Nabble - Bio.net - Fluorpro

LED for fluorescence

2009-12-09T12:38:13Z

Hello all, and seasonal greetings!

I am considering purchasing a fluorescence scope for my lab. this
is a work-horse scope - for trouble shooting, getting incubation and
fixation times right etc. The pretty pictures will be taken at a core
facility, with a fancy schmanzy scope.

The fluors I use are DAPI, FITC and/or GFP, TRITC and/or rhodamine.

I want to avoid yucky mercury or metal halide lamps wot might get left
on and blow up or at the least cost a fortune to replace. I am
considering a bottom of the line research grade scope (I think called
Aksioscop A1) from Zeiss with their clever LED illumination source.
This source includes separate LEDs for each excitation frequency.

Anybody out there have experience with this? It aint cheap but i have
the money now and probably never will again so carpe diem etc. (or
perhaps caveat emptor???).

Hugely greatful for any feedback,

Thanks,

Sandra

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Only few days left: Register now for our SPM & Optical Tweezers Symposium

2009-09-25T08:03:30Z

Dear Colleague,

We would like to remind you of the Joint Meeting on the Application of SPM & Optical Tweezers for Life Sciences, taking place in Berlin, October 14 – 15 this year.

This symposium is a wonderful opportunity to hear eighteen of the world's leading scientists of SPM and Optical Tweezers in life sciences applications as well as to present your own work as a poster.

Please visit our website www.nanobioviews.net/registration.html for registration and www.nanobioviews.net/talks.html for detailed information.

We look forward to welcoming you and your colleagues to Berlin.

Kind regards,

Petra Dammermann

--
Petra Dammermann
NanoBioVIEWS

Tel: +49 30 5331 12070
Fax: +49 30 5331 22555
mail: info@...
web: www.nanobioviews.net

Aufsichtsrat: Dr. Franz-Ferdinand von Falkenhausen (Vorsitzender)
Vorstand: Torsten Jähnke, Frank Pelzer, Jörn Kamps, René Grünberg

JPKinstruments Aktiengesellschaft
Amtsgericht Berlin-Charlottenburg
HRB 75513

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Early registration ending next Monday - Joint SPM & Optical Tweezers Meeting in Berlin

2009-08-26T09:44:03Z

Dear Colleague,

we would like to remind you of the Joint Meeting on the Application of SPM & Optical Tweezers for Life Sciences, taking place in Berlin, October 14 – 15 this year.

This symposium is a wonderful opportunity to hear eighteen of the world's leading scientists of SPM and Optical Tweezers in life sciences applications as well as to present your own work as a poster.

Please visit our website www.nanobioviews.net/registration.html for registration and www.nanobioviews.net/talks.html for detailed information.

We look forward to welcoming you and your colleagues to Berlin.

Kind regards,

Petra Dammermann

________________
Petra Dammermann
NanoBioVIEWS

JPK Instruments AG
Bouchéstrasse 12
12435 Berlin

tel: +49 30 5331 12070
fax: +49 30 5331 22555
mail: info@...
web: www.nanobioviews.net

Aufsichtsrat: Dr. Franz-Ferdinand von Falkenhausen (Vorsitzender)
Vorstand: Torsten Jähnke, Frank Pelzer, Jörn Kamps, René Grünberg

JPKinstruments Aktiengesellschaft
Amtsgericht Berlin-Charlottenburg
HRB 75513

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Joint Meeting on the Application of SPM & Optical Tweezers for Life Sciences

2009-07-09T06:49:41Z

Dear Colleague,

On behalf of NanoBioVIEWS™, JPK Instruments invites you to attend the 2009 Symposia on the application of Scanning Probe Microscopy (SPM) and optical tweezers for the Life Sciences. The meeting will run for two days in Berlin on October 14-15^th. Delegates are welcome to attend either the full event or focused SPM (October 14) or Optical Tweezers (October 15) sessions.

Oral presentations from leading research groups in Europe and the USA will be complemented with poster sessions providing an open forum for delegates to discuss their latest research. A wide range of topics will be covered including imaging, interactions and dynamics in Life Sciences from single molecules to cellular systems.

For further details, a full program, and registration please visit: www.nanobioviews.net.

We look forward to welcoming you and your colleagues to Berlin in October.

