These videos demonstrate the OPTIMAL conditions and employ all WELFARE guidelines (which conveniently exclude 90% of all lab victims) that regulate animal torture in labs. Note the utter disregard for life, and the upbeat music that accompanies the mutilation. There is no shred of decency, no respect, no empathy… living beings are simply objects to these people. When the cameras stop rolling and no one is watching, innocent animals are helpless in the bloody hands of true sociopaths.
Watch Video: http://www.jove.com/index/Details.stp?ID=880
Two-month old average age C57BL/6J mice or equivalent or three-month old average age Sprague Dawley rats or equivalent are anesthetized with ketamine (60 mg/kg i.p. for rats; 100 mg/kg i.p for mice) and xylazine (10 mg/kg, i.p. for either species). Sedation is monitored using a gentle toe pinch withdraw reflex demonstrated in Walantus et al.(JoVE, 6, 2007) and Szot et al.(JoVE, 9, 2007). Thermoregulation can be provided through a thermostatregulated heating pad (ALA Instruments Inc.) and monitored through a rectal thermometer. Head is shaved of fur and cleaned with iodine before incision. After skin incision (2 cm long for rats, 1 cm long for mice) and removal of all soft tissue from the surface of the skull, placement of the guide cannula is determined in relation to bregma. A 6 mm hole is drilled through the skull with a battery-operated driller designed for rodent surgery (Fine Science Tools, Inc.). Care is taken so that the drill bit does not penetrate through meningeal membranes or blood vessels. Skull is implanted with bilateral 5 mm 21 gauge stainless steel guide shafts leading to the posterior nucleus accumbens, dorsal striatum or prefrontal cortex. The stereotaxic coordinates are established as per Franklin and Paxinos, 1997 (The Mouse Brain in Stereotaxic Coordinates, Academic Press) or Paxinos and Watson, 2006 (The Rat Brain in Stereotaxic Coordinates, Academic Press). Implants are secured by dental cement. A bolus of Lactated Ringers of the 0.9% saline is given at the end of surgery (5mls SC in rats and 1 ml SC in mice after fluids are warmed to normal body temperature) to prevent dehydration. Buprenorphine (0.1-0.5mg/kg SC) is administered twice daily and, then, on an as-needed basis, if animal appears to be in pain. Local antibiotic treatment (bacitracin ointment) and systemic antibiotic treatment (penicillin 100,000 IU/kg IM every 12 hours for the first 48 hours post-op) are administered if post-operative infections occur.
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1Department of Physiology, University of Montreal, 2Ecole d’optometrie, University of Montreal, 3Stereology Resource Center
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Department of Neurology, University of California, Los Angeles
- Anesthetize mice with isoflurane (4% for induction, 1.5-2% for surgery) using IACUC approved procedures. It is important that tail and/or toe pinches are used in order to ensure the animal is fully sedated.
- Using a rodent trimmer, shave the hair from the back of the neck up to the eyes.
- Place the mouse in a stereotaxic frame, over a surgery water re-circulating blanket. Firmly secure the head with ear bars.
- Apply eye ointment, in order to prevent the animal’s eye from drying out.
- Administer, subcutaneously, Dexamethasone (0.2 mg/Kg) and Carprofen (5 mg/Kg) to prevent swelling of the brain and/or an inflammatory response, respectively.
- Before beginning the surgery, sterilize the operating area by wiping skin with three alternating swipes of 70% alcohol and betadine.
- All surgical instruments have been pre-sterilized using a glass bead sterilizer. Using scissors that have been sterilized with ethanol, remove the skin over the top of the skull, starting with a horizontal cut all along the base of the head, followed by two cuts in the rostral direction, almost reaching the eyelids, then two oblique cuts that converge at the midline.
- A drop of lidocaine + epinephrine solution is applied at this point onto the periosteum to avoid excessive bleeding or pain. With a scalpel, retract the periosteum to the edges of the skull. Also, lightly retract the musculature of the back of the neck.
- Gently scrape the entire exposed area of the skull with the scalpel to create a dry surface. This is very important, as it will allow the glue to adhere better when applied later.
- Once an imaging site has been chosen, one is ready to create the cranial window. First, gently “draw” a circle of about 4 mm in diameter with the pneumatic dental drill.
- After a slight drilling, apply lidocaine + epinephrine solution again onto the skull surface. Stop the drilling when a very thin layer of bone is left. By pushing gently on the center of the craniotomy to feel how it gives way, one can usually know that this stage is reached.
- Under a drop of saline and taking advantage of the bone trabeculae – the spongy structure of the bone – lift away the craniotomy from the skull with very thin tip forceps. The saline is important, as it will help lift up the skull and prevent bleeding of the dura.
- Apply Gelfoam that has been previously soaked in saline to the dura mater to stop any small bleeding that occassionally occurs when the skull is removed.
