Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays
Brain development is highly complex and dynamic. During this process, the different brain structures acquire new components, such as the cerebral cortex, which builds up different germinal and cortical layers during its development. The genetic study of this complex structure has been commonly appro...
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Bio-protocol LLC
2024-12-01
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| Series: | Bio-Protocol |
| Online Access: | https://bio-protocol.org/en/bpdetail?id=5133&type=0 |
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| author | Lucia Del-Valle-Anton Salma Amin Víctor Borrell |
| author_facet | Lucia Del-Valle-Anton Salma Amin Víctor Borrell |
| author_sort | Lucia Del-Valle-Anton |
| collection | DOAJ |
| description | Brain development is highly complex and dynamic. During this process, the different brain structures acquire new components, such as the cerebral cortex, which builds up different germinal and cortical layers during its development. The genetic study of this complex structure has been commonly approached by bulk-sequencing of the entire cortex as a whole. Here, we describe the methodology to study this layered tissue in all its complexity by microdissecting two germinal layers at two developmental time points. This protocol is combined with a step-by-step explanation of tissue dissociation that provides high-quality cells ready to be analyzed by the newly developed single-cell assays, such as scRNA-seq, scATAC-seq, and TrackerSeq. Altogether, this approach increases the resolution of the genetic analyses from the cerebral cortex compared to bulk studies. It also facilitates the study of laboratory animal models that recapitulate human cortical development better than mice, like ferrets. |
| format | Article |
| id | doaj-art-04a019f6d8c74f1eb04ac1c266a8f51b |
| institution | Kabale University |
| issn | 2331-8325 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Bio-protocol LLC |
| record_format | Article |
| series | Bio-Protocol |
| spelling | doaj-art-04a019f6d8c74f1eb04ac1c266a8f51b2025-02-07T08:16:22ZengBio-protocol LLCBio-Protocol2331-83252024-12-01142410.21769/BioProtoc.5133Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell AssaysLucia Del-Valle-Anton0Salma Amin1Víctor Borrell2Instituto de Neurociencias, Consejo Superior de Investigaciones Científicas & Universidad Miguel Hernández, Sant Joan d’Alacant, SpainInstituto de Neurociencias, Consejo Superior de Investigaciones Científicas & Universidad Miguel Hernández, Sant Joan d’Alacant, SpainInstituto de Neurociencias, Consejo Superior de Investigaciones Científicas & Universidad Miguel Hernández, Sant Joan d’Alacant, SpainBrain development is highly complex and dynamic. During this process, the different brain structures acquire new components, such as the cerebral cortex, which builds up different germinal and cortical layers during its development. The genetic study of this complex structure has been commonly approached by bulk-sequencing of the entire cortex as a whole. Here, we describe the methodology to study this layered tissue in all its complexity by microdissecting two germinal layers at two developmental time points. This protocol is combined with a step-by-step explanation of tissue dissociation that provides high-quality cells ready to be analyzed by the newly developed single-cell assays, such as scRNA-seq, scATAC-seq, and TrackerSeq. Altogether, this approach increases the resolution of the genetic analyses from the cerebral cortex compared to bulk studies. It also facilitates the study of laboratory animal models that recapitulate human cortical development better than mice, like ferrets.https://bio-protocol.org/en/bpdetail?id=5133&type=0 |
| spellingShingle | Lucia Del-Valle-Anton Salma Amin Víctor Borrell Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays Bio-Protocol |
| title | Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays |
| title_full | Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays |
| title_fullStr | Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays |
| title_full_unstemmed | Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays |
| title_short | Microdissection and Single-Cell Suspension of Neocortical Layers From Ferret Brain for Single-Cell Assays |
| title_sort | microdissection and single cell suspension of neocortical layers from ferret brain for single cell assays |
| url | https://bio-protocol.org/en/bpdetail?id=5133&type=0 |
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