Clearing factor

In centrifugation the clearing factor or k factor represents the relative pelleting efficiency of a given centrifuge rotor at maximum rotation speed. It can be used to estimate the time t {\displaystyle t} (in hours) required for sedimentation of a fraction with a known sedimentation coefficient s {\displaystyle s} (in svedbergs):

t = k s {\displaystyle t={\frac {k}{s}}}

The value of the clearing factor depends on the maximum angular velocity ω {\displaystyle \omega } of a centrifuge (in rad/s) and the minimum and maximum radius r {\displaystyle r} of the rotor:

k = ln ( r m a x / r m i n ) ω 2 × 10 13 3600 {\displaystyle k={\frac {\ln(r_{\rm {max}}/r_{\rm {min}})}{\omega ^{2}}}\times {\frac {10^{13}}{3600}}}

As the rotational speed of a centrifuge is usually specified in RPM, the following formula is often used for convenience:[1]

k = 2.53 10 5 × ln ( r m a x / r m i n ) ( R P M / 1000 ) 2 {\displaystyle k={\frac {2.53\cdot 10^{5}\times \ln(r_{\rm {max}}/r_{\rm {min}})}{({\rm {{RPM}/1000)^{2}}}}}}

Centrifuge manufacturers usually specify the minimum, maximum and average radius of a rotor, as well as the k {\displaystyle k} factor of a centrifuge-rotor combination.

For runs with a rotational speed lower than the maximum rotor-speed, the k {\displaystyle k} factor has to be adjusted:

k a d j = k ( maximum rotor-speed actual rotor-speed ) {\displaystyle k_{\rm {adj}}=k\left({\frac {\mbox{maximum rotor-speed}}{\mbox{actual rotor-speed}}}\right)} 2

The K-factor is related to the sedimentation coefficient S {\displaystyle S} by the formula:

T = K S {\displaystyle T={\frac {K}{S}}}

Where T {\displaystyle T} is the time to pellet a certain particle in hours. Since S {\displaystyle S} is a constant for a certain particle, this relationship can be used to interconvert between different rotors.

T 1 K 1 = T 2 K 2 {\displaystyle {\frac {T_{1}}{K_{1}}}={\frac {T_{2}}{K_{2}}}}

Where T 1 {\displaystyle T_{1}} is the time to pellet in one rotor, and K 1 {\displaystyle K_{1}} is the K-factor of that rotor. K 2 {\displaystyle K_{2}} is the K-factor of the other rotor, and T 2 {\displaystyle T_{2}} , the time to pellet in the other rotor, can be calculated. In this manner, one does not need access to the exact rotor cited in a protocol, as long as the K-factor can be calculated. Many online calculators are available to perform the calculations for common rotors.

References

  1. ^ "Basics of Centrifugation"
  • Beckman Coulter lab resources and manuals
  • Appendix F of the Cell Biology Laboratory Manual
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