Your institution must subscribe to JoVE's Biology section to access this content.

Fill out the form below to receive a free trial or learn more about access:


Enter your email below to get your free 10 minute trial to JoVE!

We use/store this info to ensure you have proper access and that your account is secure. We may use this info to send you notifications about your account, your institutional access, and/or other related products. To learn more about our GDPR policies click here.

If you want more info regarding data storage, please contact gdpr@jove.com.




Cite this Article

Copy Citation | Download Citations

Park, J., Cunningham, M. G. Thin Sectioning of Slice Preparations for Immunohistochemistry. J. Vis. Exp. (3), e194, doi:10.3791/194 (2007).

Please note that all translations are automatically generated.

Click here for the english version. For other languages click here.




  1. 准备从磁带华侨城平台的模具。
  2. 与华侨城填充模具。冻结范围内的低温恒温器或通过使用粉碎的干冰。
  3. 删除冻结华侨城平台周围的磁带。
  4. 将冻结夹头和低温恒温器的安装阶段,并锁定在夹头的标记。
  5. 组通过OCT平台,直到表面是平坦的。
  6. 低温恒温器冻结阶段,取出重铺华侨城平台和地点。
  7. 华侨城广场组织样本(以前用30%甘油或蔗糖的PBS冻存)。
  8. 准备冻结列外圈形成华侨城以及列的顶部以上约5毫米投影。
  9. 小心冻结柱表面的中心位置上组织样本,并慢慢加入华侨城,直到充满。
  10. 环绕粉碎干冰冻结列。组织和OCT应完全冻结在20-60秒内。
  11. 准备在温度的增加,外圈可去除样品仍然冻结。
  12. 滑出冻结列侧身的样品,在低温恒温器的地方。
  13. 华侨城华侨城平台和位置的标本(组织下来)申请公司的压力表面放置下降。试样迅速冻结到华侨城平台。
  14. 安全夹头到低温恒温器的安装与对齐标记阶段。
  15. 通过华侨城第肤浅的组织标本。
  16. 解冻安装到玻片薄片和冻结或在室温下储存。
  17. 免疫反应可以在玻片上的组织。
  18. 试剂汇集到幻灯片,可以轻轻地激动,如果光敏感,可能会覆盖。
  19. 以后各阶段的反应很容易进行反相幻灯片浪费插座,吸湿排汗的幻灯片,然后应用在未来试剂。

Subscription Required. Please recommend JoVE to your librarian.



Subscription Required. Please recommend JoVE to your librarian.


Name Company Catalog Number Comments
O. C. T. Ted Pella, Inc. 27050
Glycerol Solution 30% Glycerol Solution in PBS w/v
Sucrose Solution 30% Sucrose Solution in PBS w/v



  1. Scoutern, C. W., O'Connor, R., Cunningham, M. Perfusion fixation of research animals. Microsc. Today. 14, 26-33 (2006).
  2. Cunningham, M. G., Connor, C. M., Zhang, K., Benes, F. M. Diminished serotonergic innervation of adult medial prefrontal cortex after 6-OHDA lesions in the newborn rat. Brain Res. Dev. Brain Res. 157, 124-131 (2005).



  1. Thank-you for posting this video. Your device seems to work well with small tissue, but what do you use for freezing whole adult rat brains? Is the powdered dry ice method sufficient alone to reduce freezing artifact if you are freezing a whole adult brain? Do you have an alternative procedure for these applications?   Much appreciated,

    Posted by: Anonymous
    October 23, 2008 - 4:04 AM
  2. Hi Neil, you actually can get a pretty good freeze with finely powdered dry ice. Just cover and let freeze for 3-5 minutes.  I would recommend, however, immersing the whole brain in isopentane that has been chilled (~30 minutes) in dry ice.  Just leave the brain in isopentane for 30 seconds, otherwise it can distort and fracture.  You will then need to let the (very cold, -80 degrees) brain equilibrate (warm up) to your cryostat or microtome temperature in order for it to cut smoothly.
    Good luck!

    Miles G. Cunningham, MD PhD
    Director, Laboratory for Neural Reconstruction
    Administrative Director, McLean Fellowship in
    Neuropsychiatry and Behavioral Neurology
    Chair, ANPA Programs Committee
    Executive Director, Asniya, Inc.
    MRC 333, McLean Hospital
    Harvard Medical School
    115 Mill Street
    Belmont, MA  0²478
    office:     617.855.²051
    fax:         617.855.3199
    page:      617.855.²000

    Posted by: Anonymous
    October 24, 2008 - 5:18 PM
  3. Thanks for this very helpful video. I'm interested in knowing where you purchased the freezing rod and ring that is used to form the sample mold. Alternatively what metal is the rod made of and is there any significance to the length of the freezing rod used?

    Posted by: Ellen Y.
    April 23, 2010 - 3:57 PM

Post a Question / Comment / Request

You must be signed in to post a comment. Please or create an account.

Usage Statistics