Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 65, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/3564
Keywords
Neuroscience; Issue 65; Physiology; Biomedical Engineering; Paraformaldehyde; fixation; physiological pressures; vascular system; brain; preservation
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Funding
- Center for Neural Communication Technology (CNCT)
- National Institute of Biomedical Imaging and Bioengineering (NIBIB) [P41 EB002030]
- National Institutes of Health (NIH)
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The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small pieces of tissue, larger specimens like the intact brain pose a problem for immersion fixation because the fixative does not reach all regions of the tissue at the same rate (5,7). Often, changes in response to hypoxia begin before the tissue can be preserved (12). The advantage of directly perfusing fixative through the circulatory system is that the chemical can quickly reach every corner of the organism using the natural vascular network. In order to utilize the circulatory system most effectively, care must be taken to match physiological pressures (3). It is important to note that physiological pressures are dependent on the species used. Techniques for perfusion fixation vary depending on the tissue to be fixed and how the tissue will be processed following fixation. In this video, we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain for immunohistochemistry. The main advantage of this technique (vs. gravity-fed systems) is that the circulatory system is utilized most effectively.
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