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Content archived on 2024-04-30

The development of scanning probe microscopy techniques for the study of genetic deletions


Research objectives and content
Scanning probe microscopy (SPM) is a family of microscopy techniques very promising in the study of biological applications. Atomic force microscopy can provide high resolution (<1 nm) topographic images of biological structures in air and liquid environment, preserving the native structure of the biological sample. Optical imaging has played a important role in biological investigations for a very long time. The near-field scanning optical (NSOM) can go beyond the diffraction-limited resolution of conventional optical microscopes to give optical images with resolution of about 20 nm. The objective of the project is the development of these SPM techniques and other recent SPM techniques such as tapping-mode phase contrast imaging or non-contact shear force microscopy (ShFM) to tackle an important problem in medicine of chromosome deletions. These chromosome losses (and gains) are of the fundamental importance in cancer and in birth defects. Although many of these chromosome abnormalities can be detected with conventional optical microscopy ; many cannot. SPM will allow not only the detection but also the pinpointing of the location of such sites. The different SPM techniques will be developed and combined to determine the banding pattern and the volume of the chromosomes. We will also expect to locate specific genes and high transcriptional bands. These studies provide the baseline ( for comparison of chromosomes with genetic deletions. The comparison will begin with mayor deletions and work towards microdeletions, not detectable by conventional cytogenetic methods. Finally, some attemp will be made to 'unwind' chromosome in-situ using a combination of chemical treatment and the AFM tip as a micromechanical tool in order to reveal further information on its structure.
Training content (objective, benefit and expected impact)
The group of the Bristol University is expert in SPM techniques as well as instrumentation development such as NSOM. This | group has recently developed a non-invasive SPM technique, the ShFM. Both techniques are very promising for biological applications and are little known in my country. I would like to learn and develop these SPM techniques and complete my | experience in SPM techniques (STM, AFM). Also, this group together experts of the university work in biological applications. | This will definitely be the case for the chromosome studies where Prof. Soothill and Dr. Thein (University of Bristol/St | Michaels Hospital), experts in genetic defects in chromosomes and in the preparation of appropriate specimens for microscopy, | will collaborate closely in this project. These experts together the biological background of the group will provide a important | training in molecular biology and more specificly in the study of chromosomes and genetic defects.
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