病理核心室 Pathology Core

X-Clarity 送件及收費辦法
X-Clarity Service Notice

• 送件：使用者做好前處理後，將樣本以50ml離心管或Multi-plate盛裝，浸泡在PBS中，送交病理核心。前處理辦法可參考病理核心網頁：https://bit.ly/2PhGpt0；申請單下載：https://bit.ly/2OegVQI
   
• 上機：依送件順序隨到隨上，約需二到三個工作天，視組織大小及數目而定，可能需要更久。
   
• 收費：由於組織大小不一，依實際使用的耗材量計費。價格表如下：

品項	單價	單位
Hydrogel solution		ml
ETC cleaning buffer		hour (電泳時間)
Mounting solution (RI-matching) (optional)*		ml

一顆小鼠全腦澄清化約需 7 ml Hydrogel solution + 6 hour ETC time，所需耗材費用為15 x 7 + 120 x 6 = 825。
 

• 各實驗室可自行購買或配製RI=1.44-1.46之RI-matching solution，於免疫染色後、上機觀察前使用，俾使組織達到完全澄清。如需領用則以上表價格計費。
   

• 澄清完成後的組織會保存在1x PBS中，並以電子郵件通知使用者領回。
   
• 由於電泳液中含有SDS，使用者領回組織後，請更換乾淨1x PBS，於室溫shaker上清洗至少24小時，避免殘留的SDS影響後續免疫染色，或在RI matching solution中形成沉澱物。較大的組織可能需要清洗更久或使用增加細胞通透性的detergent。
   
• 注意：在PBS中組織會變得不透明，在免疫染色後、上機觀察前，浸泡於X-Clarity Mounting Solution或其他RI matching solution，37℃一小時，即可恢復澄清。如果尚未澄清，更換新的X-Clarity Mounting Solution並延長浸潤時間。
   

X-Clarity Mounting Solution會造成螢光訊號降解，在浸泡之後應盡快照相。樣本於X-Clarity Mounting Solution中勿留置超過一星期。           

• Prepare the tissues according to the instructions here: https://bit.ly/2PhGpt0
   

Store the fixed and washed tissues in 50 ml tubes or multi-plates containing 1x PBS. Send the tissues and request form to Pathology Core.
 

Download the service request form here: https://bit.ly/2OegVQI
 

• The fee will be charged according to the actual consumption of reagents. The unit prices are listed below:

Reagent	Unit price	Unit
Hydrogel solution		ml
ETC cleaning buffer		hour (electrophoresis time)
Mounting solution (RI-matching) (optional)*		ml

Example: For clearing one whole mouse brain, it will need 7 ml of hydrogel solution and 6 hours of electrophoresis time. So the fee will be 15 x 7 + 120 x 6 = 825.

* Users can use RI matching solution with a RI of 1.44-1.46 from any manufacturer. Also, users can buy X-Clarity Mounting Solution (Logosbio inc.) from Pathology Core.
 

• Pathology Core will inform users via e-mail when the tissue clearing is done. Cleared samples will be kept in 1x PBS.
   
• Before antibody labeling or imaging, please wash tissues in 1x PBS for at least 24 hours at RT with gentle shaking to remove SDS, which inhibits antibody labeling and will form precipitates in RI matching solutions. For large tissues, prolonged wash time and use of detergents that increase cell permeability are recommended. Following is a washing protocol provided by our user for a whole adult mouse brain:
   

1x PBS + 1% of Triton X-100 for 1-2 days followed by 1x PBS for another 1-2 days
 

• NOTE: Tissues will turn opaque in PBS. To regain transparency, incubate tissues in X-Clarity Mounting Solution or other RI matching solutions for 1 hour at 37℃. Replace with fresh X-Clarity Mounting Solution and incubate for another 1 hour if needed.

ATTENTION: Prolonged exposure to X-Clarity Mounting Solution will lead to decay of the fluorescence signal. Please image samples as soon as possible and DO NOT leave samples in X-Clarity Mounting Solution for more than 1 week.