)

Nozzle and pressure selection

The FACSAria II cell separator has three different nozzle orifice sizes (70, 85 and 100 microns) and can separate (sort) cells at different pressures.

The size of the nozzle and the pressure to be used are directly related to the size of the cells to be separated. The larger the cell, the larger the nozzle orifice. When less pressure is used, the speed of separation is slower.

There are general rules for determining the right nozzle size and pressure. However, variations can arise depending on the strength or fragility of the cells to be sorted. This is why it is very important to inform the Flow Cytometry Service if problems arise after separation.

  • 70 microns, high-pressure separation (70 psi):

    • Cells of primary tissues: thymus, spleen, bone marrow

    • Peripheral blood cells

    • Cell lines derived from B and T cells

  • 85 microns, moderate-pressure separation (45 psi):

    • Those listed above that have been transfected/transduced with expression vectors

    • Cells of primary cultures

    • Activated cells

  • 100 microns, low-pressure separation (20 psi)

    • Most cell lines

    •  Adherent cell lines

    • Large, delicate cells of primary tissues (neurons, dendritic cells)

    • Recently transfected cells that express GFP

  • Cell concentration*

    • High pressure: 8-15 million/ml (max. 22,000 cells/sec)

    • Moderate pressure: 7-10 million/ml (max. 12,500 cells/sec)

    • Low pressure: 5-9 million/ml (max. 6,500 cells/sec)

    * These values correspond to the final concentration after the Flow Cytometry Service filters the sample, so the sample should be provided at twice the concentration.

Contactar

Goretti Azparren
Secretaria
Citometría de flujo

General contact:
Avenida Pío XII, 55
31008 Pamplona
España

+34 948 194 700 Ext. 81 1009
mgazparren@unav.es

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