Whole brain clarity 48hrs ETC

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

Comments

  • 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.
  • bcpsbcps Posts: 1
    Hi Neon,

    Thanks for the posting. Unfortunately I too cannot access the folder
  • NeonNeon Posts: 7
    Sorry, I changed the access level to all users. If not works again, I will post using another drive instead of google.
  • Hi Neon,

    Unfortunately, it still doesn't work.
  • NeonNeon Posts: 7
    I created a blog page. Please find pictures here.
    http://blog.naver.com/scirus/
  • Hi Neon,

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

    Thx
  • NeonNeon Posts: 7
    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.
  • Hi Neon,

    I'm surprised about the slow flow rate. Did you have any bubble forming or was did your solution stay clear?
  • NeonNeon Posts: 7
    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.
  • irinairina Posts: 6
    Hi Neon,
    Really nice results! What temperature did you use for the ETC?
  • mhostmhost Posts: 1
    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!
  • arvidarvid Posts: 9
    Hi Neon, looks really good! Can you post pictures of the chamber?
  • NeonNeon Posts: 7
    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.
  • 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
  • Hi Neon, nice result!
    Have you already taken some fluorescence images of the cleared brain?
  • NeonNeon Posts: 7
    Unfortunately, not successful yet.
  • aysegulaysegul Posts: 31
    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?
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