1. NucleoCounter YC-100 Presentation

2. 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.

3. 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.

4. Base Technology of ChemoMetec: Fluorescence Microscopy
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.
Emission
CCD
Filter
Lenses
Lenses: Magnification 1.4 X.
Excitation filter: Sorts out green light.
Emission filter: Sorts out long wave length including red light.
Fixture
LED’s
Excitation
Filter

5. Propidium Iodide 1 dye molecule per 4-5 bp
Genome 2.5 x 109 – 4.0 x 109 bp.
C27H34N42+.2I- Mr 668.4
DNA base pair (bp):
A: Adenine
T: Thymine
G: Guanine
C: Cytosine

6. 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

7. 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

8. A Specific Volume is Analyzed

9. 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

10. Total Cell Count Viable Non-viable Lysis buffer (Reagents Y100) PI
Free nucleus PI
PI-stained nucleus
PI-stained nucleus
Viable Non-viable
Lysis buffer
(Reagents Y100)

11. Viability Determination
Viable Non-viable
Viable Non-viable
Free nucleus PI
PI-stained nucleus
PI-stained nucleus
Lysis buffer
(Reagents Y100)
Non-viable cell count
Total cell count

12. Easy Operation

13. NucleoView™ Software

14. The NucleoCassette

15. 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

16. 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

17. 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

18. 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