I'm using the doccol's 20mm silicon suture for MCAO in 20-23g of mice.
And I'm sure that I insert the suture in right place, ICA,
because the phenotype of the mouse appears right after the surgery.
However, about 2 hours after surgery, the mouse's activity is recovered.
The occlusion is not enough to study for my job.
There is anyone who can give me some tips for MCAO?
How long of an occlusion duration are you using, i.e. how long are you blocking the ICA/MCA intersection? In our lab, we regularly see full mobility within a few hours after a 30 minute occlusion. That doesn't mean the occlusion isn't working, though. There is noticeable brain damage, easily identified with TTC staining, and locomotor deficits after a 30min occlusion. However, these deficits and brain damage diminish considerably over time, and are very difficult to find after 7 days. By day 7 with TTC, you are likely to only see penumbra. Simply increasing the duration of stroke should solve your problem, assuming your experiment permits the change. However, post-op mortality rates are proportionate to the duration of stroke, so you will likely need to expand post-op care.
Also, how do you verify proper occlusion outside of the standard phenotype? Ideally, you should use a laser Doppler flowmeter to measure cerebral blood flow (CBF). Temporal CBF should be reduced by >85% to attained an adequate MCA occlusion.
First of all, I would like to thank you for your reply and I will answer your questions.
Our lab's occlusion time is 1hour and 20 minutes. I can see the locomotor deficits after the surgery.
1 day after the MCAO surgery, I confirmed with TTC staining whether the experiment was done correctly. I attached the picture of my experiment.
Our lab does not have Doppler flowmeter, so we just confirm with phenotype and TTC staining.
In summary, I'd like to expand the infarction area without adjusting the occlusion time.