Publicação
Protocol for semi-automatic quantitative bioimaging analysis of synapse loss
| Resumo: | Synapse loss correlates with cognitive decline in neurodegenerative diseases like late-onset Alzheimer’s disease. We developed a semi-automated workflow to quantify synapse loss in primary mouse neurons. The protocol includes culturing neurons on coverslips, using short hairpin RNA (shRNA)-expressing lentivirus, maintaining cultures, and labeling excitatory and inhibitory presynaptic markers by immunofluorescence. Image acquisition and analysis using Fiji/ComDet macros enable region of interest selection, neurite length measurement, puncta detection, and quantification of synapse density, size, and intensity following Bin1 knockdown. For complete details on the use and execution of this protocol, please refer to Barata et al. |
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| Autores principais: | Barata, Mariana A. |
| Outros Autores: | Guimas Almeida, Cláudia |
| Assunto: | Cell-based Assays Microscopy Neuroscience General Neuroscience General Immunology and Microbiology General Biochemistry,Genetics and Molecular Biology |
| Ano: | 2026 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| Resumo: | Synapse loss correlates with cognitive decline in neurodegenerative diseases like late-onset Alzheimer’s disease. We developed a semi-automated workflow to quantify synapse loss in primary mouse neurons. The protocol includes culturing neurons on coverslips, using short hairpin RNA (shRNA)-expressing lentivirus, maintaining cultures, and labeling excitatory and inhibitory presynaptic markers by immunofluorescence. Image acquisition and analysis using Fiji/ComDet macros enable region of interest selection, neurite length measurement, puncta detection, and quantification of synapse density, size, and intensity following Bin1 knockdown. For complete details on the use and execution of this protocol, please refer to Barata et al. |
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