Kind regards

Petra Dammermann

_________________
Petra Dammermann
NanoBioVIEWS™

Bouchéstrasse 12
12435 Berlin

tel: +49 30 5331 12070
fax: +49 30 5331 22555
mail: info@...
web: www.nanobioviews.net

JPKinstruments Aktiengesellschaft
Handelsregister: Amtsgericht Berlin-Charlottenburg
Registernummer: HRB 75513
Vorstand: Torsten Jähnke, Frank Pelzer, Jörn Kamps, René Grünberg
Aufsichtsrat: Dr. Thomas van Aubel (Vorsitzender)

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GFP users. please guide

2009-06-09T16:03:35Z

Hey,

My name is Maria Teresa George a student doing my Masters at the Claremont Colleges in California.

For one of my research I need to determine the number of labs around the world (or atleast in the US ) or the number of users who are doin GFP cell based research. Can you give me any inputs on how I can arrive at the number. Any form of inputs would be really helpful.

Thank you so much for you help

Sincerely,

Maria
--
Maria Teresa George
Class of 2010
Keck Graduate Institute
Claremont Colleges
909-702-0250
maria_george@...

--
Maria Teresa George
Class of 2010
Keck Graduate Institute
Claremont Colleges
909-702-0250
maria_george@...
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Quenching Fluorescent Proteins

2008-11-21T02:55:09Z

Hello,

I'm looking for a way to reliably inactivate fluorescent proteins
(EGFP and dsRed) without harming epitopes for immuno.

A little background: I'm electroporating inducible constructs into
the chick embryo. We use dsRed as the electroporation control, and
EGFP as a marker for activation of our doxicyclin-inducible construct.
We want to use immunofluorescence to on these samples. We currently
use 4% formalin to fix, washes with PBS+tween, usually ethanol
dehydration for storage, and sucrose-agar embedding for sectioning,
and the fluorescence still comes up strong. Obviously, imaging with
epifluorescence with a green and red channel, we can't add another
fluorophore.

Best case would be a way to specifically kill the RFP leaving the
green intact, although as long as we can recover the GFP with an
antibody, this isn't a problem. Also looking for something that won't
damage our epitopes to badly!

Thanks,
Dave

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DsRed stable transfection doesn't Fluoresce?

2008-11-03T07:01:28Z

Hi,

I am have trouble with getting stable expression of DsRed in HEK
cells. I am using the <a href="http://www.ncbi.nlm.nih.gov/pubmed/
11730010">
pStoplight vector</a> which express DsRed until it is cut out by Cre
Recombinase, and then GFP is expressed. I have cloned this into the <a
href="http://products.invitrogen.com/ivgn/en/US/adirect/invitrogen?
cmd=catProductDetail&entryPoint=adirect&productID=K601001&messageType=catProductDetail&showAddButton=true">Flip-
in kit</a> from Invitrogen to generate a stable cell line using flp
recombinase to integrate the vector.

It looks good for a while, but then, even though I have cells
surviving the selection agent, after a week or so, I dont have any
more red fluorescent cells!

So, I have no idea whats happening. It seems to me that either: 1) the
transient expression of flp recombinase is somehow acting on all the
stoplight transfected cells, somehow removing the gene... unlikely. 2)
DsRed expression is somehow being driven down by the cell? 3) DsRed is
somehow toxic and kills the stable cells, but then why does anything
survive!?

Anyway, any suggestions or advise would be very much appreciated!
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Re: Advice on mammalian cells

2008-10-14T01:24:18Z

Hi,

in my experience, 3T3 mouse fibroblasts are easy to culture, easy to
transfect and give nice pictures. They should be easy to obtain, maybe
you even know a lab doing cell culture, they might have them.

Wo
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Advice on mammalian cells

2008-10-13T11:18:04Z

We would like to do YFP localization and BiFC assays in a mammalian cell line. Does anyone have any recommendations on with cell lines to buy that are easy to transfect and image using confocal microscopy?

Thank you in advance for your time

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notisblokk. http://no.mail.yahoo.com

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SPM in Life Sciences Symposium in Berlin

2008-09-24T04:59:12Z

Dear Colleague,

We just would like to remind you of the 7th International Symposium on SPM in Life Sciences, taking place in Berlin, October 8-9 this year.

This symposium is a wonderful opportunity to hear fifteen of the world's leading scientists of SPM in life sciences applications as well as to present your own work as a poster. The deadline for abstracts has been extended to September 30.

Please visit our website www.nanobioviews.net for detailed information.

We look forward to welcoming you and your colleagues to Berlin.

