gram ray – getränke qualität
Post on 11-Mar-2022
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Methode
Speed = Security The growth of bacteria, yeast and mold nega-
tively impacts the sensory properties of bever-
ages and influences the safety of the product
for the consumer. An accurate and fast deter-
mination of microorganisms gives the water,
beer, wine or juice producer the long-term se-
curity they need to guarantee the highest qual-
ity at all times.
Targeted development GRAMRAY has been designed to provide fast,
cost-effective and reliable information on all
spoilage microorganisms in beverage produc-
tion. The early warning system reports the
number and species of all living microbes, ide-
ally before problems can arise. Release deci-
sions for produced batches can be made within
the same day instead of 2-7 days.
methods used for antimicrobial susceptibility
testing
Streamline beverage quality
control with confidence GRAMRAY can be integrated into the company's
HACCP system to identify and control microbial
contamination of raw ingredients, in-process
beverages, and finished goods more quickly
with less hands-on labor than with conven-
tional microbiology methods. Finished goods
release decisions are accelerated and compli-
ance with customer specifications can be easily
demonstrated. The system provides the neces-
sary information for the implementation of
quality standards such as IFS 6.1 quickly and
without additional effort.
Sample Prep in 10 Minutes
Simple and fast sample prep
without culturing or amplifica-
tion steps. GRAMRAY can be elegantly integrated into the
traditional beverage sample prep processes.
Low-cost sample substrate 1X, 4 X, 12X up to 96 X sample carriers are avail-
able. The high-quality smooth metal surface
guarantees a high recovery rate and optimal
spectral quality.
Automated sample preparation mibic is developing a fully automated sample
prep solution to enable processing of up to In
288 GRAMRAY samples per day with minimal
hands on time.
Fully automatic measurement All microbes on each of the up to 96 points of a
well plate are counted and identified. The re-
sults are summarized and displayed in a way
that is easy to understand.
Step 1: Concentration Like traditional methods, fil-
tration takes place on a 47
mm 0.45µm membrane.
Step 2: Matrix removal After concentration, the sam-
ple is resuspended, washed
and centrifuged.
Step 3: Dispensing The concentrated and
washed sample is applied to
an inexpensive specialized
sample substrate.
Easy to Use
All information at a glance GRAMRAY quantifies the number of viable mi-
crobial cells and provides phenotypic charac-
terization with species-level classification,
making validation easy by direct comparison to
CFUs recovered by classical microbiology tech-
niques.
Unparalleled sensitivity &
accuracy The combination of single cell analysis and the
high recovery rate (depending on the sample
matrix) makes the sensitivity of the method
better than that of classical methods. GRAMRAY
is independent of the cultivation of bacteria
and is therefore superior to classical methods
in the ability to detect viable but non-cultura-
ble cells. The high accuracy (specificity) of spe-
cies classification is ensured by integrated and
standardized modelling and machine-based
decisions (artificial intelligence).
Simplest display of results The easy-to-use intuitive software turns any-
one into a successful microbial detective after
just 3 days of training. The measurement result
displays each species present and number of vi-
able cells of each. A quality number indicates
the degree of agreement with the species iden-
tification model, allowing for simple assess-
ment of the classification results.
Open microbiological concept In contrast to molecular biological methods,
the GRAMRAY system is open and can be
adapted to meet the needs of specific indus-
tries, processes and products. "House flora"
can be easily integrated into the species classi-
fication model by automated machine learning
and are reliably assigned in future and past
measurements.
Sensitivity + Specificity
Performance Validation To validate the sensitivity of GRAMRAY studies
were done to test recovery rates of yeasts from
red wine samples. Accuracy of species identifi-
cation was tested with a panel of common bev-
erage spoilage yeasts.
Recovery rate > 95%1* • Preparation eliminates disturbing wine ma-
trix residues on > 80% of all cells
• Removed > 99% of all Debris particles
• Resolved cellular aggregates into measura-
ble single cells
Detection limit - 1 cell • Samples were each inoculated with a nom-
inal 2-3 cells of Saccharomyces cerevisiae.
• GRAMRAY has on average 2.5 cells found
per sample
• Perfect match with results from plate culti-
vation
1 The recovery rate reflects the ratio between a measured value and a reference value. The refer-ence value is the number of colony-forming units by classical cultivation on agar plates.
Species Identification Accuracy Specificity > 88% (true positive)
Selectivity > 97% (true negative) • Yeast cells were isolated from white wine,
red wine and water samples using
GRAMRAY.
• 9 genera, 13 species, 19 strains of yeasts
• Cross-validation of the database: sensitiv-
ity and specificity > 99.6%2.
• Strain identification within a species is also
possible.
• GRAMRAY measurement is not subject to
any preconditions - all microbes present in
the model are identified.
2 This value reflects the correctly identified data-base entries from PASTEURX build in model cross validation
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Viable Cell Counts - GRAMRAY
Single Cell Analysis on GRAMRAY • GRAMRAY scans the entire sample surface
and provides an overview image.
• Target images of the single microbes spec-
tral recording allow a precise examination.
• The Raman spectrum is automatically com-
pared with the stored data model.
• The quality of the spectrum and the
correspondence with the data model is
evaluated.
• Number and species of microorgan-
isms is displayed on results.
GRAMRAY – System overview
Control and software package -
LISTER • Simple recipe creation and execution of
measurements
• Optimized high-throughput sample analy-
sis
• User hierarcy-levels
• Highest start-up accuracy and speed in im-
age analysis and Raman spectroscopy
through automated exposure, illumination
modes, autofocus and binarization control
Identification and artificial intel-
ligence - PASTEURX • Software package for modelling for easy in-
tegration of new microorganisms
• Integrated automated spectra comparison
and result by chemotaxono-metric deci-
sion algorithms
Microbial counting and detec-
tion • Measuring interval for image recognition
of individual microbes: 0.5 µm - 50 µm
• Identification for Gram +/- bacteria, yeasts,
fungi and mycobacteria
• Image Analysis for Dark Field Illumination /
Fluorescence
Spectroscopy • Raman system (≤ 20 mW) - optimized for
single cell identification
• Duration per microbe: 1-100 s
• Laser power - selectable from 5% to 100%.
• Spectral resolution - 5 cm-1.
• Wavelength range 200-3.200 cm-1
• Laser safety - Class 1.
• Lens - 100x, 0.9 NA, 1.0 mm WD.
Documentation and results • Original and binarized image data, Raman
spectrum, database match with quality
number
• Number of Viable Cell Counts
Image analysis and sample unit • 2 MP camera for target evaluation and doc-
umentation
• Feedback-controlled motorized x,y,z table
with a restart accuracy of < 100 nm
• Lighting - dark field and fluorescence
• Magnification - 200x, 500x 1000x.
Calibration and self-tests • Integrated for autocalibration and self-
tests
Environmental parameters • Mains voltage + power - 110-230 V, 500 W
• Weight - 52 kg
• Dimensions - 67x58x35 cm
• Temperature range 17-29 ºC,
• Humidity range 10-75 % non-condensing at
29°C
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