Recent Posts

31

Surgery and Baseplating / Re: nothing but gray

« Last post by hsiao on August 16, 2018, 05:24:32 AM »

But rat surgery is another story.
I feel like the 1st layer of CC of rats are tough and thick. (plz see the Rat dCA1_1.JPG and Rat dCA1_2.JPG)
After removing them, I saw some thin and transparent layers (plz see the Rat dCA1_2.JPG and Rat dCA1_3.JPG) so I think they are vertical CC and the alveus is below them.
Maybe this is a good place to implant lens?

32

Surgery and Baseplating / Re: nothing but gray

« Last post by hsiao on August 15, 2018, 08:32:35 AM »

I did a practice surgery on mouse and rat targeting dCA1 and noticed some difference.

Thank you, Daniel, for reminding me of the tutorial videos.

For mice
Yes, everything just like the videos regarding the aspiration.
I saw two layers of the corpus callosum and I saw the fiber direction and blood vessels clearly under the stereoscope.
Using aspiration tools can easily remove the 1st layer of CC.
I am very excited about seeing this. I also took a picture of it (plz see the attached picture).

33

Surgery and Baseplating / Dental Cement for Two Lens System

« Last post by Mark Benz on August 14, 2018, 05:26:34 PM »

Hello,
 
My lab will be using a two lens system to image the mPFC. We are currently in the initial stages of the first surgery (implanting the relay lens in the brain). This requires the we cement the relay lens to the skull. However, we are having trouble finding a fast curing, hard dental cement. We purchased the products recommended by the following protocol:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5239057/

Though we are finding that the Lang Dental products are quite malleable after curing and also set quite slowly. Has anyone had success with Lang Dental's Jet Denture Repair Powder and Ortho-Jet Crystal Liquid? If so, is there anything that we might be doing wrong? Currently, we just mix the liquid and powder and then let dry.

Alternately, are there other products that you've found work well for these types of surgeries?

Any help is great appreciated!

best wishes,
Mark Benz (Jackman Lab, OHSU)

34

Surgery and Baseplating / Re: nothing but gray

« Last post by Daniel Aharoni on August 10, 2018, 02:40:43 AM »

Hi hsiao,
You need to always use the objective GRIN lens in conjunction with the relay lens. If you try to image with just the relay lens you will never see anything in focus.

1. You remove the corpus callosum with aspiration, never forceps. I think we should have videos in the surgery power point presentation on miniscope.org under Workshop Resources.
2. You should be doing all your aspiration under a steroscope. You will visually see the fibers turn direction as you aspirate deeper.
3. We generally do not see much during implantation. It takes a could weeks for the imaging field to clear up to see a decent image.

35

Surgery and Baseplating / Re: nothing but gray

« Last post by hsiao on August 09, 2018, 03:22:13 PM »

Still have no luck on my second rat ,

3 weeks after virus injection, I check the image today.

This time, I did not use the objective lens.
The relay lens just sticks out the skull about 1mm.

I think I spotted a thin blood vessel (not sure since it is not clear either) but could not find any white neuron.
I cemented the baseplate anyway because I think maybe the cells will appear next week.


Maybe it is something wrong during the surgery...

I read the instruction again and found one step that I didn't do: Very slowly and carefully remove the horizontal white striations of the corpus callosum until you reach the striations that go vertical. 
Several questions I want to ask about it.

1. How do you remove the corpus callosum? by tweezer, needle or aspiration?
2. How to tell the horizontal and vertical corpus callosum?
3.
During the implantation surgery, will you see blood vessels through the relay lens?
I didn't see any blood vessel on the corpus callosum during the surgery. Is that normal?
Maybe I can't see the blood vessels during implantation and baseplating it is due to lack of removing corpus callosum?
I think it may be good to have a same blood vessel landmark during surgery and baseplating.

Thank you

36

Miniscope.org Suggestions / Re: Member pages broken

« Last post by Daniel Aharoni on August 09, 2018, 02:00:58 AM »

Thanks. I'll try and look into this soon.

37

Surgery and Baseplating / Re: nothing but gray

« Last post by TBNentwig on August 04, 2018, 06:04:49 AM »

Kristen,

What brain region are you targeting?

From what I have heard from people trying to image deep brain regions in rats (here at MUSC) using the Inscopix system is that it is very difficult and they have been unsuccessful. They have got good cells in mPFC but it seems like deep targets like NAc and VP don't yield good cells, not sure of the theory behind it, perhaps due to more inflammation in rats.

38

Miniscope.org Suggestions / Member pages broken

« Last post by Georgetown34 on August 03, 2018, 08:25:05 PM »

Hi,

My lab was looking to set up our member page and put up info on how we've built our scopes. However, I can't access the member pages or even my own. When I click on them it gives me a 403 Forbidden error.

Just a heads up so that can be fixed.

Thanks.

39

Surgery and Baseplating / Re: nothing but gray

« Last post by hsiao on August 01, 2018, 03:30:14 PM »

Hi Kristen,

Do you mind to tell me the information from Tad Blair's lab?

Thanks!

Hsiao

40

Surgery and Baseplating / Re: nothing but gray

« Last post by kjlawlor on July 31, 2018, 04:40:22 PM »

Hi Daniel,

Thank you so much for the reply. I have reached out to Tad Blair's lab and they have been very helpful regarding the viruses and other information. They did indeed use AAV9.

Hsiao,

I bought the virus from addgene as well, https://www.addgene.org/51085/
So far, the histology shows that the virus expresses, however we have had to dilute it 1:10 but we are still experiencing over-expression so we may need to dilute even further.
We are using the relay lens from inscopix (8.4mm), and the objective lens from Edmund Optics, 5mm Dia. x 12.5mm FL, MgF2 Coated, Achromatic Doublet Lens.
We have not successfully seen any dynamic cells, and I believe that may be either due to the virus or possibly the lens implantation process. We are still troubleshooting to figure out how to improve. Hope your second surgery turns out well!

Kristen