>Dear Colleagues,
>I would like to be able to do ratio [Ca] measurements in living cells using
>a Bio-Rad MRC 600 c-focal scope equipped with an Argon-Krypton laser.
>I know that it should be possible with dyes such as Calcium Green ratioed
>with e.g., a Texas Red-conjugated dextran.
>However, loading cells with a Texas Red-dextran involves !injection, which
>I want to avoid.
>Any ideas?
>Thank you very much for your time & consideration.
>Michal Opas
>Anatomy & Cell Biology
>U of Toronto
>
>
 
After attending the Bio-Rad training conference last September, we received
some information regarding Ion imaging. I think I understand what you are
asking but I'm not totally sure. I will give you what info that I have and
if this doesn't help, I suggest that you call Bio-Rad and ask them to send
you some info.
 
Indicators for Laser-excited Ion imaging
 
Ion           Excitation Ratioing  Emission Ratioing    Single Wavelength
 
Calcium       Fura Red 488 442     Indo-1     351       Calcium Green  488
                                                        Calcium Orange 543
                                                        fluo-3         488
 
 
For preparation they suggest:
 
1. Grow cells or do the appropriate preparation.
 
2. Load cells with indicator dye by one of:
 
 
        1) Soaking in ester form of the dye (wait for hydrolysis)
        2) Microinjection of free acid or dextran conjugate
        3) "Scrape" loading, electroporation, ect.
 
I hope that this helps!
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+ Brett Schroeder                            8701 Watertown Plank Road +
+ Dept. of Cellular Biology & Anatomy     Milwaukee, WI 53226-4801 USA +
+ Medical College of Wisconsin                          (414) 456-8504 +
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