CELLULAR ACTIVITY QUANTIFICATION USING LABELED PROBES

摘要

Methods and systems for quantifying cellular activity using labeled probes, e.g., quantum dots, are disclosed. In one example approach, a method for quantifying cellular activity in a sample containing intact cells having labeled complexes comprises receiving images of the sample at a plurality of depths and detecting individual intact cells in the images of the sample at the plurality of depths. For each detected cell, discrete labels may be detected and localized in the cell at each depth, a total number of detected and localized labels may be calculated in the cell, and an activity level of the target molecule for the labeled probe in the cell determined.

主权项

1. A computer-implemented method of quantifying the activity level of a target biomolecule in a sample comprising one or more intact cells, said sample having been treated with a reagent, said reagent comprising a label that can label a cellular structure such that the label can be localized within the cell, and a binding component that binds the target biomolecule, said target biomolecule comprising a first activity level within the cell and a second activity level within the sample:
receiving a set of images of the sample, wherein the images within the set comprise individual cells and are taken at a plurality of depths; detecting a first cell in the images of the sample at the plurality of depths; detecting and localizing the label at individual sites in the first cell at each depth in the plurality of depths; calculating a total number of detected and localized labels within the first cell; and calculating a first activity level of the target biomolecule within the cell based on the total number of detected and localized labels at individual sites in the cell; calculating the first activity level of the target biomolecule within a plurality of cells in the sample; and calculating the activity level of the target biomolecule within the sample based on the number of detected and localized labels in the plurality of cells.