Monitoring neuronal survival via longitudinal fluorescence microscopy

Kaitlin Weskamp, Nathaniel Safren, Roberto Miguez, Sami Barmada*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

Standard cytotoxicity assays, which require the collection of lysates or fixed cells at multiple time points, have limited sensitivity and capacity to assess factors that influence neuronal fate. These assays require the observation of separate populations of cells at discrete time points. As a result, individual cells cannot be followed prospectively over time, severely limiting the ability to discriminate whether subcellular events, such as puncta formation or protein mislocalization, are pathogenic drivers of disease, homeostatic responses, or merely coincidental phenomena. Single-cell longitudinal microscopy overcomes these limitations, allowing the researcher to determine differences in survival between populations and draw causal relationships with enhanced sensitivity. This video guide will outline a representative workflow for experiments measuring single-cell survival of rat primary cortical neurons expressing a fluorescent protein marker. The viewer will learn how to achieve high-efficiency transfections, collect and process images enabling the prospective tracking of individual cells, and compare the relative survival of neuronal populations using Cox proportional hazards analysis.

Original languageEnglish (US)
Article numbere59036
JournalJournal of Visualized Experiments
Volume2019
Issue number143
DOIs
StatePublished - Jan 2019
Externally publishedYes

Keywords

  • Automation
  • Cell death
  • Fluorescence microscopy
  • Issue 143
  • Neurodegeneration
  • Neuroscience
  • Survival analysis
  • Transfection

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Fingerprint

Dive into the research topics of 'Monitoring neuronal survival via longitudinal fluorescence microscopy'. Together they form a unique fingerprint.

Cite this