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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Optimization of protocol for immunofluorescence stain to observe nerve infiltration and regeneration in cancer tissue

Hanell, Malin January 2022 (has links)
Background: Neuronal plasticity and regeneration in cancer are understudied aspects of cancer research. Studies have shown that neurogenesis and axonogenesis are associated with cancer progression and metastatic potential. Purpose: The purpose of this project was to optimize an immunofluorescence stain to observe nerve development and regeneration in cancer tissue, with the use of antibodies against neurofilament light chain (Nf-L), growth associated protein 43 (gap-43), and doublecortin X (DCX). Material and method: Staining optimization included evaluations of antigen retrieval, tissue permeabilization, antibody dilution, and duration of primary antibody incubation. The analyses were tested on colorectal- and lung cancer tissues. Results: The detection of Nf-L was not successful in any combination of the analyses or on ether tissue. The staining Gap-43 showed the best results using antigen retrieval with pepsin in HCl and primary antibody dilution 1:500 combined with incubation overnight at 4 °C. Staining for DCX needs more evaluation due to non-specific binding in lung cancer tissue. The stain showed the best results with antigen retrieval performed with pepsin in HCl, primary antibody dilution 1:250 combined with 1 hour incubation at room temperature of the primary antibody. Permeabilization has to some degree shown good results in combination with antigen retrieval with pepsin in HCl. Conclusion: A good protocol was established for Gap-43 detection, but the procedures for Nf-L and DCX detections need to be optimized.

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