|
Title
|
|
|
|
Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation
| |
|
Author
|
|
|
|
| |
|
Abstract
|
|
|
|
| Neurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmentalize single synapses to fine-tune synaptic strength. However, accurate quantification of spine density and morphology in mature neuronal networks is hampered by the lack of targeted labeling strategies. To resolve this, we have optimized a method to deliver cell-impermeable compounds into selected cells based on Spatially resolved NAnoparticle-enhanced Photoporation (SNAP). We show that SNAP enables efficient labeling of selected individual neurons and their spines in dense cultured networks without affecting short-term viability. We compare SNAP with widely used spine labeling techniques such as the application of lipophilic dyes and genetically encoded fluorescent markers. Using SNAP, we demonstrate a time-dependent increase in spine density in healthy cultures as well as a reduction in spine density after chemical mimicry of hypoxia. Since the sparse labeling procedure can be automated using an intelligent acquisition scheme, SNAP holds promise for high-content screening campaigns of neuronal connectivity in the context of neurodevelopmental and neurodegenerative disorders. |
| |
|
Language
|
|
|
|
English
| |
|
Source (journal)
|
|
|
|
Frontiers in cellular neuroscience
| |
|
Publication
|
|
|
|
2018
| |
|
ISSN
|
|
|
|
1662-5102
| |
|
DOI
|
|
|
|
10.3389/FNCEL.2018.00080
| |
|
Volume/pages
|
|
|
|
12
(2018)
, 11 p.
| |
|
Article Reference
|
|
|
|
80
| |
|
ISI
|
|
|
|
000428644200001
| |
|
Pubmed ID
|
|
|
|
29651235
| |
|
Medium
|
|
|
|
E-only publicatie
| |
|
Full text (Publisher's DOI)
|
|
|
|
| |
|
Full text (open access)
|
|
|
|
| |
|