Centrifugal technology has been widely used in the biological sciences, especially in the fields of biochemistry and molecular biology. Each biochemistry and molecular biology laboratory is equipped with a variety of centrifuges. Centrifugal technology is mainly used for the separation and preparation of various biological samples. Under the high-speed rotation of the biological sample suspension, the suspended micro-particles (organelles, precipitation of biological macromolecules, etc.) settle at a certain speed due to the huge centrifugal force. Thereby separating from the solution, the rate of settling depends on the quality, size and density of the particles.
Basic working principle When a particle (biomacromolecule or organelle) is subjected to centrifugal force under high-speed rotation, the centrifugal force "F" is defined by:
F = m•a = m•ω2 r
a — the acceleration of the particle rotation, m — the effective mass of the settled particle, ω—the angular velocity at which the particle rotates, and r—the radius of rotation of the particle (cm).
Usually the centrifugal force is usually expressed as a multiple of the Earth's gravity, hence the relative centrifugal force "RCF". Or by multiplying the number by "g", for example 25000 x g, the relative centrifugal force is 25000. Relative centrifugal force means that in the centrifugal field, the centrifugal force acting on the particles is equivalent to the multiple of the earth's gravity, and the unit is the gravitational acceleration "g".
(980cm/sec2), at this time the "RCF" relative centrifugal force can be calculated by the following formula:
∴ RCF = 1.119×10-5×(rpm) 2 r
( rpm — revolutions per minute revolutions per minute, r/min )
As can be seen from the above equation, as long as the radius of rotation r is given, the RCF and the rpm can be converted to each other. However, due to the difference in shape and structure of the rotor, the distance between each point of the centrifuge tube of each centrifuge from the nozzle to the bottom of the tube is different, so the calculation is to specify the radius of rotation. The average radius "ra v" is substituted for:
Ra v=( r min+rmax) / 2
The measurement of r is shown below:
34o
Rmin
Rotary axis rav
Rmax
In general, low-speed centrifugation is often expressed in terms of "rpm" and high-speed centrifugation is indicated by "g". When calculating the relative centrifugal force of the particles, it should be noted that the distance "r" between the centrifuge tube and the center of the rotating shaft is different, that is, the position of the settled particles in the centrifuge tube is different, and the centrifugal force is also different. Therefore, when reporting ultracentrifugation conditions, it is common to always replace the revolutions per minute "rpm" with a multiple of gravity "xg" because it can truly reflect the centrifugal force of the particles at different locations within the centrifuge tube and its dynamic changes. The data on centrifugal force in the scientific literature usually refers to its average value (RCFa v), which is the centrifugal force at the midpoint of the centrifuge tube.
In order to facilitate the conversion between the rotational speed and the relative centrifugal force, Dole and Cotzias made a nomogram of the relationship between the rotational speed "rpm", the relative centrifugal force "RCF" and the rotational radius "r" using the RCF calculation formula. It is more convenient than the formula calculation method (see the appendix for the column diagram). When converting, first take the known radius on the r scale and take the known number of centrifuge revolutions on the rpm scale, then draw a straight line between the two points, which corresponds to the intersection on the RCF scale in the figure. Relative centrifugal force values. Note that if the known rotation value is on the right side of the rpm scale, the value on the right side of the RCF scale should be read. If the rotation value is on the left side of the rpm scale, the value on the left side of the RCF scale should be read.
Centrifugal technology applied in biology
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