Whole brain clarity 48hrs ETC

Neon

With a modified chamber design and electric condition, I got the fast ETC result. 1A, 30V used for 48hrs.

https://drive.google.com/#folders/0B5k-H33OqLCjdUduNnJ4dV9wcUE

DominicJG

Hi Neon,

Happy New Year.

It sounds interesting. Unfortunately, access to the linked folder was denied. Could you give permission to your linked folder?

Thank you.

bcps

Hi Neon,

Thanks for the posting. Unfortunately I too cannot access the folder

Neon

Sorry, I changed the access level to all users. If not works again, I will post using another drive instead of google.

DominicJG

Hi Neon,

Unfortunately, it still doesn't work.

Neon

I created a blog page. Please find pictures here.
http://blog.naver.com/scirus/

DominicJG

Hi Neon,

Thanks for setting up the blog. Your results look very nice. I was wondering how you modified your chamber?

Thx

Neon

I cooled chamber directly instead of using water circulator. Because I didn't tried DIY version, don't know yet what made the difference. Minimized turbulance might be one. I used peristaltic pump to exchange buffer. Quite slow flow rate, 20-30 ml/min, instead of L/min level.

DominicJG

Hi Neon,

I'm surprised about the slow flow rate. Did you have any bubble forming or was did your solution stay clear?

Neon

Hi Dominic,
I also noticed the bubble forming was increased with the decreased flow rate. But it seemed not a critical issue with my system. With the mentioned flow rate, the bubble layer was stayed on top of the chamber (about 10% of total chamber height), and the tissue was located at the center position.

irina

Hi Neon,
Really nice results! What temperature did you use for the ETC?

mhost

Hi Neon,
When you modified your chamber you just added a pump and slowed the flow rate? We are very interested in your protocol and design. We are having a lot of problems with bubble formation and predict it is having a negative impact on our sample but you believe the bubble formation in your design does not effect the sample? Any additional information about your design would be greatly appreciated. Thanks!

arvid

Hi Neon, looks really good! Can you post pictures of the chamber?

Neon

Hi irina, mhost,
I used the temperature 37 degree C. Because the slow flow rate will make a temperature difference b/w chamber and buffer, and it did, I directly cooled the chamber. Also I measured the temperature of the chamber instead of buffer and tried to maintain a constant temperature inside of the chamber. Because I didn't do much experiment using different condition, I do not have many trouble shooting at this moment. Not sure why the bubble makes some problems at other system. I would be pleased to share more data after doing more experiments.

Shashank20

Hi Neon, Your results are very encouraging. Would it be possible to upload the picture of the modified chamber that you used ? I do not get any success even after 4 days of ETC. What was the temperature of the chamber when you ran ETC ? Thank you

derby_155

Hi Neon, nice result!
Have you already taken some fluorescence images of the cleared brain?

Neon

Unfortunately, not successful yet.

aysegul

Hi Neon,

I cannot access to your blog page. Would you please share your experiment photos? Also i used rat brain but it tarnished during ETC. Our ETC conditions; 37 degree C, propeller (for buffer circulation), 0.4 A, 24 V. After 8 hour later ,discoloration started. Would it be possible to upload the picture of your modified chamber?