Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi,
Could someone point me to a list of water-insoluble 405nm and 561
nm excitable fluorescent dyes (and typical suppliers: offline is ok)?
Thanks!
Anand
-----------------------------------------------------------------
Anand Yethiraj email: [log in to unmask]
Dept. of Physics and Phys. Oceanography phone: 709-737-2113
Memorial University of Newfoundland fax: 709-737-8739
St. John's, NL A1B 3X7, Canada. web: www.physics.mun.ca/~anand
-----------------------------------------------------------------
On Sat, 17 Mar 2007, CONFOCAL automatic digest system wrote:
> There are 7 messages totalling 1106 lines in this issue.
>
> Topics of the day:
>
> 1. precisExcite Illuminator
> 2. GFP in nuclei (3)
> 3. Question about saving Confocal Images On Leica SL
> 4. 96 well plates with filters (2)
>
> ----------------------------------------------------------------------
>
> Date: Fri, 16 Mar 2007 12:23:18 +0000
> From: James Kerin <[log in to unmask]>
> Subject: Re: precisExcite Illuminator
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> <html>
> <head>
> <!-- Written by GoldMine : Version 6.70.50123
> -->
> <title></title>
> </head>
> <BODY text="#000000" link="#0000FF">
> <font style="font-family:'Tahoma';font-size:10pt;">Regarding
> questions about recent LED technology:<p>
> The different LED colours are based on somewhat
> different semiconductor compositions, and
> right now it is the general case that LEDs
> designed to work in the green part of the
> spectrum are less efficient than those in
> the red and the blue. We may not notice this
> visually, as our eyes are much more sensitive
> in the green than in the red and (especially)
> blue parts of the spectrum, but it certainly
> does show up when you make quantitative measurements
> using a better detector (with apologies to
> any intelligent designers out there!) The
> maximum light output from an LED is ultimately
> limited by thermal considerations, and the
> steady improvements in LED intensity have
> reflected improvements in dissipating heat
> away from the LED chip (so that you can drive
> it harder) as well as those in their optical
> conversion efficiency.<br>
> <br>
> The thermal limitation means that if you
> don't need to have an LED on all the time,
> you can drive it with a correspondingly higher
> current before you reach the thermal limit,
> as it will take a while for the LED to heat
> up. The thermal time constant generally tends
> to be long enough to allow a useful degree
> of overdriving in situations where you are
> switching between LEDs to achieve alternations
> between different excitation wavelengths
> - which is exactly what you want to do in
> many cases anyway. A detailed analysis is
> potentially complex, but as a rule of thumb
> it seems generally safe to overdrive transiently
> in order to maintain the rated average current,
> for frequencies down to maybe around 10Hz
> or so, and in practice this does seem to
> be a conservative limit.<br>
> <br>
> This means that overdriving is feasible in
> many fluorescence applications. However,
> there is then another problem to be solved,
> but luckily that's quite easy. The problem
> is that if the LED is pulsed on and off,
> the chip temperature will be going up and
> down to some extent, and LEDs generally tend
> to become less efficient when they are hotter.
> Therefore the optical output may decline
> during the on periods as the chip heats up.
> The reduction may only be a few percent,
> but it can be eliminated for all practical
> purposes by using a photodiode in the LED
> assembly to provide an optical feedback signal.
