Ca imaging in brain slices

[Barry O'Brien]

Query posted on behalf of the colleague listed below.

>This is not a confocal question, but more a methodological question.
>We want to image seizure-related calcium
>fluctuations within rodent cerebral cortical tissue (400um coronal
>slices), using the Fura-2AM dye. Does anyone have experience using this
>dye to capture calcium fluctuations in cortical slice tissue? In
>particular, is there an optimal dye loading technique for this tissue
>which will maximise the calcium signal.

>We would be grateful for any help/suggestions.
>
>We are using a Nikon AZ100 with epifluorescence. Camera is a QuantEM 512SC
>electron multiplying CCD. We have Fura-2 filter set (Semrock
>Brightline); excitation 340nm/380nm; emission 510nm; dichroic
>344-404nm reflection band/415-570nm transmission band.
>
>
>--------------------------------------------------------------
>Dr Logan Voss
>Research Fellow
>Waikato Clinical School
>Hamilton
>New Zealand
>phone: [phone] ext8406
>mobile: [phone]

"Logan Voss" <[hidden email]>


Dr Barry O'Brien
Dept of Biological Sciences,
University of Waikato
Private Bag 3105
HAMILTON
New Zealand

Fax [phone]
Phone [phone]

[regan m]

Hi,

 

In the past i have used Fura 2AM dye in neuronal cell lines

The thing is dye uptake can be an issue and if your are planning to setup the chamber where you culture the cells right on the stage then it shud help u circumvent certain problems like cells rounding up ETC

 

I used 2uM concentration of dye .. Its not toxic Incubate the cells with dye for about 2 hrs before imaging  

I think it shud work fine let me know if u need some specific help.

 

Regards,

Johnson.

On Tue, Feb 17, 2009 at 4:44 AM, Barry O'Brien <[hidden email]> wrote:

Query posted on behalf of the colleague listed below.

This is not a confocal question, but more a methodological question. We want to image seizure-related calcium
fluctuations within rodent cerebral cortical tissue (400um coronal
slices), using the Fura-2AM dye. Does anyone have experience using this
dye to capture calcium fluctuations in cortical slice tissue? In
particular, is there an optimal dye loading technique for this tissue
which will maximise the calcium signal.

We would be grateful for any help/suggestions.

We are using a Nikon AZ100 with epifluorescence. Camera is a QuantEM 512SC
electron multiplying CCD. We have Fura-2 filter set (Semrock
Brightline); excitation 340nm/380nm; emission 510nm; dichroic
344-404nm reflection band/415-570nm transmission band.


--------------------------------------------------------------
Dr Logan Voss
Research Fellow
Waikato Clinical School
Hamilton
New Zealand
phone: [phone] ext8406
mobile: [phone]

"Logan Voss" <[hidden email]>


Dr Barry O'Brien
Dept of Biological Sciences,
University of Waikato
Private Bag 3105
HAMILTON
New Zealand

Fax [phone]
Phone [phone]