Calcium imaging in Multiphoton systems

[Carlos Sanchez-8]

Hello, good afternoon.

 

- From your experience, which are the best calcium dyes to be used in Multiphoton systems? Calcium green, perhaps?

 

- I supposed that excitation ratio dyes like Fura-2 are not recommended but has anybody used it with multiphoton excitation? Is it worthy?

 

Thank you very much in advance for your help.

Regards,

 

Carlos Sánchez Martín

Servicio de Microscopía Optica y Confocal (Lab.310)

Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)

C/Nicolás Cabrera,1

Universidad Autónoma de Madrid. Cantoblanco.

28049. Madrid

Telf: 91 196 4613/4643

Fax: [phone]

E-mail: [hidden email]

http://www.cbm.uam.es/confocal

 

[Mark Cannell]

Indo-1 is good.

See
Jones, K.T., Soeller, C. & Cannell, M.B. (1998) The passage of Ca2+ and
fluorescent markers between the sperm and egg after fusion in the mouse.
Development 125, 4627-4635.

(shameless self promotion I know ;-)

Cheerrs



Carlos Sanchez wrote:

>
> Hello, good afternoon.
>
>  
>
> - From your experience, which are the best calcium dyes to be used in
> Multiphoton systems? Calcium green, perhaps?
>
>  
>
> - I supposed that excitation ratio dyes like Fura-2 are not
> recommended but has anybody used it with multiphoton excitation? Is it
> worthy?
>
>  
>
> Thank you very much in advance for your help.
>
> Regards,
>
>  
>
> Carlos Sánchez Martín
>
> Servicio de Microscopía Optica y Confocal (Lab.310)
>
> Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)
>
> C/Nicolás Cabrera,1
>
> Universidad Autónoma de Madrid. Cantoblanco.
>
> 28049. Madrid
>
> Telf: 91 196 4613/4643
>
> Fax: [phone]
>
> E-mail: [hidden email]
>
> http://www.cbm.uam.es/confocal
>
>  
>

[Ignatius, Mike-2]

Dr. Martin,

 

I can send a recent 4 page brief (or link to the online version) on 2-photon compatible dyes for neuronal physiology.  (It was the cover story in our BioProbes 60 issue last fall.) 

 

Oregon Green Bapta 1 and 2 and the fluo series are popular for Multi-bolus labeling as they are cell permeant and non-ratiometric.  For intracellular injection there are many choices, depends on what calcium range you are monitoring, whether you are seeking absolute concentrations or just monitoring deltas.  This and more is covered in our review.

 

Kind Regards,

 

Mike Ignatius,

 

Molecular Probes/Lifetech

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Carlos Sanchez
Sent: Friday, February 19, 2010 7:49 AM
To: [hidden email]
Subject: Calcium imaging in Multiphoton systems

 

Hello, good afternoon.

 

- From your experience, which are the best calcium dyes to be used in Multiphoton systems? Calcium green, perhaps?

 

- I supposed that excitation ratio dyes like Fura-2 are not recommended but has anybody used it with multiphoton excitation? Is it worthy?

 

Thank you very much in advance for your help.

Regards,

 

Carlos Sánchez Martín

Servicio de Microscopía Optica y Confocal (Lab.310)

Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)

C/Nicolás Cabrera,1

Universidad Autónoma de Madrid. Cantoblanco.

28049. Madrid

Telf: 91 196 4613/4643

Fax: [phone]

E-mail: [hidden email]

http://www.cbm.uam.es/confocal

 

[Roland Nitschke]

Hello,
we have used fura-2 with single wavelength 2-P excitation here:
Simultaneous Measurements of Cytosolic and Mitochondrial Ca2+ Transients in HT29 Cells*
Sigrid Ricken, Jens Leipziger, Rainer Greger, and Roland Nitschke‡
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 273, No. 52, Issue of December 25, pp. 34961–34969, 1998

If you have access to a double 2-P system also excitation ratioing should be possible using app. 690 and 780 nm.

Regards

Roland


Am 19.02.2010 16:49, schrieb Carlos Sanchez:

Hello, good afternoon.

 

- >From your experience, which are the best calcium dyes to be used in Multiphoton systems? Calcium green, perhaps?

 

- I supposed that excitation ratio dyes like Fura-2 are not recommended but has anybody used it with multiphoton excitation? Is it worthy?

 

Thank you very much in advance for your help.

Regards,

 

Carlos Sánchez Martín

Servicio de Microscopía Optica y Confocal (Lab.310)

Centro de Biología Molecular "Severo Ochoa" (CSIC-UAM)

C/Nicolás Cabrera,1

Universidad Autónoma de Madrid. Cantoblanco.

28049. Madrid

Telf: 91 196 4613/4643

Fax: [phone]

E-mail: [hidden email]

http://www.cbm.uam.es/confocal