Capturing, Analyzing and Collecting Adherent Cells Using Microarray Technologies
Effective separation of a particular cell ofinterest from a heterogeneous cell population is crucial to many areas of biomedical research including microscopy, clinical diagnostics and stem cell studies. Examples of such studies include the analysis of singlecells, isolation of transfected cells and cell transformation studies. It is important to have a technique capable of identifying the desired cells, separating these cells from unwanted cells and collecting the marked cells for further analysis. Two new technologies, referred to as micropallets and microrafts, will be described for isolating adherent cells with extremely high post-sorting purity and viability.
These devices comprise arrays of microelements weakly attached to a substrate. Following culture of adherentcells on the elements, individual microstructures are selectively detached from the array while still carrying the cells. During my presentation I will describe new approaches for capturing, examining and isolating individual cells by micropallet and microraft technologies. A new approach was developed to isolate released microstructures from the array employing magnetism. Microstructures were embedded with uniformly dispersed magnetic nanoparticles which allowedcollection by an external magnet immediately following release.
Application of a magnetic field permitted microstructure collection with high yield, precision and purity. This improved collection efficiency enabled isolation of very rare cell types. Large arrays constituting over one million micropallets were developed along with imaging analysis software to identify and sort low abundance target cells. This system was employed to isolate pancreatic cancer stem cells from a heterogeneous cell population and circulating tumor cellsdirectly from peripheral blood. Additionally, an array-based cell colony replication strategy was established which allowed highly efficient colony splitting and sampling.