> The LED is then driven so as to give constant
> optical output during the pulse, which can
> be done very accurately as the feedback can
> be made to operate in the microsecond range
> (in our OptoLED at least!).<br>
> <br>
> Optical feedback has another potential advantage
> too. It can be used to stabilise the LED
> output over arbitrarily long periods, as
> photodiodes are themselves inherently very
> stable. With a good enough feedback photodiode,
> you should be fighting the accuracy of your
> measurement system if you do try to measure
> the stability, so a figure of even as low
> as 0.1% may well be conservative. Way too
> low to worry about for fluorescence applications
> anyway!<p>
> <p>
> </font><font style="font-family:'Arial';font-size:10pt;"
> color="#0000FF">>Date: Wed, 14 Mar 2007
> 10:33:07 +0000<br>
> >From: Nuno Moreno <br>
> >Subject: precisExcite Illuminator<br>
> <br>
> >Hi!<br>
> <br>
> >As anyone ever tried precisExcite for
> fluorescence excitation. I'm very <br>
> >tempted to buy one but I still have some
> doubts about the practical <br>
> >usage of its intensity, specially in
> the green region.<br>
> <br>
> >Any comment is welcome,<br>
> >Nuno Moreno<br>
> <br>
> >------------------------------<br>
> <br>
> >Date: Wed, 14 Mar 2007 11:49:43 +0000<br>
> >From: John Runions <br>
> >Subject: Re: precisExcite Illuminator<br>
> <br>
> <br>
> >Hi Nuno,<br>
> >We have tested this system on both compound
> and stereo scopes.&nbsp; It<br>
> >worked very well for short wavelength/
> blue excitation on the compound<br>
> >microscope but I couldn't see GFP emission
> using a stereoscope with a<br>
> >1x objective.&nbsp; The significant
> problem with these systems is the lack<br>
> >of green excitation for red fluorescence
> so we are holding off for the<br>
> >time being.&nbsp; I would guess that
> they are working madly to perfect the<br>
> >green LED.<br>
> ><br>
> >Regards, John.<br>
> ><br>
> >Nuno Moreno wrote:<br>
> <br>
> <br>
> </font><font style="font-family:'Arial';font-size:10pt;"><br>
> <br>
> </font><font style="font-family:'Tahoma';font-size:10pt;">With
> thanks<p>
> Dr. Martin V Thomas<br>
> Managing Director<br>
> Cairn Research Ltd<br>
> Faversham<br>
> Kent<br>
> UK<br>
> </font><a href="mailto:[log in to unmask]"><font
> style="font-family:'Tahoma';font-size:10pt;">[log in to unmask]</font></a><font
> style="font-family:'Arial';font-size:10pt;"><br>
> </font><a href="http://www.cairn-research.co.uk"><font
> style="font-family:'Tahoma';font-size:10pt;">www.cairn-research.co.uk</font></a><font
> style="font-family:'Arial';font-size:10pt;"><br>
> <br>
> <br>
> <br>
> <br>
> <br>
> <p>
> <p>
> </font></body>
> </html>
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 17:08:48 +0100
> From: F Javier Diez Guerra <[log in to unmask]>
> Subject: GFP in nuclei
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>
> Hi,
>
> Anybody knows why EGFP (or any of its color=20
> variants) concentrates at cell nuclei when expressed in eukaryotic cells?
>
> I mean just EGFP, without any tag or fusion.
>
> My experience with XFPs derived from A. Victoria=20
> is that they show accumulation at cell nuclei.
>
> Thanks
>
>
> F Javier Diez-Guerra, PhD
> Profesor Titular
> Centro de Biologia Molecular Severo Ochoa
> Facultad de Ciencias, Universidad Aut=F3noma
> Ctra Colmenar Viejo Km 15
> Cantoblanco, 28049 Madrid
> SPAIN
>
> phone: +34 91 4978051
> Fax: +34 91 4978087
> e-mail: [log in to unmask]
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 11:30:13 -0500
> From: "Kurten, Richard C" <[log in to unmask]>
> Subject: Re: GFP in nuclei
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>
> My experience in cultured CV1 cells using confocal imaging is that the
> intensity of untagged XFP in the nucleus is the same as that in the
> cytoplasm. In wide-field it appears to accumulate because the nucleus is
> thicker than the cytoplasm as the cell spreads. Tagging the molecule so it =
> is
> bigger than 40-45kDa results in exclusion from the nucleus in both imaging
> modes.
>
> Richard C. Kurten Ph.D.
> Associate Professor, Physiology & Biophysics
> UAMS College of Medicine
> Co-Director, Lung Cell Biology Laboratory
> Arkansas Children's Hospital Research Institute
> Director, Digital and Confocal Microscopy Laboratory
> Arkansas Cancer Research Center
> Little Rock, Arkansas
> UAMS: 501-686-8269 ACHRI: 501-364-2823
>
>
> -----Original Message-----
> =46rom: Confocal Microscopy List [mailto:[log in to unmask]] On
> Behalf Of F Javier Diez Guerra
> Sent: Friday, March 16, 2007 11:09 AM
> To: [log in to unmask]
> Subject: GFP in nuclei
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa=3FS1=3Dconfocal
>
> Hi,
>
> Anybody knows why EGFP (or any of its color
> variants) concentrates at cell nuclei when expressed in eukaryotic cells=3F
>
> I mean just EGFP, without any tag or fusion.
