Single cell gel electrophoresis
Assignee4D Lifetec AG
InventorsOliver Schicht, Arne-Christian Faisst
Life of this patent
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Abstract
A carrier plate and a liquid sample are provided for the use of a single cell gel electrophoresis device for screening a subject for at least a DNA-damage, in particular a specific DNA-damage, in relation to a predisposition for and/or the presence of a disease. The liquid sample has cells of the subject. A single cell gel electrophoresis on the sample of the subject is performed by positioning the sample accommodated on the carrier plate in the generated homogenous electrical field and controlling strength and direction of the electrical field while the sample is positioned in the gel electrophoresis device. The DNA-damage is determined with a high reproducibility, with a coefficient of variation (CV) of 15% or less, in particular 10% or less, in particular 5% or less.
Claims16
A method for screening cells of a subject for DNA-damage comprising the steps of:
- depositing a liquid sample comprising cells of the subject in at least one spot on a carrier plate, wherein the at least one spot contains a volume of 15 μL or less of the liquid sample, and wherein the at least one spot includes 75 to 400 cells of the subject to be evaluated for DNA-damage;
- performing single cell gel electrophoresis on the at least one spot deposited on the carrier plate;
- and
- determining DNA-damage for the cells of the subject present in the at least one spot based on an analysis of comets appearing in a single image taken of the at least one spot after single cell gel electrophoresis.
A method for screening cells of a subject for DNA-damage comprising the steps of:
- performing single cell gel electrophoresis on at least one spot of a liquid sample comprising cells of the subject deposited on a first carrier plate and on at least one spot of the liquid sample comprising cells of the subject deposited on a second carrier plate, wherein each of the spots contains a volume of 15 μL or less of the liquid sample, wherein each of the spots includes 75 to 400 cells of the subject to be evaluated for DNA-damage;
- and
- determining DNA-damage for the cells of the subject present in the spots based on an analysis of comets appearing in single images taken of the spots after single cell gel electrophoresis;
- wherein a coefficient of variation for the determination of DNA-damage, carrier plate-to-carrier plate, is 15% or less.
A method for determining biological variance between a first subject and a second subject, the method comprising:
- performing single cell gel electrophoresis on at least one first spot a liquid sample comprising cells of the first subject deposited on a carrier plate and on at least one second spot of a liquid sample comprising cells of the second subject deposited on a carrier plate that is the same as or different than the carrier plate on which the liquid sample comprising cells of the first subject is deposited, wherein the first spot contains a volume of 15 μL or less of the liquid sample comprising cells of the first subject, wherein the second spot contains a volume of 15 μL or less of the liquid sample comprising cells of the second subject, wherein the first spot includes 75 to 400 cells of the first subject, and wherein the second spot includes 75 to 400 cells of the second subject;
- determining DNA-damage for the cells of the first subject present in the at least one first spot based on an analysis of comets appearing in a single image taken of the at least one first spot after single cell gel electrophoresis, and determining DNA-damage for the cells of the second subject present in the at least one second spot based on an analysis of comets appearing in a single image taken of the at least one second spot after single cell gel electrophoresis;
- and
- determining the biological variance between the first subject and the second subject based on the determined DNA-damage for the cells of the first subject and the second subject.
Dependencies are inferred from the claim text.