
process.
Yeast Cooling
Packaging
and Yeast
Distribution
Yeast Sampling
Filter Cover
Results
Handle Grip Samples of varying concentrations from a single
overnight culture were used to assess the feasibility of
counting yeast with a Scepter cell counter fitted with
40 m aperture sensors. For all concentrations tested,
Tip Attachment Scepter Sensor yeast samples yielded interpretable histograms that
Port
could be gated and used to calculate yeast size and
concentration. Examples of these histograms are shown
in Figure 3.
Comparative performance
Orifice To assess the range and linearity of Scepter cell
Microchannel
counters performance, we plotted the measured
concentration values (mean of 4 replicates) versus
the corresponding theoretical concentrations across
Front Back
the dilution range. Identical samples were tested in
2
R2=0.995
(cells/mL)
8.0E+05
1.6E+06
4.0E+05
(cells/mL)
0.0E+00 4.0E+05 8.0E+05 1.2E+06 1.6E+06
8.0E+05 Theoretical Cell Concentration
Count
1.5e6/mL
250
(cells/mL)
1.0e6/mL
7.5e5/mL 4.0E+05 Theoretical Cell Concentration
150
5.0e4/mL
Theoretical Cell Concentration
3 6 9 12 15 18 (cells/mL)
Diameter (m)
Theoretical Cell Concentration
Z2 Coulter Counter
Figure 4.
400
Scepter - 40 m Sensors
For Scepter cell counter and Z2 Coulter Counter, measured yeast cell concentrations are
plotted against the theoretical concentrations. The solid gray line represents the theoretical
320
values. For each device, the dotted lines represent best linear fit to the seven data points.
7.5e5/mL
Both platforms show a loss of linearity and counting accuracy with the increase in cell
concentration.
240
Count
160
1.25e5/mL
80
3 6 9 12 15 18
Diameter (m) 50,000 53,950 2.1 53,893 1.7
125,000 129,175 1.4 132,708 1.1
250,000 252,375 3.8 257,115 0.5
Figure 3.
Scepter Software Pro permits yeast cell analysis and display 500,000 461,425 1.3 499,695 0.6
of single samples or in batch format through histogram 750,000 699,500 0.9 744,963 2.9
overlays. The plot on the top is an overlay of histograms
from each of the 7 steps in the dilution series. On the 1,000,000 903,250 0.2 1,003,200 1.4
bottom, 4 replicates at each of two concentrations have 1,500,000 1,241,250 2.5 1,413,195 0.3
been overlaid to demonstrate measurement reproducibility.
Table 1.
Comparative Analysis of Yeast Counts
parallel on the Coulter Counter (Figure 4). While both
instruments demonstrated high linearity in sample
counting (Scepter R2 0.995; Coulter Counter R2 0.998),
Scepter Accuracy in Size Determination
each device lost accuracy at higher concentrations. For 8
Measured Cell Diameter (m)
EMD Millipore, the M mark, and Scepter are trademarks of Merck KGaA, Darmstadt, Germany.
Coulter Counter is a registered trademark of Beckman Coulter, Inc.
Fisher Lit No BN1220113 Lit No. AN3313EN00 Rev. B LS-SBU-11-05576 Printed in the USA. 2/2012
2012 EMD Millipore Corporation, Billerica, MA, USA.