Okay.Hello, my name is Alan Meyer and I'm a research assistant in the faf Laboratory at the Salk Institute. Today I'm going to show you how to perform a ventral laminectomy to isolate the spinal cord of a mouse as well, I will show you how to attach the spinal cord to an electrode. To record its neural activity, you'll need to prepare two solutions for the spinal cord, one for the dissection, and the other for the recording bath.
The only difference in the two solutions will be the concentration of calcium chloride. You will first need to prepare a solution of 10 X-A-C-S-F. This will require the reagents listed below.
Begin by adding 100 milliliters of the 10 X-A-C-S-F to a one liter volumetric flask. Next, fill the flask with water, but leave room so as not to overflow. When gassed gas, the solutions with a gas concentration of 5%CO2, and 95%O2.
To lower the pH and oxygenate solution, add one milliliter of one molar magnesium sulfate. Next, add one milliliter of a one molar calcium chloride solution to the dissection solution, and add two milliliters of one molar calcium chloride to the recording solution. Mice from ages E 16.5 to P four can be used in these experiments.
Begin dissecting the mouse by first swiftly, decapitating it with sharp scissors. Next, cut along the ventral midline until you reach the tail. Then using forceps, eviscerate the mouse, and rinse with the A CSF solution.
Use the one millimolar calcium chloride A CSF to perform the dissection. Place the mouse in the dissecting dish and pin the mouse out by placing a pin at its four paws and a pin at the base of the tail. Our goal for this portion Will be to isolate the spinal cord by cutting along the red dotted line, separating the spinal column from the lamina that encase the spinal cord.
This will allow you to expose the spinal cord. Next, follow along the line to cut out the meninges and spinal roots, freeing the spinal cord. Locate the spinal column and use one hand with tweezers to firmly grip the spinal column and the other with small scissors.
To remove the spinal cord, keep the scissors at a low angle. To avoid puncturing the cord with the tips of the scissors, we will now dissect the spinal cord. Following along the red line, cut the meninges and spinal roots.
To remove the spinal cord, you can cut along either side of the cord and then cut along the ventral midline. It is important to keep the scissors at a low angle so as not to puncture the spinal cord. It is important to avoid cutting the roots too close on the spinal cord.
Next, transfer the spinal cord to the recording bath and connect the electrodes to the spinal roots. Position the electrode near the root. Then using suction gently draw the nerve into the electrode.
Thank you for watching and I hope this video will be helpful in your future experiments.
