Home › Microscopy & Imaging › Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
Microscopy & Imaging JoVE (Open Access) Citable · DOI

Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition

DOI: 10.3791/56282-v
What you'll learn
  • ✓ Assemble and operate a perfusion system for maintaining acute brain slice viability
  • ✓ Isolate and position hippocampal and cortical slices for MR microscopy imaging
  • ✓ Acquire high-quality magnetic resonance microscopy data from living tissue preparations
  • ✓ Evaluate tissue preservation and image quality during prolonged MR data collection
Protocol

The current protocol describes a method by which users can maintain viability of acute hippocampal and cortical slice preparations during the collection of magnetic resonance microscopy data.

Difficulty
advanced
Total time
~3–4 hours per animal (brain extraction, slice isolation, perfusion setup, and MR acquisition)
Model organism
Mouse (unspecified strain, likely C57BL/6 or similar laboratory strain)
Biosafety
BSL-1

Steps

1
Assemble perfusion system hardware and tubing

Set up the perfusion apparatus including pump, reservoir, heating module, and connected tubing. Prepare artificial cerebrospinal fluid (aCSF) and prime the system to ensure continuous, temperature-controlled delivery.

▶ 00:53
2
Extract and prepare acute brain tissue slices

Perform rapid brain extraction and prepare acute hippocampal and cortical slices using standard slicing techniques. Maintain tissue viability through cold, oxygenated aCSF during isolation.

▶ 03:31
3
Position slice in perfusion chamber for imaging

Mount the isolated brain slice in the perfusion chamber, align it within the MR microscopy coil, and connect the perfusion system to initiate continuous aCSF delivery during acquisition.

▶ 04:55
4
Collect magnetic resonance microscopy images

Execute MR microscopy pulse sequences to acquire high-resolution structural images of the perfused brain slice while maintaining metabolic support throughout the scan.

▶ 06:39
5
Evaluate tissue viability and image quality

Assess the quality of acquired MR images and evaluate tissue preservation and morphology to confirm successful maintenance of slice viability during the imaging protocol.

▶ 07:48
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