Kind regards,

Petra Dammermann

________________
Petra Dammermann
NanoBioVIEWS™

Bouchéstrasse 12
12435 Berlin

tel: +49 30 5331 12070
fax: +49 30 5331 22555
mail: info@...
web: www.nanobioviews.net

JPKinstruments Aktiengesellschaft
Handelsregister: Amtsgericht Berlin-Charlottenburg
Registernummer: HRB 75513
Vorstand: Torsten Jähnke, Frank Pelzer, Jörn Kamps, René Grünberg
Aufsichtsrat: Dr. Thomas van Aubel (Vorsitzender)

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7th International Symposium on SPM in the Life Sciences

2008-09-02T05:01:07Z

Dear Colleagues,

as previously announced, on behalf of NanoBioViews™, JPK Instruments will organise the 7^th International Symposium on SPM in the Life Sciences in Berlin on October 8-9^th.

Registration is now open!

Please visit our website www.nanobioviews.net/spm-symposium.html for registration and detailed information.

Due to tense hotel situation in Berlin we strongly recommend to book your accommodation as soon as possible. We have reserved a number of rooms at hotels, which can be accessed through our website:

www.nanobioviews.net/spm-hotellist08.html

We look forward to welcoming you and your colleagues to Berlin in October.

Kind regards,

Petra Dammermann

________________

Petra Dammermann

NanoBioVIEWS™

Bouchéstrasse 12

12435 Berlin

tel: +49 30 5331 12070

fax: +49 30 5331 22555

mail: info@...

web: www.nanobioviews.net

JPKinstruments Aktiengesellschaft

Handelsregister: Amtsgericht Berlin-Charlottenburg
Registernummer: HRB 75513
Vorstand: Torsten Jähnke, Frank Pelzer, Jörn Kamps, René Grünberg
Aufsichtsrat: Dr. Thomas van Aubel (Vorsitzender)

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Questions about protein quantification

2008-05-26T06:13:13Z

Hi,

I've recently transferred from academia into industry and have stumbled
onto a problem.

I need a tag that can be used N-terminally to quantify expression of its
fusion partner - ideally in vivo. I was thinking about GFP, or one of
its derivates, but I think this protein might be patented. I've tried
searching for relevant patents without success. Can anyone confirm that
I need a license for the use of GFP (or any of its derivates) for
industrial gene expression quantification?

What alternatives are there? Can luciferase be used in vivo (will
luciferin enter the cells)? Is its use protected?

I'd rather not use antibodies since this is expresive, cumbersome and
must be done in vitro.

Kind regards,
Trond Erik Vee Aune
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This week's microscopy webinar schedule

2008-01-06T14:39:37Z

You are invited to attend our upcoming live, interactive, web-based
instructional microscopy seminars.

Pre-registration is required and connection lines are limited so
reserve yours now. There is no charge to participate.

See further descriptions and pre-register at:
http://www.magbiosystems.com/education

Coming up this week (January 7 - 11):

==========================================================
"Quantitative Image and Data Acquisition for Fluorescent Specimens"
Advice from a Facility Director

Presented by Brian Matsumoto, Ph.D., University of California, Santa
Barbara

Monday, 07-January at 1:30 PM (New York time)

==========================================================
"Live Cell Fluorescent Imaging"

Presented by Nicholas Beavers, Media Cybernetics

Wednesday, 09-January at 1:30 PM (New York time)

==========================================================
"Tracking Objects in 2D and 3D"

Presented by Paul Jantzen, Media Cybernetics

Friday, 11-January at 1:30 PM (New York time)

==========================================================

Calendar, descriptions, and pre-registration is at:
http://www.magbiosystems.com/education

Seminars require that attendees use a Java-enabled browser with a high
bandwidth connection. Audio is via toll-free telephone.
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Webinar Invitation: Quantitative Imaging of Fluorescent Specimens

2008-01-02T16:28:55Z

You are invited to attend a live, interactive, web-based instructional
seminar:

==========================================================
"Quantitative Image and Data Acquisition for Fluorescent Specimens"
Advice from a Facility Director

Presented by Brian Matsumoto, Ph.D., University of California, Santa
Barbara
==========================================================

Details are below. Connection lines are limited so reserve yours now.
There is no charge to participate in this on-line seminar.

When:
=====================
Monday, 7-January, 1:30PM (New York time; 10:30 AM California time.)
Duration: Approximately 1 hour.

