stable CFP and YFP in Arabidopsis?

[Christian-103]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I was wondering if anyone knew where I might find some stable CFP and YFP Arabidopsis seed?  The localization does not matter, I only need to practice with the two proteins in plant material.  I have users coming with "YFP" which is GFP, and a few are starting to use CFP and I'm a bit leery of CFP/GFP crosstalk.  Thusly I have need for practice.

Thanks.

Christian

[Hanna_SN]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear Christian,

There is a publication by Yuda Fang and David L. Spector "Centromere
Positioning and Dynamics in Living Arabidopsis Plants" Mol Biol Cell (2005)
Vol. 16, 5710–5718, in which the authors use HTB1-CFP/HTR12-YFP transformed
Arabidopsis plants. Maybe try to contact the authors for the seeds.

All the best,
Hanna

2012/1/6 Christian <[hidden email]>

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I was wondering if anyone knew where I might find some stable CFP and YFP
> Arabidopsis seed?  The localization does not matter, I only need to
> practice with the two proteins in plant material.  I have users coming with
> "YFP" which is GFP, and a few are starting to use CFP and I'm a bit leery
> of CFP/GFP crosstalk.  Thusly I have need for practice.
>
> Thanks.
>
> Christian
>

--
Department of Plant Anatomy and Cytology
Faculty of Biology and Environmental Protection
University of Silesia
Jagiellonska Str. 28
40-032 Katowice
Poland

[Rosemary.White]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Christian,

If you do sequential scanning you can easily separate CFP, GFP and YFP in
a single plant cell, and have a channel for chlorophyll fluorescence as
well (same pass as GFP).

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E [hidden email]


On 7/01/12 8:33 PM, "Hanna Sas Nowosielska" <[hidden email]>
wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Dear Christian,
>
>There is a publication by Yuda Fang and David L. Spector "Centromere
>Positioning and Dynamics in Living Arabidopsis Plants" Mol Biol Cell
>(2005)
>Vol. 16, 5710­5718, in which the authors use HTB1-CFP/HTR12-YFP
>transformed
>Arabidopsis plants. Maybe try to contact the authors for the seeds.
>
>All the best,
>Hanna
>
>2012/1/6 Christian <[hidden email]>
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> I was wondering if anyone knew where I might find some stable CFP and
>>YFP
>> Arabidopsis seed?  The localization does not matter, I only need to
>> practice with the two proteins in plant material.  I have users coming
>>with
>> "YFP" which is GFP, and a few are starting to use CFP and I'm a bit
>>leery
>> of CFP/GFP crosstalk.  Thusly I have need for practice.
>>
>> Thanks.
>>
>> Christian
>>
>
>
>
>--
>Department of Plant Anatomy and Cytology
>Faculty of Biology and Environmental Protection
>University of Silesia
>Jagiellonska Str. 28
>40-032 Katowice
>Poland

[Littlejohn, George]

Hi Christian, Rosemary, List,

Absolutely. We use Yellow CaMeleon FRET-based Calcium sensors in Arabidopsis. This relies on spectral separation of CFP and YFP. Feel free to contact me off list if I can advise on settings or help with plant lines.

All the best,
George


Dr. George Littlejohn
University of Exeter
College of Life and Environmental Sciences
Biosciences
Geoffrey Pope Building
Mezzanine Lab
Stocker Road
Exeter
United Kingdom
EX4 4QD

******************************
Tel:  [phone] (Lab.)
       [phone] (Office)
Fax:  [phone]
E-mail:  [hidden email]
******************************
http://www.illuminatedcell.com/improved-imaging.html