- After drying the dura mater surface and ensuring that there is no bleeding, gently lay a sterile 5 mm glass cover slip on top of the dura mater. (Note: other groups also place a drop of low melting point agarose (1.2%) over the dura and put the coverslip on top of the agar).
- Apply a drop of cyanocrylate-based glue to the opposite hemisphere on the skull. With the help of a needle, gently apply the glue all around the window while being careful not to put it under the glass. Glue can now be applied in a thin layer over the entire surface of the skull.
- Once the glue has dried, mix dental acrylic and apply it throughout the skull surface, covering also a small rim of the cover slip, to secure it.
- After securing the cover slip, make a small well around the window with dental acrylic. Also, embed a titanium bar in the dental acrylic. This bar will later be used to attach the mouse securely on to the stage of the microscope for imaging. It is important to ensure that the bar is level, so that it is parallel with the cranial window. Placing a piece of paper under the bar can allow the bar to remain level while the acrylic hardens.
- The dental acrylic is allowed to cure (harden) for 10 minutes, by which time the titanium bar is fixed in place. Place the animal in a warm cage until it recovers.
- After recovery from anesthesia, the animal can be imaged on the same day.
1Department of Physiology, University of Montreal, 2School of Optometry, University of Montreal, 3Departement de chimie-biologie, Universite du Quebec a Trois-Rivieres
Part 1: Pre-processing of tissue
- Tissue should be well perfused with paraformaldehyde, glutaraldehyde, or formalin. This can be achieved through standard transcardial perfusion typically used to harvest other organs. It is recommended that shortly after sacrifice the eyes be injected with fixative just under the lens and stored in fixative.
- In the present study the subject was deeply sedated with ketamine hydrochloride (10 mg/kg, i.m.), euthanized with an overdose of sodium pentobarbital (25 mg/kg, i.v.) and perfused transcardially with 0.1 M PBS until completely exsanguinated. This is followed by a 4% paraformaldehyde solution in PBS for 5 min (~1 liter).
Part 2: Removal of the eyeball from the orbital cavity
- For easier access to the eyeball it is recommended to first remove the brain. Once the brain has been removed the thin-walled orbit bone is readily apparent. Use the bone rongeurs to slowly chip away the wall of the orbit. Cut away the ocular muscles with a scalpel and remove the connective tissue from the eyeball. Carefully cut the optic nerve, this can be used in electronic microscopy studies. The eyeball should now be released from the orbital cavity.
Part 3: Dissect the retina from the eyecup
- Place the eye into a Petri dish with PBS to keep the retina from drying. A dissecting microscope or table mounted magnifying light stand is useful in the dissection, but not a necessity. Remove the cornea by cutting the sclera closely to the perimeter of the cornea at the level of the ora serrata with a pair of spring scissors and remove the lens with the forceps.
- Use a paintbrush, forceps, and spring scissors to remove the retina from the sclera. This is done by separating the retina from the sclera and then cutting the sclera away with the scissors. One must carry out this process in small increments so as not to damage or tear the retinal tissue. The sclera is not readily separated from the remnant optic nerve so carefully cut the sclera around the optic nerve.
- At this point the retina has retained its curved shape and needs to be flattened for sampling. Before flattening the retina, the vitreous humour, which has the consistency of jelly, can be removed in a lump. To flatten the retina onto a slide, make several radial cuts with a scalpel blade. The residual vitreous humour can now be removed with ordinary filter paper and a paintbrush. The retinal ganglion cell layer is exposed at this point so it is imperative to be gentle when removing the vitreous humour. If the optic nerve is still attached at the optic disc, remove the optic nerve without ripping the retina using a scalpel blade and a pair of spring scissors. This is now a flat mount retina (Figure 1) and the fovea should be apparent as a dark patch in the temporal/ventral direction from the optic disc.
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What do you expect? For them to be crying while talking or maybe hold a funeral? You call it ‘mutilation’. I call it surgery. Welfare guidelines look like they’re being met, you do know the difference between welfare and rights, yeah?
“Surgery” is a gross euphemism that implies necessity.
Vivisection is mutilation with no consideration given to the victim that is being tortured.
Welfare is an abomination that simply regulates the atrocities.
You are an ignorant speciesist.
The problem is that animal welfare is a construct we have created to tell ourselves that our use of animals is ethically justifiable. This is akin to the slaveholder claiming that there’s no fundamental problem with slavery, so long as he’s relatively kind (by his own standards) to his slaves.
The actual interests of the slaves are never really considered, at all.
We decide what animal welfare is, who gets it, what it entails, when to apply it, when *not* to apply it, and so on.
The whole construct is just humans telling themselves that animals are an exploitable resource – but it’s okay, because we do it “nicely” (with standards of “nice treatment” as defined by us).