Sheath flow is a surrounding liquid stream that carries and positions a sample-bearing stream. In a conventional flow cytometer, the sample enters an enclosing flow through the flow cell; hydrodynamic focusing guides the core toward the optical interrogation region. The FACSCelesta technical description identifies the separate pressurized sample input and surrounding sheath fluid.[ref]
BD FACSCelesta Flow Cytometer User’s Guide by BD Biosciences. Chapter 7, About fluidics, Hydrodynamic focusing
The interface between the streams is not a solid tube. In laminar flow, adjacent fluid can travel with relatively little bulk transverse mixing while dissolved material still crosses the interface by diffusion. Positioning particles, controlling sample dilution and preventing molecular exchange are therefore different objectives.
An original diffusion comparison makes the distinction quantitative. A stream travels an observation distance at characteristic speed , giving residence time . For one transverse coordinate, a passive species with diffusion coefficient has characteristic displacementTake stipulated s and a core radius of . A species with has , whereas one with in those units has . The same focusing geometry can confine slowly diffusing particles well while allowing appreciable redistribution of a smaller solute. These coefficients are illustrative, not assigned to a particular reagent or cell.
Increasing sheath relative to sample flow can narrow the core, but the quantitative relation depends on the velocity field and geometry. The hydrodynamic focusing article contrasts equal-speed and Poiseuille flow models. A flow fraction is not automatically an area fraction.
A useful experimental check keeps a stable reference-particle suspension and optical settings fixed while changing the flow balance. Event-rate changes, signal spread and background distinguish throughput from measurement consistency. Clean fluid paths, suitable fluid composition and detector controls address different parts of the experiment; a well-centered sample stream alone does not certify a quantitative fluorescence measurement.
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