Pre-register (required) at:
http://magbiosystems.com/education/

Details:
=====================

Attendees will learn about issues affecting the quantitative accuracy
of fluorescence images acquired through a microscope and will pick up
tips and suggestions for improving image quality, making better use of
the camera's light collection abilities, and will learn the
nomenclature associated with digital imaging. Attendees will leave
with a better understanding of how to characterize and optimize their
optical system, camera, and software. Bring your questions to this
live, interactive web-based seminar.

- What is bit depth and when/why does it matter?
- What is dynamic range?
- Characterizing your camera's linear range.
- Noise and other sources of data uncertainty.
- Maximizing your camera's light collection capabilities.
- Setting the best exposures.
- Correcting for photobleaching.
- Collecting images in 3D.

About the presenter
=====================
Brian Matsumoto, Ph.D. is the Director of the Integrated Microscopy
Facility and is an Associate Adjunct Professor for the department of
Molecular, Cellular and Developmental Biology Department at the
University of California Santa Barbara. He is the author of "Basic
Methods in Light Microscopy" (Cambridge University
Press) and editor of "Cell Biological Applications of Confocal
Microscopy" (Academic Press).

More instructional webinars are also shown at http://magbiosystems.com/education
Register for any session of interest and feel encouraged to pass this
along to your colleagues.

Sponsored by the Microimaging Applications Group (MAG), a group of
independent imaging companies working cooperatively to provide an
unparalleled range of solutions for microimaging applications.

This seminar requires that attendees use a Java-enabled browser with a
high bandwidth connection. Audio is via toll-free telephone.

There is no charge to participate in this on-line seminar.
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Re: Dynamic quenching

2007-10-24T13:05:18Z

Hello everybody I need help. I was surfing the net for
this problem and came across this Bionet.

I have a question about fluorescence
dynamic/collisional quenching. I know in static
quenching fluorophore makes complex with quencher in
the ground state, the uncomplexed fluorophore
concentration decreases and hence emission intensity
decreases. In collisional quenching, fluorophore is
excited to the excited state and quencher diffuse to
the fluorophore during the lifetime of the fluorophore
and then fluorophore returns to the ground state
without emission, and hence emission intensity
decreases. From stern-volmer equation we know that
lifetime also decreases with quencher concentration.
But my question is Why/How the lifetime of the
fluorophore decreases and why the decrease in lifetime
is quencher dependent; quencher concentration inceases
lifetime of fluorophore decreases.

I really appretiate any comment/explanation.

Thank you very much
rahman

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CHI's In Vitro Molecular Imaging Conference, part of Imaging Week

2007-09-07T06:19:44Z

Conference Announcement:
CHI's Fourth Annual In Vitro Molecular Imaging
November 27-28, 2007
Hyatt Regency Mission Bay Spa & Marina - San Diego, CA
www.Imaging-Week.com

CHI's In Vitro Molecular Imaging, part of Imaging Week, will feature comprehensive discussions from industry and academic leaders on FRET, BRET, photoswitchable fluorescent, proteins, functional imaging, fluorescent probes for in vitro and in vivo, multicolor fluorescent proteins, cellular imaging, and imaging technologies. Stay on to attend our Fourth Annual In Vivo Molecular Imaging conference on November 29-30, 2007.

If you are interested in attending, be sure to register by September 14th to claim the early bird discount. Visit the event's website for full details and to register, or call CHI at 781-972-5400.

Sincerely,

James Prudhomme
Cambridge Healthtech Institute

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Webinar Invitation: Quantitative Image and Data Acquisition for Fluorescent Specimens

2007-09-05T17:32:21Z

You are invited to attend a live, interactive, web-based instructional
seminar:

==========================================================
"Quantitative Image and Data Acquisition for Fluorescent Specimens"
Advice from a Facility Director

Presented by Brian Matsumoto, Ph.D., University of California, Santa
Barbara
==========================================================

Details are below. Connection lines are limited, so reserve yours
now. There is no charge to participate in this on-line seminar.

When:
=====================
Thursday, 13-September, 1:00PM (Eastern time; 10 AM Pacific time.)
Duration: Approximately 1 hour.