>
> My experience with XFPs derived from A. Victoria is that they show
> accumulation at cell nuclei.
>
> Thanks
>
>
> =46 Javier Diez-Guerra, PhD
> Profesor Titular
> Centro de Biologia Molecular Severo Ochoa Facultad de Ciencias, Universidad
> Aut=F3noma Ctra Colmenar Viejo Km 15 Cantoblanco, 28049 Madrid SPAIN
>
> phone: +34 91 4978051
> =46ax: +34 91 4978087
> e-mail: [log in to unmask]
>
> Confidentiality Notice: This e-mail message, including any attachments, is =
> =66or the sole use of the intended recipient(s) and may contain confidentia=
> l=
> and privileged information. Any unauthorized review, use, disclosure or =
> distribution is prohibited. If you are not the intended recipient, please =
> contact the sender by reply e-mail and destroy all copies of the original =
> message.
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 13:11:53 -0400
> From: Carol Bayles <[log in to unmask]>
> Subject: Re: Question about saving Confocal Images On Leica SL
>
> --============_-1038057378==_ma============
> Content-Type: text/plain; charset="us-ascii" ; format="flowed"
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Michelle,
> I am guessing that your problem is that you saved the projection
> (flattened image) as a "snapshot" in Leica. go back and redo it and
> save it as 'raw' which is like a .tif and you will be able to open
> the image as a 12 bit image in Metamorph and do thresholding, etc.
>
> carol
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>> Hello Everyone,
>>
>> I have what I hope will be a quick question to answer. I have
>> acquired a stack of 12 bit images on the Leica SL. I have flattened
>> that stack into an averaged intensity image, but when I captured the
>> flattened "snapshot" it looses resolution, and I cannot threshold
>> the image in MetaMorph. Does anyone know how or if it is possible,
>> to save a flattened stack and maintain its bit depth?
>>
>> Thank you,
>>
>> Michelle Sutorik
>> UCSF Diabetes Center
>> Microscopy and Imaging Core Facility Manager
>> 513 Parnassus Ave
>> Medical Sciences S1230
>> San Francisco, CA 94143-1222
>>
>>
>>
>> The fish are biting.
>> <http://us.rd.yahoo.com/evt=49679/*http://searchmarketing.yahoo.com/arp/sponsoredsearch_v2.php?o=US2140&cmp=Yahoo&ctv=Q107Tagline&s=Y&s2=EM&b=50>Get
>> more visitors on your site using <
>> http://us.rd.yahoo.com/evt=49679/*http://searchmarketing.yahoo.com/arp/sponsoredsearch_v2.php?o=US2140&cmp=Yahoo&ctv=Q107Tagline&s=Y&s2=EM&b=50>Yahoo!
>> Search Marketing.
>
>
> --
> <><><><><><><><><><><><><><><><><><><><><><>
> Carol Bayles, Manager
> Microscopy, Imaging & Fluorimetry (MIF)
> Biotechnology Resource Center
> 160a Biotech Bldg
> 607-254-4860
> www.brc.cornell.edu
>
> Confocal and Multiphoton Microscopy
> Nanobiotechnology Center
> www.nbtc.cornell.edu
>
> Cornell University
> Ithaca NY 14853
>
>
>
> --============_-1038057378==_ma============
> Content-Type: text/html; charset="us-ascii"
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
> <!doctype html public "-//W3C//DTD W3 HTML//EN">
> <html><head><style type="text/css"><!--
> blockquote, dl, ul, ol, li { padding-top: 0 ; padding-bottom: 0 }
> --></style><title>Re: Question about saving Confocal Images On Leica
> SL</title></head><body>
> <div>Michelle,</div>
> <div>I am guessing that your problem is that you saved the projection
> (flattened image) as a "snapshot" in Leica. go
> back and redo it and save it as 'raw' which is like a .tif and you
> will be able to open the image as a 12 bit image in Metamorph and do
> thresholding, etc.</div>
> <div><br></div>
> <div>carol</div>
> <div><br></div>
> <blockquote type="cite" cite>Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal</blockquote>
> <blockquote type="cite" cite>Hello Everyone,</blockquote>
> <blockquote type="cite" cite> </blockquote>
> <blockquote type="cite" cite>I have what I hope will be a quick
> question to answer. I have acquired a stack of 12 bit images on
> the Leica SL. I have flattened that stack into an averaged
> intensity image, but when I captured the flattened "snapshot"
> it looses resolution, and I cannot threshold the image in MetaMorph.
