Re: Re: RE: [Histonet] Anterograde tracing


Hi all again.
Just wonder anyone tried WGA-HRP?

it seems that WGA-hRP has a good diffusion ability,, making it ideal for local pressure injection (quite simple method), which PHA-L does bad.
and you can detect it well.

anyone have worked with this?




发件人: tf 
发送时间: 2008-07-12  00:50:29 
收件人: Rachel, Rivka (NIH/NEI) [E]; histonet 
主题: Re: RE: [Histonet] Anterograde tracing 
Dear Rachel:
Surely I have considered that.
I am actually working on regeneration.
Also, dyes are versatile and go everywhere as long as membrane permits. It does not have the restricted labeling of of one batch of projection and may diffuse a lot when tracing the transection site. Another problem you have mentioned is that DiI does not diffuse well in adult tissue, and thus used more in developmental studies.
thanks a lot
发件人: Rachel, Rivka (NIH/NEI) [E] 
发送时间: 2008-07-12  00:00:49 
收件人:; histonet 
主题: RE: [Histonet] Anterograde tracing 

What type of tissue are you trying to trace?  If fixed, immature rodent tissue, have you considered one of the fluorescent tracers such as DiI, DiO, DiD, etc. available through Molecular Probes (Invitrogen)?  DiI (red fluorescence) is the brightest.  These dyes don't work well in adult tissue, however.
Rivka A. Rachel, MD, PhD
Staff Scientist, National Eye Institute
Neurobiology-Neurodegeneration and Repair Laboratory
Tel: 301 443-4906
-----Original Message-----
From: tf []
Sent: Fri 7/11/2008 11:41 AM
To: histonet
Subject: [Histonet] Anterograde tracing
I just want to discuss the best anterograde tracer.
BDA, PHA-L, CTB-FITC, CTB-Rhodamine, WGA-HRP, HRP, and viral vectors all could be anterograde tracer.
However, BDA is diffusible like all fluroscent dyes, does not have the necessary direction.
CTB-conjugated sereis have limited diffusion ability when injected with pressure, just like PHA-L, thus are not good in labeling a larger group of neurons when single rather multiple injections is required.
WGA-HRP & HRP both antero & retrogradely transported, HRP & virus even transynaptically.
So, any one has other ideas? Such as shorten the period after injection to reduce the diffusion, or using immunodetection to visualize the CTB-FITC, for example?
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