Pre-register (required) at:
https://mediacy.webex.com/mediacy/j.php?ED=99592442&RG=1

[If this link has wrapped, please re-build it in your web browser's
address bar. The line begins with "https" and ends with "RG=1" ]

Details:
=====================

Attendees will learn about issues affecting the quantitative accuracy
of fluorescence images acquired through a microscope and will pick up
tips and suggestions for improving image quality, making better use of
the camera's light collection abilities, and will learn the
nomenclature associated with digital imaging. Attendees will leave
with a better understanding of how to characterize and optimize their
optical system, camera, and software. Bring your questions to this
live, interactive web-based seminar.

- What is bit depth and when/why does it matter?
- What is dynamic range?
- Characterizing your camera's linear range.
- Noise and other sources of data uncertainty.
- Maximizing your camera's light collection capabilities.
- Setting the best exposures.
- Correcting for photobleaching.
- Collecting images in 3D.

About the presenter
=====================
Brian Matsumoto, Ph.D. is the Director of the Integrated Microscopy
Facility and is an Associate Adjunct Professor for the department of
Molecular, Cellular and Developmental Biology Department at the
University of California Santa Barbara. He is the author of Basic
Methods in Light Microscopy (Cambridge University
Press) and editor of Cell Biological Applications of Confocal
Microscopy (Academic Press).

Upcoming Webinar: "Deconvolving 3D Fluorescence Images for
Quantitative Analysis" (invitations will be sent separately).

Sponsored by Media Cybernetics (Image Pro and AutoQuant software) and
QImaging (precision CCD cameras)

This seminar requires that attendees use a Java-enabled browser with a
high bandwidth connection. Audio is via toll-free telephone.

There is no charge to participate in this on-line seminar.

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Re: help for oocyte staining method

2007-07-15T04:18:54Z

Dear Berenice,

were you able to find anything in PubMed, eg by serching like this:
http://www.ncbi.nlm.nih.gov/sites/entrez?db=pubmed&cmd=search&term=oocyte%20mitochondria%20staining

or by googling

http://www.google.de/search?q=oocyte+mitochondria+staining&ie=utf-8&oe=utf-8&aq=t&rls=org.mozilla:en-US:official&client=firefox-a

Then there are mitochondrial stains, you might check http://www.probes.com

and proteins as prohibitin or SGK1 (antibodies available eg by www.abnova.com
and http://www.cellsignal.com)

you might cross.check bionet.molbio.methds-reagnts (http://
groups.google.com/group/bionet.molbio.methds-reagnts), we had a small
discussion on mitochondrial markers there recently. (As fluorpro has
not much activity, you might consider posting there, even your
question is not so straightforward molbio)

Best regards,

Wolfgang

On Jul 14, 2:47 am, "Berenice" <beren...@...> wrote:

> I work in Brazil with canine IVM and IV,and notice your mail at Fluorpro. I would apreciate very much if you could send me any information about oocyte mithocondria staining method, which might be useful to clarify some aspects in my resarch about this subject.
>
> I thank you very much for your attention to this request.
>
> Dr. Berenice de Avila Rodrigues
>
> Laboratório de Embriologia e Biotécnicas de Reprodução - UFRGS
>
> Caixa Postal 15004
>
> 91501-970 Porto Alegre, RS, Brazil
>
> e-mail: beren...@...

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Re: help for oocyte staining method

2007-07-13T18:47:43Z

I work in Brazil with canine IVM and IV,and notice your mail at Fluorpro. I would apreciate very much if you could send me any information about oocyte mithocondria staining method, which might be useful to clarify some aspects in my resarch about this subject.

I thank you very much for your attention to this request.

Dr. Berenice de Avila Rodrigues

Laboratório de Embriologia e Biotécnicas de Reprodução - UFRGS

Caixa Postal 15004

91501-970 Porto Alegre, RS, Brazil

e-mail: berenice@...

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Call for Speakers: Fluorescent Proteins Conference, Nov. 27-28

2007-05-25T08:42:45Z

Cambridge Healthtech Institute is accepting speaker proposals for the
Fourth Annual Fluorescent Proteins in Drug Discovery conference (part of
"Imaging Week"), scheduled for November 27-28, 2007, San Diego, CA.

If you would like to submit a proposal to give a presentation at this
meeting, please submit a title and brief (3 5 sentences) summary on your
recent work in the area of in fluorescent proteins on-line at
www.Imaging-Week.com by June 8, 2007.

Otherwise, email your speaker proposal to Margit Eder, Ph.D., Conference
Director: meder@...
If you would like to speak with Dr. Eder, she can be reached at 781-972-5478.

Regards,

James Prudhomme
Cambridge Healthtech Institute

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Re: stability of luciferase reagents?

