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Centrifugation separates suspended material by its motion relative to a rotating liquid. A centrifuge rotor creates an outward centrifugal acceleration in the rotating reference frame; material denser than its surroundings tends to move outward, opposed by drag force. Collected material can form a centrifugation pellet, leaving a supernatant, or remain in separated zones within a density gradient. The useful question is which physical difference the separation exploits: a difference in sedimentation coefficient or a difference in buoyant density.

Centrifugation makes otherwise slow settling useful for studying biological cells, organelles, proteins and DNA. Separation and identification are different steps: a centrifugation pellet is a collected fraction, not proof that it contains only one kind of organelle. The lysozyme wall-removal experiment illustrates a complementary readout: testing material released into the separated liquid can be more informative than inspecting the retained solids.

A centrifuge rotor holds sample vessels around the rotation axis. A fixed-angle centrifuge rotor holds tubes at a fixed inclination; a swinging-bucket centrifuge rotor lets them swing outward during rotation. Material moves away from the axis, so a centrifugation pellet can first collect against an outer tube wall rather than travel along the entire tube axis. Geometry sets both the rotor radius and the travel distance.^[ref]

Relative centrifugal force is the dimensionless ratio of the local centrifugal acceleration magnitude $a$ to the conventional gravitational reference $g_0=9.80665\mathrm{m}{\mathrm{s}}^{-2}$. Despite its name, it is an acceleration ratio: the actual force on a specified mass $m$ is $ma$. With angular speed $\omega$ and rotor radius $r$, $a={\omega }^{2}r$. If $n$ is the numerical rotational speed in revolutions per minute and $r_{\mathrm{cm}}$ the numerical rotor radius in centimetres, unit conversion givesThus rpm alone does not specify the centrifugal acceleration.^[ref]

For a stipulated $3000\mathrm{rpm}$, rotor radii of $5$ and $10\mathrm{cm}$ give approximately $503$ and $1006$ times $g_0$. Doubling the rotor radius doubles relative centrifugal force; doubling rpm quadruples it. Even equal relative centrifugal forces do not guarantee equal separation: travel distance, elapsed time, viscosity, sample density and particle shape still matter. These are comparisons of a mathematical model, not instructions for running an instrument.

The rotor radius $r$ is the perpendicular distance from the rotation axis to a specified sample position, not the tube's length. A filled tube spans a range $r_{\min }\le r\le r_{\max }$. Consequently, one rotational speed produces different local centrifugal accelerations along that range. A reported relative centrifugal force needs a stated reference position, such as the outermost sample radius.