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A perfusion protocol for lizards, including a method for brain removal.

Hoops D - MethodsX (2015)

Bottom Line: The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state.Increasingly, other vertebrate groups are also being used in neuroscience.Though geared towards standard brain perfusion, this protocol is easily modified for the perfusion of other tissues and for various specialized histological techniques. •The two aortas of the lizard heart, emerging from a single ventricle, mean that care must be taken to place the perfusion needle in the correct aorta, unlike in mammals.•Only the head and neck perfuse - the visceral organs will not decolour, and the body may not twitch.•I also include a method for removing a lizard brain, which differs from mammals due to the incomplete and thicker skull of the lizard.

View Article: PubMed Central - PubMed

Affiliation: Evolution, Ecology & Genetics, Research School of Biology, The Australian National University, 116 Daley Road, Acton, ACT 2601, Australia.

ABSTRACT
The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. Perfusion achieves optimal fixation by pumping fixative directly through an animal's circulatory system. Standard perfusion techniques were developed primarily for application in mammals, which are traditional neuroscience research models. Increasingly, other vertebrate groups are also being used in neuroscience. Following mammalian perfusion protocols for non-mammalian vertebrates often results in failed perfusions. Here, I present a modified perfusion protocol suitable for lizards. Though geared towards standard brain perfusion, this protocol is easily modified for the perfusion of other tissues and for various specialized histological techniques. •The two aortas of the lizard heart, emerging from a single ventricle, mean that care must be taken to place the perfusion needle in the correct aorta, unlike in mammals.•Only the head and neck perfuse - the visceral organs will not decolour, and the body may not twitch.•I also include a method for removing a lizard brain, which differs from mammals due to the incomplete and thicker skull of the lizard.

No MeSH data available.


Related in: MedlinePlus

Lateral, dorsal, and caudal views of a standardized lizard skull which highlight the bones that must be removed to extract the brain. Prior to the removal of the skull, the head should be pinned through the external nares and upper temporal fenestras (step 5.1). First, the postorbital bone is cut, represented by the red lines (step 5.2). Second, the spine, shown in purple, is removed (step 5.4). Third, the posterior skull, in green, is removed (step 5.6). Fourth, the dorsal skull, in orange, is removed (steps 5.7–5.8). Finally, the ear bones, in yellow, are removed (step 5.11).
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fig0020: Lateral, dorsal, and caudal views of a standardized lizard skull which highlight the bones that must be removed to extract the brain. Prior to the removal of the skull, the head should be pinned through the external nares and upper temporal fenestras (step 5.1). First, the postorbital bone is cut, represented by the red lines (step 5.2). Second, the spine, shown in purple, is removed (step 5.4). Third, the posterior skull, in green, is removed (step 5.6). Fourth, the dorsal skull, in orange, is removed (steps 5.7–5.8). Finally, the ear bones, in yellow, are removed (step 5.11).

Mentions: 4.15 Pin the head to the dissection pad (Fig. 3A). Two pins are inserted through the external nares and two more pins through the upper temporal fenestras (Fig. 4).


A perfusion protocol for lizards, including a method for brain removal.

Hoops D - MethodsX (2015)

Lateral, dorsal, and caudal views of a standardized lizard skull which highlight the bones that must be removed to extract the brain. Prior to the removal of the skull, the head should be pinned through the external nares and upper temporal fenestras (step 5.1). First, the postorbital bone is cut, represented by the red lines (step 5.2). Second, the spine, shown in purple, is removed (step 5.4). Third, the posterior skull, in green, is removed (step 5.6). Fourth, the dorsal skull, in orange, is removed (steps 5.7–5.8). Finally, the ear bones, in yellow, are removed (step 5.11).
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4487709&req=5

fig0020: Lateral, dorsal, and caudal views of a standardized lizard skull which highlight the bones that must be removed to extract the brain. Prior to the removal of the skull, the head should be pinned through the external nares and upper temporal fenestras (step 5.1). First, the postorbital bone is cut, represented by the red lines (step 5.2). Second, the spine, shown in purple, is removed (step 5.4). Third, the posterior skull, in green, is removed (step 5.6). Fourth, the dorsal skull, in orange, is removed (steps 5.7–5.8). Finally, the ear bones, in yellow, are removed (step 5.11).
Mentions: 4.15 Pin the head to the dissection pad (Fig. 3A). Two pins are inserted through the external nares and two more pins through the upper temporal fenestras (Fig. 4).

Bottom Line: The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state.Increasingly, other vertebrate groups are also being used in neuroscience.Though geared towards standard brain perfusion, this protocol is easily modified for the perfusion of other tissues and for various specialized histological techniques. •The two aortas of the lizard heart, emerging from a single ventricle, mean that care must be taken to place the perfusion needle in the correct aorta, unlike in mammals.•Only the head and neck perfuse - the visceral organs will not decolour, and the body may not twitch.•I also include a method for removing a lizard brain, which differs from mammals due to the incomplete and thicker skull of the lizard.

View Article: PubMed Central - PubMed

Affiliation: Evolution, Ecology & Genetics, Research School of Biology, The Australian National University, 116 Daley Road, Acton, ACT 2601, Australia.

ABSTRACT
The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. Perfusion achieves optimal fixation by pumping fixative directly through an animal's circulatory system. Standard perfusion techniques were developed primarily for application in mammals, which are traditional neuroscience research models. Increasingly, other vertebrate groups are also being used in neuroscience. Following mammalian perfusion protocols for non-mammalian vertebrates often results in failed perfusions. Here, I present a modified perfusion protocol suitable for lizards. Though geared towards standard brain perfusion, this protocol is easily modified for the perfusion of other tissues and for various specialized histological techniques. •The two aortas of the lizard heart, emerging from a single ventricle, mean that care must be taken to place the perfusion needle in the correct aorta, unlike in mammals.•Only the head and neck perfuse - the visceral organs will not decolour, and the body may not twitch.•I also include a method for removing a lizard brain, which differs from mammals due to the incomplete and thicker skull of the lizard.

No MeSH data available.


Related in: MedlinePlus