University of Twente Student Theses
Optimizing Fluorescent Immunostaining Protocols for Retinal Organoids
Blaauw, Meike (2024) Optimizing Fluorescent Immunostaining Protocols for Retinal Organoids.
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| Abstract: | Retinal degenerative diseases (RDs) are a leading cause of visual impairment, requiring advanced models for better understanding and treatment. Retinal organoids (ROs), derived from human induced pluripotent stem cells (hPSCs), have emerged as promising tools due to their ability to mimic human retinal structures more closely than traditional animal models. This study aims to optimize immunofluorescence (IF) staining techniques for ROs to enhance the visualization of cellular structures and improve the accuracy of disease modelling and therapeutic testing. The research focused on two primary methods: IF staining of retinal organoids cryosections and of whole retinal organoids. The whole organoid staining protocols were adjusted for the three-dimensional structure of the ROs, incorporating a clearing step to allow deeper imaging. Detailed imaging using a Zeiss confocal microscope provided comprehensive insights into the effectiveness of different staining conditions. Optimal antibody concentrations were identified, with specific protocols improving the visualization of cell morphology and positions within the retinal tissue. It also highlighted the benefits of a simple immersion clearing technique, which improved imaging depth and the visibility of internal structures by decreasing light scattering. The study highlights the importance of optimized IF staining protocols for advancing retinal organoid research. Refining these techniques aims to improve the understanding of retinal diseases, enabling more precise drug testing and the development of effective therapeutic strategies. These advancements hold significant potential for enhancing the consistency and reproducibility of retinal disease models, ultimately contributing to better patient outcomes. |
| Item Type: | Essay (Bachelor) |
| Faculty: | TNW: Science and Technology |
| Subject: | 42 biology |
| Programme: | Biomedical Technology BSc (56226) |
| Link to this item: | https://purl.utwente.nl/essays/103828 |
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