Process and system for analyzing the expression of biomarkers in a cell

1. A process for displaying cells within a cellular sample comprising:

a. acquiring a digital image of a cell, through microscopic examination, using multiplex sequential staining to identify multiple biomarkers within the cell comprising;

iteratively staining and bleaching a slide containing the cellular sample by;

capturing a first image;

using a feature of the first image as a reference;

capturing one or more successive images such having a same field of view as the first image to create an image stack;

aligning each successive image, in the image stack with the first image using the first image as a reference;

transforming the image stack into a digital image;

b. transforming the digital image into a dataset comprising a plurality of data points, and placing the dataset in a registry, wherein each data point is representative of a level of expression of one of the multiple a biomarkers in the cell and where the level of expression is associated with each pixel from the level of signal from that pixel of the label for that biomarker from the sequential staining;

c. interrogating said dataset for data points having a similar pattern of expression of said biomarkers using an algorithm wherein such similarity is determined by a numerical analysis that uses the level of expression of each of the biomarker as a semi-continuous variable;

d. identifying two or more data points from step c, having minimum variance;

e. grouping the data points having the minimum variance together to create a group of biomarkers whose members have a similar biomarker expression profile and assigning said group a new biomarker expression profile represented by a new data point in a new dataset, wherein said new data point is based on a central value for each attribute considered and the grouping is based on a hierarchical approach assigning the data group to the group with a similarity value closes to the biomarker expression profile;

f. repeating steps c, d, and e until a predetermined number of groups is generated whereby the number of groups is determined using a hierarchical approach; and

g. overlaying an image of of the resulting grouping of step f, wherein the image is registered to the digital image of the cell and creating a new image corresponding to a biomarker expression profile of said cell, wherein the groups individually represent at least one subcellular component comprising a subcellular compartment selected from a nucleus, cytoplasm or membrane.

2. The process of claim 1 wherein the image displays the groups by sequentially overlaying the groups on the digital image and by which each group is given a separate visible designation.

3. The process of claim 2 wherein the visible designation is a color, symbol, or combination thereof.

4. The process of claim 1 wherein the similarity value is based on a Euclidean distance in biomarker space between each data set.

5. The process of claim 1 wherein the similarity value is calculated using classical statistical analysis, a learning algorithm, or a combination thereof.