Journal
/
Neuroscience
/
Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK
JoVE Journal
Neuroscience
This content is Free Access.
JoVE Journal Neuroscience
Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK

Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK

DOI:
10.3791/60294-v

•

11:20 min

•

December 19, 2019

•

, , , , ,
1Institute of Neurobiology,Heinrich Heine University Düsseldorf, 2Department of Neurology,Düsseldorf University Hospital

Chapters

  • 00:05Title
  • 01:25Preparation of Organotypic Brain Slice Cultures (OTCs)
  • 02:58Culturing the Slices
  • 04:18Removal of the Glial Scar
  • 05:20FRET-based ATP Imaging and Documentation of Cellular ATeam Fluorescence
  • 08:21Results: Visualization of Neurons and Astrocytes
  • 10:25Conclusion

Summary

Automatic Translation

Automatic Translation

We describe a protocol for cell-type specific expression of the genetically encoded FRET-based sensor ATeam1.03YEMK in organotypic slice cultures of the mouse forebrain. Furthermore, we show how to use this sensor for dynamic imaging of cellular ATP levels in neurons and astrocytes.

Tags

Keywords: Intracellular ATPMouse BrainFRET-based SensorATeam1.03YEMKFluorescent ImagingATP MeasurementsNeuronal ATPAstrocytic ATPEnergy MetabolismIschemic StrokeHippocampal FormationSterile ConditionsPetri DishesHanks’ Salt Solution

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/60294?article_type=v

Time-lapse Imaging of Neuroblast Migration in Acute Slices of the Adult Mouse Forebrain
check_url/60294?article_type=v

Paired Whole Cell Recordings in Organotypic Hippocampal Slices
check_url/60294?article_type=v

Regioselective Biolistic Targeting in Organotypic Brain Slices Using a Modified Gene Gun
check_url/60294?article_type=v

Imaging Serotonergic Fibers in the Mouse Spinal Cord Using the CLARITY/CUBIC Technique
check_url/60294?article_type=v

Preparation of Acute Brain Slices Using an Optimized N-Methyl-D-glucamine Protective Recovery Method
check_url/60294?article_type=v

In-depth Physiological Analysis of Defined Cell Populations in Acute Tissue Slices of the Mouse Vomeronasal Organ
check_url/60294?article_type=v

Recording Synaptic Plasticity in Acute Hippocampal Slices Maintained in a Small-volume Recycling-, Perfusion-, and Submersion-type Chamber System
check_url/60294?article_type=v

Visualization and Live Imaging of Oligodendrocyte Organelles in Organotypic Brain Slices Using Adeno-associated Virus and Confocal Microscopy
check_url/60294?article_type=v

Two-photon Imaging of Microglial Processes’ Attraction Toward ATP or Serotonin in Acute Brain Slices
check_url/60294?article_type=v

Labeling and Imaging of Amyloid Plaques in Brain Tissue Using the Natural Polyphenol Curcumin

Read Article