Transcardial perfusion fixation of the CNS by means of a compressed-air-driven device

Jonkers, B.W.; Sterk, J.; Wouterlood, Floris G.

doi:10.1016/0165-0270(84)90013-x

Cited by 34 publications

(15 citation statements)

References 7 publications

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“…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…”

mentioning

confidence: 99%

Whole Animal Perfusion Fixation for Rodents

Gage

Kipke

Shain

2012

JoVE

419

315

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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. Video LinkThe video component of this article can be found at https://www.jove.com/video/3564/ Protocol

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mentioning

confidence: 99%

Whole Animal Perfusion Fixation for Rodents

Gage

Kipke

Shain

2012

JoVE

419

315

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“…The techniques described may be used with little or no adaptation in landrace pigs and other large research animals such as dogs and sheep 14 . Aseptic technique must be employed to avoid infections, when performing the described procedures in survival studies.…”

Section: Discussionmentioning

confidence: 99%

Basic Surgical Techniques in the Göttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

Ettrup

Glud

Orłowski

et al. 2011

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The emergence of the Göttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects the close resemblance of these animals to human anatomy and physiology [1][2][3][4][5][6] .The size of the Göttingen minipig permits the use of surgical equipment and advanced imaging modalities similar to those used in humans [6][7][8] . The aim of this instructional video is to increase the awareness on the value of minipigs in biomedical research, by demonstrating how to perform tracheal intubation, transurethral bladder catheterization, femoral artery and vein catheterization, as well as transcardial perfusion.Endotracheal Intubation should be performed whenever a minipig undergoes general anesthesia, because it maintains a patent airway, permits assisted ventilation and protects the airways from aspirates. Transurethral bladder catheterization can provide useful information about about hydration state as well as renal and cardiovascular function during long surgical procedures. Furthermore, urinary catheterization can prevent contamination of delicate medico-technical equipment and painful bladder extension which may harm the animal and unnecessarily influence the experiment due to increased vagal tone and altered physiological parameters. Arterial and venous catheterization is useful for obtaining repeated blood samples and monitoring various physiological parameters. Catheterization of femoral vessels is preferable to catheterization of the neck vessels for ease of access, when performing experiments involving frame-based stereotaxic neurosurgery and brain imaging. When performing vessel catheterization in survival studies, strict aseptic technique must be employed to avoid infections 6 . Transcardial perfusion is the most effective fixation method, and yields preeminent results when preparing minipig organs for histology and histochemistry 2,9

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“…The animals were overdosed with ketamine (50 mg/kg)/ xylazine (0.024 mg/kg)/acepromazine (0.005 mg/kg) cocktail and transcardially perfused with phosphate buffer saline (PBS) followed by 4% PFA as previously described [192]. The spinal cord including the injury epicenter and lumbar enlargement was dissected out and post-fixed in 4% PFA for one hour and transferred to 30% sucrose for cryoprotection for at least 4 days.…”

Section: Sacrifice and Tissue Processingmentioning

confidence: 99%

“…Animals were overdosed with a ketamine (50 mg/kg)/ xylazine (0.024 mg/kg)/ acepromazine (0.005 mg/kg) cocktail and transcardially perfused with phosphate buffer, followed by 4% paraformaldehyde (PFA) [192]. The spinal cord and hindlimb muscles (Biceps Femoris, Gastrocnemius and Tibialis Anterior) were dissected out, post-fixed in 4% PFA overnight and cryprotected with 30% sucrose.…”

Section: Histological Proceduresmentioning

confidence: 99%

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Stretching adversely modulates locomotor capacity following spinal cord injury via activation of nociceptive afferents.

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Transcardial perfusion fixation of the CNS by means of a compressed-air-driven device

Cited by 34 publications

References 7 publications

Whole Animal Perfusion Fixation for Rodents

Whole Animal Perfusion Fixation for Rodents

Basic Surgical Techniques in the Göttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

Stretching adversely modulates locomotor capacity following spinal cord injury via activation of nociceptive afferents.

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Transcardial perfusion fixation of the CNS by means of a compressed-air-driven device

Cited by 34 publications

References 7 publications

Whole Animal Perfusion Fixation for Rodents

Whole Animal Perfusion Fixation for Rodents

Basic Surgical Techniques in the G&ouml;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

Stretching adversely modulates locomotor capacity following spinal cord injury via activation of nociceptive afferents.

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Basic Surgical Techniques in the Göttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion