This protocol has the name of monitoring in-vivo myelin chains by PET imaging with PIB labeled with carbonyl in the lysolecithin rat model of multiple sclerosis. This imaging technique has the ability of allowing in-vivo images of positions in different points. Getting point touch of the data of mining chains over time.
In this video, We show how to induce the animal model by stereotactic injection and how to acquire and analyze. The mining PIP images. Take out needed material for the lysolecithin solution preparation Take the lysolecithin solution weight in an electronic balance using a glass tube take the glass tube out from the electric balance open the cell line and take the right volume for prepare 1%lysolecithin solution.
Put the cell line in the plastic tube close the plastic tube and shake it using your fingers. Prepare for the surgery with the animal in the right position inject Put the eye cream the animal eyes are protecting against the hydration. use a 5%codoxygen solution for cleaning the incision area of the surgery inject lidocaine where the incision will be made.
Shave the hair in the heads of their skull make an incision of about two centimeters expose the scope era with udak clamps clean area with a swab and mark the bregma position the hemato syringe on the bregma injection will be made in this three marked areas through in the right coordinate. Take the hemato needle through the drute area Go to the ventrical coordinate for start the injection, Finishing the surgery, Suture the incision. Use an appropriate place for the tracer injection.
The tracer must be injected intravenously, and it can be done either, In the penile vein or in the tail vein. Image acquisition of baseline time point is performed before stereotactic surgery. And the other two time points one week and four weeks after the stereotactic surgery.
Open the PET scanner software and chose the field of view to be scanned during image acquisition. Position the animal in the bed. Be sure that the animal is straight.
Take the eye cream to protect the animal eyes. Cover the animal bed. Push the animal bed inside the pet scanner and start acquisition.
Check the animal griefing until the end of the PET scan. When one day image acquisition is finished. Take animal out of the bed.
Open remote software and choose Fuse It'model select image bio at and open with operations. Use a cylinder orientation and load the image. Go to matching and open the PET image.
Adjust color scale to make the brain visible. Press crop'to crop the image and adjust the window size to cover the whole brain. It's parameters are on the bottom right side of the screen press blue shift'Be sure that the animal is positioned right in the software.
Select the correct MRI template. if you need it, you can adjust the coverage restriction between PET and MRI template. Press device'button.
Select the right brain template. Then you can look at different brain regions available and you can select or unselect all of them. Go to outline guides, and they will be all selected.
In the top button, you can select any parameter you want to appear in this statistic data. You can touch the statistic button and then you can choose between KilobeckerCSS or SUV. Copy to clipboard and close.
Your data can be pasted in an Excel sheet. For quantifying the and delusion Go to the new board, select these few and apply. You can create a new void in the contralateral side.
And then go to the statistic button S2B4. It is possible to see the lesion one week after injection and the lesions covered four weeks after injection. Significant difference are seen between cell lining one week and four.
Following this protocol, you'll get the three steps of taking the region of interest, being the injection site and that the principle area to be analyzed. As a large student of protocol, thus taking lesion presents the mining chains over time, where the mining nation is represented by decreased trace uptake and reimagination trace uptake. It's important to emphasize that the laser listing injection must be very slow, avoiding tissue damaging and causing only mining chains which will be detect by peep PIP imaging.
