Centrifuge rotor mounted on drive shaft
US-10486170-B2 · Nov 26, 2019 · US
US10436685B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-10436685-B2 |
| Application number | US-201715417858-A |
| Country | US |
| Kind code | B2 |
| Filing date | Jan 27, 2017 |
| Priority date | Jul 30, 2014 |
| Publication date | Oct 8, 2019 |
| Grant date | Oct 8, 2019 |
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A method for processing particles contained in a liquid biological sample is presented. The method uses a rotatable vessel for processing particles contained in a liquid biological sample. The rotatable vessel has a longitudinal axis about which the vessel is rotatable, an upper portion having a top opening for receiving the liquid comprising the particles, a lower portion for holding the liquid while the rotatable vessel is resting, the lower portion having a bottom, and an intermediate portion located between the upper portion and the lower portion, the intermediate portion having a lateral collection chamber for holding the liquid while the rotatable vessel is rotating. The method employs dedicated acceleration and deceleration profiles for sedimentation and re-suspension of the particles of interest.
Opening claim text (preview).
We claim: 1. A method for separating particles from a liquid biological sample and re-suspending the particles in a secondary liquid, the method comprising: a) introducing the liquid biological sample comprising the particles into a rotatable vessel through a top opening of the rotatable vessel such that the liquid is held by a lower portion of the rotatable vessel, wherein the rotatable vessel comprises a longitudinal axis about which the vessel is rotatable, an upper portion comprising the top opening for receiving the liquid comprising the particles, the lower portion for holding the liquid while the rotatable vessel is resting, the lower portion comprising a bottom, and an intermediate portion located between the upper portion and the lower portion, the intermediate portion comprising a lateral collection chamber for holding the liquid while the rotatable vessel is rotating; b) rotating the rotatable vessel about its longitudinal axis at a rotational speed, wherein the liquid comprising the particles is moved to the lateral collection chamber by centrifugal force, and wherein the centrifugal force is sufficient to sediment the particles in a region of sedimentation of an inner wall of the lateral collection chamber; c) decelerating at a rate of from 50 rpm/s to 1000 rpm/s and ultimately stopping the rotation of the rotatable vessel, wherein the liquid flows back to the lower portion of the rotatable vessel, wherein an angular deceleration is not sufficient to detach the particles from the inner wall of the lateral collection chamber by causing shearing forces between wall and liquid such that at least a part of the particles remain attached to the inner wall of the lateral collection chamber, thereby separating at least the part of the particles from the liquid; d) withdrawing the liquid from the bottom of the rotatable vessel while leaving the particles in the lateral collection chamber; e) adding the secondary liquid to the rotatable vessel through its top opening; f) rotating the rotatable vessel about its longitudinal axis in a first direction; and g) decelerating and ultimately stopping the rotation of the rotatable vessel, wherein an angular acceleration in step f) and/or an angular deceleration in step g) is sufficient to detach at least a part of the particles from the inner wall of the lateral collection chamber by causing shearing forces between wall and liquid, thereby re-suspending the particles in the secondary liquid and wherein the angular acceleration in step f) has a rate of at least 500 rpm/s while the angular deceleration in step g) has a rate of at least 500 rpm/s. 2. The method according to claim 1 , wherein steps f) and g) are repeated one or more times. 3. The method according to claim 1 , further comprising after step g), h) rotating the rotatable vessel about its longitudinal axis in a second direction opposite to the first direction at a rotational speed; and i) decelerating and ultimately stopping the rotation of the rotatable vessel, wherein an angular acceleration in step h) and/or an angular deceleration in step i) is sufficient to detach at least a part of the particles from the inner wall of the lateral collection chamber by causing shearing forces between wall and liquid and wherein the angular acceleration in step h) has a rate of at least 500 rpm/s while the angular deceleration in step i) has a rate of at least 500 rpms/s. 4. The method according to claim 3 , wherein steps f) and g), steps h) and i), and/or steps f) through i) are repeated one or more times. 5. The method according to claim 1 , wherein the rate of deceleration in step c) is smaller than any of the rates of acceleration or deceleration in steps f) or g). 6. The method according to claim 1 , wherein the rate of deceleration in step c) is from 50 rpm/s to 400 rpm/s.
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