Quantitation of cellular DNA and cell numbers using element labeling
View Patent ↗Methods and kits for the quantitation of cellular DNA and cell numbers are provided. Passive element uptake, element-labeled DNA intercalators, and element labeled affinity reagents are used to quantify DNA and cells. The DNA and the cells are analyzed by elemental analysis, including ICP-MS. The methods and kits provide a fast and accurate analysis of cellular DNA and cell numbers.
1. A method to determine DNA content and antigen content per cell in a sample of cells, comprising:
(a) fixing and permeabilizing of the cells;
(b) incubating the sample of cells with an intercalator tagged with a first element under conditions appropriate to incorporate the intercalator into the DNA within the cells of the sample;
(c) removing unbound intercalator from the cells;
(d) incubating antigen specific affinity products labeled with a second element tag with the cells;
(e) removing unbound affinity products from the cells;
(f) simultaneously measuring the intercalator tagged with a first element and the affinity product labeled with the second element tag by elemental analysis using an Inductively Coupled Plasma Mass Spectrometer; and
(g) calculating the DNA content and antigen content per cell.
2. The method of claim 1 where in (d) the antigen specific affinity product is an antibody complex comprising a secondary antibody labeled with an element tag that binds to a primary antibody, and wherein the primary antibody is specific to the antigen.
3. The method of claim 1 wherein (d) the cells are incubated with at least two classes of antibodies, each class of antibodies specific to different classes of antigens and each class of antibody is labeled with unique element tag.
4. The method of claim 1 further comprising a step after step (d) wherein the cells are dissolved to generate a relatively homogeneous sample before determining the amount of metal by performing solution elemental analysis.
5. The method of claim 4 wherein the cells are dissolved in an acid solution.