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NucleoCounter YC-100 Presentation

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The Value of Objective Yeast Count & Viability• Same Beer with same taste, same color, same foam, same odors…..

• To ensure that the same quality is achieved every time:– To leave out statistical variance.– To leave out personal influence. – To leave out interpretation.

• To trust your method and your final product.

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Yeast Management• High No. of Dead Cells = Poor Yeast Management

– Lower Foam Stability– Higher Turbidity– Lower Flavor Stability– Higher work load for filter

• Correct Pitching of Yeast is Important:– Typical Viability Acceptance Criteria is above 90%– Non Viable Yeast Cells:

• Will act as substrate for bacteria = Infection.• Will decompose = Filter Load, Turbidity, unwanted taste.

– Viable Yeast Cells:• Too many = waste of resources + variation in production time.• Too few = loss of production time + problems with non viable cells.

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Base Technology of ChemoMetec: Fluorescence Microscopy

Fixture

• Lenses: Magnification 1.4 X.

• Excitation filter: Sorts out green light.

• Emission filter: Sorts out long wave length including red light.

Principle: The NucleoCounter is an integrated fluorescence microscope designed to detect signals from the fluorescent dye, propidium iodide (PI) bound to DNA. Results from the NucleoCounter represent either total or nonviable cell concentration, depending on the sample preparation.

LensesCCDEmission

Filter

LED’s

Excitation Filter

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Propidium Iodide

1 dye molecule per 4-5 bp

Genome 2.5 x 109 – 4.0 x 109 bp.

DNA base pair (bp):

A: Adenine

T: Thymine

G: Guanine

C: Cytosine

C27H34N42+.2I- Mr 668.4

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NucleoCounter® YC-100™ • No calibration is a basic advantage of this technique:

– The PI method looks at the DNA of the cell nuclei rather than the whole cells (or particles of a certain size)

– Cell sizes differ, whereas cell nuclei size and amount of DNA or number of base pairs are more uniform

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Calibration Free• No calibration to cell type (size)

• No calibration to volume, as every single analysis volume is exactly known since: The thickness of the measurement chamber of each NucleoCassette is measured during production and coded with black dots on each Cassette. The NucleoCounter reads the code accurately determining the analyzed volume in each measurement.

• Durable optical components makes the NucleoCounter calibration free through out its lifetime

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A Specific Volume is Analyzed

W

T (

co

de

)

Dots for thickness code (T)

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No Cleaning or Maintenance/Service

• The NucleoCassette contains the entire flow system (pump, tubing, mixer, reagents and waste container) as well as the measurement chamber. As the NucleoCassette is discarded after use this means neither cleaning nor maintenance of the NucleoCounter is needed.

• No maintenance as the NucleoCounter is made of very durable and stable components. The NC optics, mechanics and electronics has negligible thermal and aging drift properties. The light source and the linear actuator have been tested to last for >1 x 106 samples.

• No service is needed as no calibration, cleaning or replacement of components is necessary

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Total Cell Count

Viable Non-viable

Viable Non-viable

Free nucleus

PI

PI-stained nucleus

PI-stained nucleus

Viable Non-viable

PI

Lysis buffer

(Reagents Y100)

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Viability Determination

Viable Non-viable

Viable Non-viable

Free nucleus

PI

PI-stained nucleus PI-stained

nucleus

Viable Non-viable

PI

Lysis buffer

(Reagents Y100)

Non-viable cell count Total cell count

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Easy Operation

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NucleoView™ Software

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The NucleoCassette

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The NucleoCassette• Sensitive parts, moving parts and “wear and tear” parts have been taken out of the

instrument and integrated in the disposable NucleoCassette

•Pump•Tubing’s•Mixer•Precision optics•Chemicals•Calibration•Waste container

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Reagent Y100• The lysis reagent Y100 disrupts the yeast cell walls and membranes, rendering the

nuclei susceptible to staining with propidium iodide (PI).

• Dissolves small cell aggregates

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NucleoCounter® YC-100™ Key Advantages• Easy operation – no preparation• To be used in Lab & Production (24 h)• To be used by untrained operator• 30 sec. analysis time • Calibration free• No cleaning• Maintenance and service free• Space saving• Safe sample handling & disposal • High precision & repeatability• Good reproducibility

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NucleoCounter® YC-100™ SUMMING UP• Gives the head office control of yeast management• Insures the same reference standard at all locations• The NucleoCassette insures calibrated volume with every measurement• Can be used in lab and production (24h) and is not dependent of a computer – can

be used as a stand alone unit• Easy to implement at all locations• Extreme ease of use – very limited training is required• Well proven and preferred technology for automatic counting in breweries world

wide• Robust technology for all environments• Handles clustering cells extremely well