2007-03-08T01:48:39Z

You could read the experimental procedures in one of my old papers : "
Stimolazione della formazione di AMP ciclico nel cervello di ratto "in
vitro" da parte della nicergolina" - Il Farmaco 31, 10-17 (1976)

Regards
PCM

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stability of luciferase reagents?

2007-03-07T06:48:23Z

Hi there,
I am a PhD student in Dortmund/Germany and would like to set up a
luciferase assay- does anyone have experience how to solve the
reagents (luciferase, D-luciferin sodium salt), how stable they are in
solution and if you can freeze stock solutions? And how stable the
bioluminescent signal is?
Thanks a lot in advance!

Uyen

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Thermostability of GFP

2007-02-12T03:23:14Z

Hello,

I’m a phD student from the University of Coimbra, Portugal.

Could you give me information about thermostability of GFP in Thermus

host? There are any reports of thermus transformation with plasmid/GFP.

Thanks a lot.

Susana Alarico

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About GFP-vectors!

2007-01-23T16:28:27Z

Hello, My name is Keith Smith. I am a senior at William Jewell College in Liberty, Missouri. I am looking for a GFP-vector that I can insert a gene of interest into and transfect into E. coli cells. I was wondering if anyone would be willing to donate some to my Molecular Genetics Lab at school. I am about to start a neat project, but I do not have the funds to buy one and my school is not going to provide any. Or does anyone know where or who I might be able to come across some GFPs? Thanks

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Anyone used Molecular Probes ALEXA dyes?

2006-12-05T10:20:32Z

Dear Janet,

I read a very old post from you answering questions about preventing Alexa 488 from fading. In it you said your samples last longer if you seal them. How do you seal them? Do you use nail polish?

Thanks very much,

Cynthia

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GFP-half life

2006-11-08T06:53:10Z

I saw the message posted on may 1996 and it's exactly the answer to these
questions I am looking for
I am trying to express a GFP-reporter construct in HeLa cells, and I am
wondering (1) what is the halflife of GFP in a mammalian cell, and (2) how long
does it take to get expression of GFP from the time of induction of the
construct?

Thanks in advance!

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GFP tagged receptor, signal quenching ?

2006-09-18T03:12:20Z

Dear All,

We have constructed a receptor bearing GFP and did the necessary test to
see if things works
(CHO transient expression works like a charm, nice signal on the FACS).
So next thing was to have 3T3 cells express the GFP tagged receptor
stably (G418 selection),
unfortunately the FACS signal is not as high as in the CHO expression
and in some cases downright poor.
Might it be possible that the receptor quenches the GFP signal or are we
facing other problems?

Thanks for your time.

--

Ing. Walter Pouwels
Academic Medical Center
University of Amsterdam
Laboratory for Experimental Immunology
G1-106
P.O. box 22700
1100 DE Amsterdam
The Netherlands

Tel.: +31-20-566768756/ 68036/ 5662285
Fax: +31-20-5669756

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pDsRed-express-C1

2006-06-16T13:45:09Z

Hi:

Does anyone has the Clontech vector pDsRed-express-C1, or has experience using it?

------------------------------------------------------------------------------------

Zhihua Han, PhD

Department of Biochemistry and Molecular Biology

East Tennessee State University

Johnson City, TN 37614

Phone:423-439-2128

Fax: 423-439-2030

------------------------------------------------------------------------------------

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nuclear GFP

2006-06-14T14:59:10Z

Hello,

I am looking for a nuclear GFP construct. We want to generate several cell lines that stably express nuclear GFP. We specifically require very uniform labeling of the nucleus. I was looking at the sales material for BD/Clontech's Living Colors contructs, which includes a photo of a pDsRed2-Nuc labeled cell. The web address for this sales note (and the image) is below:

http://wolfson.huji.ac.il/purification/PDF/Tag_Protein_Purification/FluorescentProteins/BD_AcGFP1MonFluorProt.pdf

This cell shows a very punctate nuclear labeling pattern. Is the patchy labeling evident in the Clontech image due to extra nucleolar labeling? I realize this is labelig with red instead of green, but I'm concerned the green construct could generate similar results. I've looked around at some images in the literature of nuclear GFP, and I didn't observe this kind of blotchy labeling. Is patchy labeling a common feature of nuclear targeted GFP? This would be a problem for us.

