CHROMAVISION aims to develop a pioneering chromosome imaging and manipulation platform to fuel future structural chromosome research. Chromosomal abnormalities are characteristic of many disorders such as cancer, impaired fertility, and neurological disorders. This platform will allow molecular biologists to isolate individual chromosomes from small tissue or cells and deliver them to a super-resolution microscope. Single chromosomes can be brought into focus of the Super-Resolution Correlative Tweezers Fluorescence Microscope (CTFM-SR3D) that is developed in CHROMAVISION. This instrument will for the first time enable 3D, super-resolution, real-time metaphase chromosome observation and manipulation studies under near-physiological conditions. Better imaging and understanding of the chromosomal mechanisms will contribute to our knowledge of the aetiology of human diseases and aid drug discovery. The overall objectives of the proposal are:
1.To develop an integrated and intuitive CTFM-SR3D for chromosome imaging and manipulation that integrates lab-on-a-chip (LOC) microfluidics, multiple-trap optical tweezers and super-resolution fluorescence microscopy
2.To develop and validate production ready prototypes of chromosome labs-on-a-chip for extraction, visualization and actuation of metaphase chromosomes from single cells
3.To develop methods for the quantitative study, in real time, of the 3D structure and dynamics of whole mitotic chromosomes extracted from healthy and diseased single cells
4.To apply the chromosome imaging and manipulation platform to address key questions in biomedical and fundamental chromosome research
Conclusion: Objectives 1, 3 & 4 have been achieved nearly as planned, while objective 2 had to be somewhat modified because chromosome extraction in a lab-on-a-chip setting proved to be very difficult. However, by applying our mitigation plan of extracting chromosomes using more standard bench-top methods and focusing our attention to the development of all-plastic flow chambers, we both managed to avoid any major delays/deviations from the original plans and in fact came, from a commercial point of view, to a better workflow of studying whole metaphase chromosomes. Our general conclusion is that the project was highly successful and has opened new horizons for chromosome research for the coming decade.
