Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers
Summary: Despite being among the largest neurons in the mammalian brain, Purkinje cells are difficult to visualize and trace via immunofluorescence because their dendritic arbors extend through several cerebellar layers. This protocol describes a two-antibody strategy we developed to study Purkinje...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-06-01
|
| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722003070 |
| _version_ | 1818207476663713792 |
|---|---|
| author | Salvatore Botta Alexei Chemiakine Vincenzo A. Gennarino |
| author_facet | Salvatore Botta Alexei Chemiakine Vincenzo A. Gennarino |
| author_sort | Salvatore Botta |
| collection | DOAJ |
| description | Summary: Despite being among the largest neurons in the mammalian brain, Purkinje cells are difficult to visualize and trace via immunofluorescence because their dendritic arbors extend through several cerebellar layers. This protocol describes a two-antibody strategy we developed to study Purkinje cell morphology in mice. With it, one can reconstruct three-dimensional images of Purkinje cells at single-neuron resolution across multiple layers. The substantially improved image quality reveals subtle defects, enabling more meaningful morphological analysis.For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2015). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-12-12T04:29:32Z |
| format | Article |
| id | doaj.art-b1eda11c47ba4cc99884cd9424ff8fff |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-12T04:29:32Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-b1eda11c47ba4cc99884cd9424ff8fff2022-12-22T00:38:08ZengElsevierSTAR Protocols2666-16672022-06-0132101427Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layersSalvatore Botta0Alexei Chemiakine1Vincenzo A. Gennarino2Department of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USA; Department of Translational Medical Science, University of Campania Luigi Vanvitelli, 80131 Napoli, ItalyDepartment of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USADepartment of Genetics and Development, Columbia University Irving Medical Center, New York, NY 10032, USA; Departments of Pediatrics and Neurology, Columbia University Irving Medical Center, New York, NY 10032, USA; Columbia Stem Cell Initiative, Columbia University Irving Medical Center, New York, NY 10032, USA; Initiative for Columbia Ataxia and Tremor, Columbia University Irving Medical Center, New York, NY 10032, USA; Corresponding authorSummary: Despite being among the largest neurons in the mammalian brain, Purkinje cells are difficult to visualize and trace via immunofluorescence because their dendritic arbors extend through several cerebellar layers. This protocol describes a two-antibody strategy we developed to study Purkinje cell morphology in mice. With it, one can reconstruct three-dimensional images of Purkinje cells at single-neuron resolution across multiple layers. The substantially improved image quality reveals subtle defects, enabling more meaningful morphological analysis.For complete details on the use and execution of this protocol, please refer to Gennarino et al. (2015). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722003070GeneticsMicroscopyNeuroscience |
| spellingShingle | Salvatore Botta Alexei Chemiakine Vincenzo A. Gennarino Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers STAR Protocols Genetics Microscopy Neuroscience |
| title | Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers |
| title_full | Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers |
| title_fullStr | Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers |
| title_full_unstemmed | Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers |
| title_short | Dual antibody strategy for high-resolution imaging of murine Purkinje cells and their dendrites across multiple layers |
| title_sort | dual antibody strategy for high resolution imaging of murine purkinje cells and their dendrites across multiple layers |
| topic | Genetics Microscopy Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722003070 |
| work_keys_str_mv | AT salvatorebotta dualantibodystrategyforhighresolutionimagingofmurinepurkinjecellsandtheirdendritesacrossmultiplelayers AT alexeichemiakine dualantibodystrategyforhighresolutionimagingofmurinepurkinjecellsandtheirdendritesacrossmultiplelayers AT vincenzoagennarino dualantibodystrategyforhighresolutionimagingofmurinepurkinjecellsandtheirdendritesacrossmultiplelayers |