I have located a more "sophisticated" nuclear GFP sold through Qbiogene (pQBI-tatGFP, cat AFP 3202) that claims to label both the nucleus and nucleolus, thus, generating very uniform nuclear labeling. Would this construct be preferable if uniform labeling is a priority?

Additionally, I've read that prolonged GFP expression can be toxic to cells. I know that the many "new and improved" GFP variants today are supposedly less toxic, more bright, etc. etc. etc. than the original version. I have used several cell lines that stably expressed cytoplasmic GFP without any obvious effects on viability. Would long-term nuclear expression present more of a problem?

In summary, I would appreciate any feedback that anyone has on these two issues:

1. The uniformity of nuclear GFP expression

2. The toxicit of long-term nuclear GFP expression

Thank-you!!!

Beth

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GFP and in situ conditions

2006-06-06T19:11:08Z

Dear Peter,

Has anyone let you know a protocol for fluorescent in situ hybridization on GFP-labeled cells? I also would like to know it myself. Can you let me know? I also trying to locate Lac-z gene(from GT(rosa 26)mice) by FISH. Do you have any idea?

BTW I found soo many kits available in at probes.invitrogen.com

Nisha

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GFP

2006-05-09T08:53:34Z

Hi,
Im a final year sudent in university and am currently finishing my project
on synchronisation of plant cell suspension cultures. Im preforming a transformation
using Arabidopsis cells and and Agrobacterium strains ACD2 and GDR. My supervisor
said I was to test for transformation using GFP. I did this ana my cells
did fluoresce a green colour which I understand to be a positive for transformation.
What Im not clear about is whether the strains express GFP and this is why
they fluoresce. Can anyone help me with this?
Thanks
Anita

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Selecting a CCD Camera for Light Microscopy (Webinar invitation)

2006-04-17T15:43:56Z

"Selecting a CCD Camera for Light Microscopy," a live, informative, and
interactive web-based seminar is being held this Friday (21-April) at
11:30 AM (New York time).

Details are below. There is no cost, but connection lines are limited
so reserve yours now.

--------------------------------------------------------------
TO RESERVE YOUR CONNECTION LINE
--------------------------------------------------------------
Surf to this URL:

https://premconf.webex.com/premconf/j.php?ED=86778962&RG=1

Click REGISTER and complete the requested information. You will be sent
a link that gives you access to Friday's meeting.

----------------------------------------
MEETING SUMMARY
----------------------------------------
Name: Selecting a CCD Camera for Light Microscopy
Date: Friday, April 21, 2006
Time: 11:30 am, Eastern Daylight Time (New York).
Meeting Number: 742096476

Agenda:

-Understanding Digital Camera Parameters:
-Camera Sensitivity
-Noise Limitations
-Dynamic Range (Full-Well Capacity)
-Use of Gain
-Color Acquisition Options
-Note on Under- and Over-Sampling
-Matching Pixel Size to Optical Resolution
-Questions from the Audience

Presented by QImaging Corporation (http://www.qimaging.com), makers of
precision cameras for microscopy, the information imparted will be
broadly useful to anyone striving to make the best camera choice for
their imaging goals.

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pHygEGFP or pNeoEGFP

2006-04-10T08:50:19Z

Hi,
does anybody has a good or bad experience with any of these plasmids
expressing fusion proteins between EGFP and Neo or Hyg resistance.
Are both markers well expressed, in what cell lines, any toxicity...
I'm interested in using them in Jurkat cells after cloning the fusion
protein into a retroviral vector, under the control of the retroviral
promoter. Thanks in advance,
Albert

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Questions about DAPI staining with cryosection of mouse lung

2006-03-20T17:58:54Z

HI, Dear All,

This is Chengming from Auburn University, and this is my first time
here and I have a question to ask about the DAPI stain with cryosection
of mouse lung.

We created cryosection slides (no parafin fixing), and stained with
DAPI. The problem I had was that the shape of the nuclei was weird,
not round as seen the normal DAPI staining.

Sorry to bother you all, and your suggestions are highly appreciated.

Thanks,

Chengming

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Regulated proteins expression

2006-03-01T17:10:02Z

Dear Colleagues,

I'm looking for a way to regulate expression of two proteins
simultaneously. After transfection I need expression of only one of two
proteins of interest. After a while expression of the another should be
launched witth the first one blocked at the same time.

Will there be any suggestions on this specific regulation mechanism
development? There is no difference at which level to conduct the
regulation - transcription/translation, etc...

Thank you in advance,
Vladimir

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