NucleoCassette™ / specialized device for viability and cell count Chemometec PCR

Description

The NucleoCassette™ is a specialized device for viability and cell count in the NC-100™. The NucleoCassette™ contains immobilized propidium iodide. When the cassette has been loaded with approximately 60 µl of cell sample mixture the PI is dissolved and mixed with the sample, staining the nuclei of dead cells.
10 pcs / 1 pack . LOT 0121-01 * P/N: 941-0001
Exp. APR 2022
The NucleoCounter® YC-100 system is comprised of the NucleoCounter® YC-100 instrument, NucleoCassette™ and Reagent Y100. An optional but recommended part of the system is the NucleoView™ software. The NucleoCounter® YC-100 is developed as a stand-alone instrument, but it is recommended that the NucleoCounter® YC-100 is connected to a computer using the NucleoView software, which offers a variety of features such as storage of the results and export and printing of the data. Application This protocol for the NucleoCounter® YC-100 system enables the user to obtain absolute volumetric cell count (total cell count) of yeast and thereby to estimate the total number of cells. The protocol is suitable for use on yeast samples, which has low degree of flocculation. It is recommended that NucleoView software is used for this application. Introduction Biomass and biological product produced during a fermentation process are dependent on both cell number and physiological conditions. Whereas the physiological conditions can be controlled during the process, the cell number is mainly controlled by adjusting the initial cell number at the start of the process. Monitoring the cell number allows the scientist to activate inducible systems or to harvest at optimal times for the best product yield. Processes where transformation of yeast is used, can also benefit from this protocol. Traditionally a measurement of a solutions optical density at 600nm (OD600) provides a rough estimate of the number of yeast cells in suspension, but in general a dry cellular weight (DCW) measurement or volumetric cell count provide the best fundament for consistency and optimization of the culturing process. NucleoCounter® YC-100 system provides a fast and accurate DNA based volumetric cell count within seconds, independentl of debris or substrate aggregates in the media. The NucleoCounter® YC-100 is maintenance and calibration free, offering extreme ease of use and reliable operation, and is especially well suited for routine analysis of yeast samples. It is recommended that users validate the NucleoCounter® YC-100 method individually for different yeast strains against their own preferred method of reference for estimating the total concentration of cells or cellular mass, e.g. dry cellular weight, counting chamber with manual estimation of cell concentration, flow cytometry, impedance cell counter, or optical density (OD). Principle In order to determine the total concentration of yeast cells, the cells are treated with Reagent Y100. Reagent Y100 permeates the yeast cell wall and membranes, thereby allowing the cellular DNA to be stained with propidium iodide, which is safely immobilized on the inside of the NucleoCassette. Reagent Y100 has the ability to dissociate minor cell clusters of yeast cells, but not to dissolve or dissociate cells that are in the process of reproductive division.
Approximate correlation between OD, DCW and yeast cell count: 1 OD600 (1 unit optical density measured at 600nm, within linear range) = approx. 0.3 to 0.6 g dry cellular mass/Liter (dry cellular weight, DCW) = approx. 1×106 to 5×106 cells/mL After Reagent Y100 treatment (generally less than 30 seconds), approximately 50 µl of the cell suspension is drawn into the NucleoCassette that contains sufficient amounts of propidium iodide for the staining of the cellular DNA. The NucleoCassette is placed in the NucleoCounter® YC-100 where the cells are automatically enumerated within 30 seconds yielding an absolute volumetric cell count. Using the NucleoCounter® YC-100 system it is possible to determine the total number of yeast cells. Correlation between OD or CDW and volumetric cell counts: A correlation curve may be constructed specifically for each type (and strain) of yeast being measured. Thereby it is possible to calculate the approximate corresponding optical density (OD) or approximate dry cellular weight (DCW) from the cell number measurement using the NucleoCounter® YC-100.
https://chemometec.com/wp-content/uploads/2015/03/994-0300-Total-cell-count-of-yeast-Saccharomyces-cerevisiae-using-the-NucleoCounter-YC-100-system.pdf
https://chemometec.com/wp-content/uploads/2015/03/994-0302-Total-cell-count-and-viability-of-yeast-in-beer-samples-using-the-NucleoCounter-YC-100-System.pdf