Salient Characteristics - MACSQuant Tyto - July 2024 updated 07_24_24.docx

Salient Characteristics of cell sorting equipment Cell sorting is the process through which a particular cell type is separated from other cell populations contained in a sample based on its physical or biological properties, such as size, morphological parameters, viability and both surface and intracellular protein expression. Fluorescence-activated cell sorting (FACS) is the most commonly applied sorting methodology, and the majority of available instruments use high pressure (~70 psi) and electrostatic deflection to select cells in liquid droplets. This approach has two major disadvantages: (1) high pressure can compromise cell viability and functionality; and (2) droplet formation during cell sorting may create biohazard aerosols. Recently, a more advanced technology, based on a low-pressure (<3 psi) and sterile approach using microchip and mechanical valve, has been developed. This new technology allows: (1) gentle separation of cells, which increases cell viability and functionality, without inducing undesirable cell activation during the process; (2) working under sterile conditions during cell separation, which permits every post-sorting use of the selected cell population; (3) performing the approach under a safe environment, since no aerosols are created during cell sorting; and (4) minimal personnel training to perform and use the approach. This technology is revolutionizing the field of cell sorting with impact on immunology, microbiology, and biotechnology, among other scientific areas. At the best of our knowledge, Miltenyi Biotec is the sole source of cell sorter equipment based on a low-pressure and sterile approach using microchip and mechanical valve technology (Product name: MACSQuant® Tyto® Cell Sorter).

Detailed requirements:

1. Cell sorting machine must have 3 lasers (405 nm, 488 nm, and 638 nm) and emission detectors that allow for up to 10 parameters fluorescence-based cell sorting.

2. System must sort cells under sterile low pressure (<3 psi) conditions to maintain maximum cell viability and functionality, producing purified specific cell populations ready for downstream applications.

3. System must have automatic cartridge alignment routine and be operator free sorting process with no laser alignment needed.

4. System must operate with microchip-controlled high-frequency fluidic valve.

5. System must be capable of maintaining cell preparation in the temperature range of 4 to 25°C during cell sorting to maintain cell viability and functionality.

6. System must be able to sort rare immune cell populations from peripheral blood, such as dendritic cells, activated T lymphocytes and B cells, and natural killer cells, among other cells.

7. System must be able to sort sensitive cells from peripheral blood, such as neutrophils.

8. System must include software that allows data acquisition, viewing, and analysis of generated data.

9. System must allow cell sorting under sterile conditions, eliminating the risk of sample-to-sample carry-over or contamination of fluidics.

10. System must have automatic features for flow control, laser alignment, speed detection, and valve timing.

11. System must be benchtop designed and adequate for laboratory environment and space (dimensions should be in the range of 965 mm x 585 mm x 760 mm (W x D x H)).

12. System must be friendly and intuitive to use by lab professionals without intensive training.

13. System must allow working under laboratory biosafety conditions.