Examples of projects completed

We have completed numerous projects for researchers across various fields of study. Our advanced equipment and skilled staff have enabled us to provide high-quality imaging and analysis services for a range of applications. Here are a few examples of the projects we have completed:

Developing a tissue-specific quantification method of ribosome activity in intact C. elegans using fluorescence microscopy and image analysis: Caloric restriction is known to be a pro-longevity treatment in C. elegans thought the decrease in protein translation. Majority of the studies analyze translation on lysates from pool of worms, however, growing evidence suggest that difference in translation at the tissue level may exist and play an important role in aging. On this project, I helped to develop and optimized the imaging set-up as well as the image analysis workflow. By using OP-puromycin and fluorescence microscopy, we were able to quantify relative protein translation rates across tissues in intact C. elegans. This work has been done in collaboration with Dr. Rollins at the MDI Biological Laboratory. This project has been publish in Cell Reports Methods:

Somers H.M., Fuqua J.H., Bonnet F.X.A., Rollins J.A. Quantification of tissue-specific protein translation in whole C. elegans using O-propargyl-puromycin labeling and fluorescence microscopy, Cell Reports Methods, Volume 2, Issue 4, 2022, 100203

Graphical abstract of the paper

Imaging and data analysis of biomolecular condensates, such as C. elegans germ granules: Germ granules are conserved ribonucleoprotein condensates found in the germline of almost all animals. Here they retain a memory of germline-specific expression through cell divisions and across generations. Their unique attributes require innovative high-resolution imaging techniques and quantitative measures, which I have helped to develop in collaboration with Dr. Updike at the MDI Biological Laboratory. For example, one C. elegans germ-granule component called LOTR-1 demixes into a specific sub-granules called Z granules. I helped develop imaging techniques and quantitative measures to gauge the extent of LOTR-1 demixing, providing insight into the function of its mammalian homolog TDRD5 during spermatogenesis. This project has been publish in PLOS Genetics:

E.A. Marnik, M.V. Almeida, P.P Cipriani, G. Chung, E. Caspani, E. Karaulanov, H.H. Gan, J. Zinno, I.J. Isolehto, F. Kielisch, F. Butter, C.S. Sharp, R.M. Flanagan, F.X. Bonnet, F. Piano, R.F. Ketting, K.C. Gunsalus, D.L. Updike (2022) The Caenorhabditis elegans TDRD5/7-like protein, LOTR-1, interacts with the helicase ZNFX-1 to balance epigenetic signalshttps://journals.plos.org/plosgenetics/article?id=10.1371/journal.pgen.1010245 in the germline. PLoS Genet 18(6): e1010245. https://doi.org/10.1371/journal.pgen.1010245

D) Live cell imaging of GFP::LOTR-1 and PGL-1::RFP in the germline of living worms using a wide field microscope . E) Super-resolution confocal imaging using the Zeiss 980 with airy scan 2 of GFP::LOTR-1 with PGL-1::RFP (top) and GFP::LOTR-1 with RFP::ZNFX-1 (bottom) in pachytene germ cells. F) Comparison of the center of mass difference from LOTR-1 for PGL-1 or ZNFX-1.

Personalized maco on FIJI to step up image processing: Documentation and screening are time consuming and laborious tasks when it comes to enhance, rotate, cropped and organized images. To speed up these steps the LMF can write personalized maco for FIJI. The following macro has been written to process automatically several images from the Zeiss stereoscope axiozoom V16 located in a folder.

Screenshot of the macro
Maco in action

These examples illustrate our diverse capabilities and expertise in providing customized microscopy solutions to meet our users’ specific research needs. Contact us to learn how we can help advance your research with our high-quality imaging and analysis services.

Contact fbonnet@mdibl.org today to learn more about our services and how we can help you achieve your research goals. We look forward to working with you and unlocking the mysteries of the microscopic world together!