摘要:
Automated methods and devices that facilitate iterative staining of biological samples from imaging applications are provided. The methods include the steps of providing a small volume flow cell containing a biological sample, applying a stain to the biological sample, combining at least two precursor reagents to form an activated destaining agent and wherein the activated destaining agent decomposition rate is greater than or similar to the destaining reaction rate, flowing the destaining agent over the biological sample at a flow rate that is greater than the decomposition rate of the activated destaining agent, and releasing the sample from the flow cell wherein the integrity of the sample is intact. The process of staining, combining and flowing may be iteratively repeated. Also disclosed herein are devices for iterative staining of biological samples comprising a flow cell, in fluid communication with a premixer, wherein the volume capacity of the premixer is smaller than about five times the volume capacity of the flow cell.
摘要:
Novel methods of probing multiple targets in a biological sample are provide whereby the targets are DNA, RNA and protein. The method comprises subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe that binds an RNA target, observing a signal, and optionally removing the signal. The method further comprises an antigen retrieval protocol, observing a signal, removing the signal, and optionally applying a protease treatment to access the sample's DNA targets by subjecting the sample to an in situ hybridization reaction using a labeled nucleic acid probe, observing a signal from the labeled DNA targets, and optionally removing the signal.
摘要:
The present application discloses a technique for obtaining and storing data on expression of multiple biomarkers in individual cells or the compartments of individual cells in a tissue specimen and methods of utilizing that data to create groups, the members of which share some degree of similarity greater than the general population from which the data is drawn, by an analysis of digital images of a portion of the tissue specimen which has been iteratively stained to generate optical signals, typically fluorescent, reflective of the amount of each of the biomarkers examined. The analysis of the images involves a segmentation routine whereby each pixel of the examined images is assigned to an individual cell or a compartment of an individual cell, the intensity of the signal representative of each biomarker is measured for each pixel, a dataset is created in which each cell or compartments of each cell is associated with a signal intensity for each biomarker examined, and the dataset is interrogated with appropriate numerical tools to create groups. It also discloses the display of such groups on images of the tissue examined such that the individual cells belonging to a particular group are marked or indicated on one of the images examined. It further discloses examining the biomarker data for each group for possible association with a biological condition or process in cases in which tissue specimens drawn from different subjects or different portions of the tissue of a subject have been examined. It also discloses using this examination in the diagnoses, prognoses or assessment of the response to therapy of a condition or disease.