The methods and reagents described in this invention are used to analyze circulating
tumor cells, clusters, fragments, and debris. Analysis is performed with a number of platforms, including
flow cytometry and the CellSpotter® fluorescent
microscopy imaging
system. Analyzing damaged cells has shown to be important. However, there are two sources of damage:
in vivo and
in vitro. Damage
in vivo occurs by
apoptosis,
necrosis, or immune response. Damage
in vitro occurs during sample acquisition, handling, transport,
processing, or analysis. It is therefore desirable to confine, reduce, eliminate, or at least qualify
in vitro damage to prevent it from interfering in analysis. Described herein are methods to diagnose, monitor, and screen
disease based on circulating rare cells, including
malignancy as determined by CTC, clusters, fragments, and debris. Also provided are kits for assaying biological specimens using these methods.