On 8 Jan 2012, at 22:50, "Rosemary White" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Christian,
>
> If you do sequential scanning you can easily separate CFP, GFP and YFP in
> a single plant cell, and have a channel for chlorophyll fluorescence as
> well (same pass as GFP).
>
> cheers,
> Rosemary
>
> Dr Rosemary White
> CSIRO Plant Industry
> GPO Box 1600
> Canberra, ACT 2601
> Australia
>
> T 61 2 6246 5475
> F 61 2 6246 5334
> E [hidden email]
>
>
> On 7/01/12 8:33 PM, "Hanna Sas Nowosielska" <[hidden email]>
> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Dear Christian,
>>
>> There is a publication by Yuda Fang and David L. Spector "Centromere
>> Positioning and Dynamics in Living Arabidopsis Plants" Mol Biol Cell
>> (2005)
>> Vol. 16, 5710­5718, in which the authors use HTB1-CFP/HTR12-YFP
>> transformed
>> Arabidopsis plants. Maybe try to contact the authors for the seeds.
>>
>> All the best,
>> Hanna
>>
>> 2012/1/6 Christian <[hidden email]>
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> I was wondering if anyone knew where I might find some stable CFP and
>>> YFP
>>> Arabidopsis seed?  The localization does not matter, I only need to
>>> practice with the two proteins in plant material.  I have users coming
>>> with
>>> "YFP" which is GFP, and a few are starting to use CFP and I'm a bit
>>> leery
>>> of CFP/GFP crosstalk.  Thusly I have need for practice.
>>>
>>> Thanks.
>>>
>>> Christian
>>>
>>
>>
>>
>> --
>> Department of Plant Anatomy and Cytology
>> Faculty of Biology and Environmental Protection
>> University of Silesia
>> Jagiellonska Str. 28
>> 40-032 Katowice
>> Poland

[Christian-103]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Thank you all for the input.  My primary concern is practicing with known samples.  The autofluorescence in plants of course shows up strongly in the "DAPI" range, weakly in the "GFP" range and of course very strongly in the "CY 5" range.  With new fluorescent proteins slowly trickling in, I am working on getting more comfortable with them.  I miss the good ol' days when I worked in simple things from animal tissues!  I'm kidding, I love the challenge of plant materials.

Christian



--- On Sun, 1/8/12, Littlejohn, George <[hidden email]> wrote:

From: Littlejohn, George <[hidden email]>
Subject: Re: stable CFP and YFP in Arabidopsis?
To: [hidden email]
Date: Sunday, January 8, 2012, 5:52 PM

Hi Christian, Rosemary, List,

Absolutely. We use Yellow CaMeleon FRET-based Calcium sensors in Arabidopsis. This relies on spectral separation of CFP and YFP. Feel free to contact me off list if I can advise on settings or help with plant lines.

All the best,
George


Dr. George Littlejohn
University of Exeter
College of Life and Environmental Sciences
Biosciences
Geoffrey Pope Building
Mezzanine Lab
Stocker Road
Exeter
United Kingdom
EX4 4QD

******************************
Tel:  [phone] (Lab.)
       [phone] (Office)
Fax:  [phone]
E-mail:  [hidden email]
******************************
http://www.illuminatedcell.com/improved-imaging.html



On 8 Jan 2012, at 22:50, "Rosemary White" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

> *****
>
> Hi Christian,
>
> If you do sequential scanning you can easily separate CFP, GFP and YFP in
> a single plant cell, and have a channel for chlorophyll fluorescence as
> well (same pass as GFP).
>
> cheers,
> Rosemary
>
> Dr Rosemary White
> CSIRO Plant Industry
> GPO Box 1600
> Canberra, ACT 2601
> Australia
>
> T 61 2 6246 5475
> F 61 2 6246 5334
> E [hidden email]
>
>
> On 7/01/12 8:33 PM, "Hanna Sas Nowosielska" <[hidden email]>
> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

>> *****
>>
>> Dear Christian,
>>
>> There is a publication by Yuda Fang and David L. Spector "Centromere
>> Positioning and Dynamics in Living Arabidopsis Plants" Mol Biol Cell
>> (2005)
>> Vol. 16, 5710­5718, in which the authors use HTB1-CFP/HTR12-YFP
>> transformed
>> Arabidopsis plants. Maybe try to contact the authors for the seeds.
>>
>> All the best,
>> Hanna
>>
>> 2012/1/6 Christian <[hidden email]>
>>
>>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy

>>> *****
>>>
>>> I was wondering if anyone knew where I might find some stable CFP and
>>> YFP
>>> Arabidopsis seed?  The localization does not matter, I only need to
>>> practice with the two proteins in plant material.  I have users coming
>>> with
>>> "YFP" which is GFP, and a few are starting to use CFP and I'm a bit
>>> leery
>>> of CFP/GFP crosstalk.  Thusly I have need for practice.
>>>
>>> Thanks.
>>>
>>> Christian
>>>
>>
>>
>>
>> --
>> Department of Plant Anatomy and Cytology
>> Faculty of Biology and Environmental Protection
>> University of Silesia
>> Jagiellonska Str. 28
>> 40-032 Katowice
>> Poland