> Does anyone know how or if it is possible, to save a flattened stack
> and maintain its bit depth?</blockquote>
> <blockquote type="cite" cite> </blockquote>
> <blockquote type="cite" cite>Thank you,</blockquote>
> <blockquote type="cite" cite> </blockquote>
> <blockquote type="cite" cite>Michelle Sutorik</blockquote>
> <blockquote type="cite" cite>UCSF Diabetes Center</blockquote>
> <blockquote type="cite" cite>Microscopy and Imaging Core Facility
> Manager</blockquote>
> <blockquote type="cite" cite>513 Parnassus Ave</blockquote>
> <blockquote type="cite" cite>Medical Sciences S1230</blockquote>
> <blockquote type="cite" cite>San Francisco, CA 94143-1222<br>
> </blockquote>
> <blockquote type="cite" cite> </blockquote>
> <blockquote type="cite" cite>
> <hr size="1"></blockquote>
> <blockquote type="cite" cite>The fish are biting.<br>
> <a
> href=
> "http://us.rd.yahoo.com/evt=49679/*http://searchmarketing.yahoo.com/arp/sponsoredsearch_v2.php?o=US2140&cmp=Yahoo&ctv=Q107Tagline&s=Y&s2=EM&b=50"><span
>> </span>Get more visitors</a> on your site using <a
> href=
> "
> http://us.rd.yahoo.com/evt=49679/*http://searchmarketing.yahoo.com/arp/sponsoredsearch_v2.php?o=US2140&cmp=Yahoo&ctv=Q107Tagline&s=Y&s2=EM&b=50"><span
>> </span>Yahoo! Search Marketing.</a></blockquote>
> <div><br></div>
> <div><br></div>
> <x-sigsep><pre>--
> </pre></x-sigsep>
> <div
>> <><><><><><><><><<span
>> </span
>> ><><><><><><><><><span
>> </span><><><><><></div>
> <div>Carol Bayles<i>,</i> Manager<i><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab></i></div>
> <div><i>Microscopy, Imaging & Fluorimetry
> (MIF)<x-tab>
> </x-tab></i></div>
> <div>Biotechnology Resource Center<x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab></div>
> <div>160a Biotech Bldg</div>
> <div>607-254-4860<x-tab>
> </x-tab><i><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab></i></div>
> <div>www.brc.cornell.edu</div>
> <div><i><x-tab>
> </x-tab><x-tab>
> </x-tab></i></div>
> <div><i>Confocal and Multiphoton Microscopy</i></div>
> <div>Nanobiotechnology Center</div>
> <div>www.nbtc.cornell.edu</div>
> <div><br></div>
> <div>Cornell University</div>
> <div>Ithaca NY 14853<x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab><x-tab>
> </x-tab></div>
> <div><br></div>
> <div><br></div>
> <div><br></div>
> </body>
> </html>
> --============_-1038057378==_ma============--
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 18:11:53 +0100
> From: F Javier Diez Guerra <[log in to unmask]>
> Subject: Re: GFP in nuclei
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>
> Richard,
>
> We use NIH-3T3 cells and see more concentration=20
> (intensity) of XFPs within nuclei using confocal imaging.
>
> Further, we do not see differences between=20
> monomeric (A206K) and non-monomeric versions. All them concentrate at=
> nuclei.
>
>
> At 17:30 16/03/2007, you wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>>
>> My experience in cultured CV1 cells using confocal imaging is that the
>> intensity of untagged XFP in the nucleus is the same as that in the
>> cytoplasm. In wide-field it appears to accumulate because the nucleus is
>> thicker than the cytoplasm as the cell spreads. Tagging the molecule so it=
> is
>> bigger than 40-45kDa results in exclusion from the nucleus in both imaging
>> modes.
>>
>> Richard C. Kurten Ph.D.
>> Associate Professor, Physiology & Biophysics
>> UAMS College of Medicine
>> Co-Director, Lung Cell Biology Laboratory
>> Arkansas Children's Hospital Research Institute
>> Director, