intra-vital nuclear marker

[Kurt Anderson-3]

hi ho,

i am looking for an injectable red-fluorescent nuclear marker comparable to hoechst 33342 for intra-vital mouse imaging.

hoechst is highly specific, and easy to excite at 850 nm, but the imaging penetration depth is severely limited by scattering of blue light. we can see GFP much deeper and with better clarity (resolution and S:N).

it has to be injectable for easy combination with our various GFP mouse lines, so RFP-Histone 2B is out.

DRAQ5 works great on cells in culture under 1P and 2P excitation. but i have not been able to visualise it following tail vein injection using a protocol which works fine for hoechst (inject, wait an hour, image). has anyone used DRAQ5 for intra-vital imaging?

SYTO59 works on cells in culture and in tissue following tail vein injection under 1P and 2P excitation. but it lacks the specificity of hoechst; i.e. its a general nucleic acid label so you get a lot of cytoplasmic signal and nuclear boundaries are not as clearly defined.

TOPRO, YOYO, GaGa, GooGoo... help!

anyone got a suggestion?

thanks,

kurt

.

Dr. Kurt I. Anderson

Actin Dynamics and Beatson Advanced Imaging Resource (BAIR)

The Beatson Institute for Cancer Research

Garscube Estate, Switchback Road, Glasgow  G61 1BD

 

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[Glen MacDonald-2]

The cyanine nuclear markers, To-Pro, Yo-Yo, etc are membrane impermeant.  They will enter only cells expressing channels  for divalent cations, such as P2X7 purinergic channels, and a few other channels not yet characterized.  

Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]
On Feb 23, 2010, at 8:37 AM, Kurt Anderson wrote:

> hi ho,
>
> i am looking for an injectable red-fluorescent nuclear marker comparable to hoechst 33342 for intra-vital mouse imaging.
>
> hoechst is highly specific, and easy to excite at 850 nm, but the imaging penetration depth is severely limited by scattering of blue light. we can see GFP much deeper and with better clarity (resolution and S:N).
>
> it has to be injectable for easy combination with our various GFP mouse lines, so RFP-Histone 2B is out.
>
> DRAQ5 works great on cells in culture under 1P and 2P excitation. but i have not been able to visualise it following tail vein injection using a protocol which works fine for hoechst (inject, wait an hour, image). has anyone used DRAQ5 for intra-vital imaging?
>
> SYTO59 works on cells in culture and in tissue following tail vein injection under 1P and 2P excitation. but it lacks the specificity of hoechst; i.e. its a general nucleic acid label so you get a lot of cytoplasmic signal and nuclear boundaries are not as clearly defined.
>
> TOPRO, YOYO, GaGa, GooGoo... help!
>
> anyone got a suggestion?
>
> thanks,
>
> kurt
>
> .
>
>
>
>
> Dr. Kurt I. Anderson
> Actin Dynamics and Beatson Advanced Imaging Resource (BAIR)
> The Beatson Institute for Cancer Research
> Garscube Estate, Switchback Road, Glasgow  G61 1BD
>  
> ( Direct Line [phone]
> ( Fax                [phone]
> * E-mail          [hidden email]
>  
> A company limited by guarantee; Registered in Scotland No 84170; A Registered Scottish Charity No SC006106
>  
> The information contained in this communication may be privileged and confidential and is intended for the exclusive use of the addressee designated above. If you are not the addressee, any disclosure, reproduction, distribution or other dissemination or use of this communication is strictly prohibited. If you have received this transmission in error please would you be kind enough to inform us.
>  
>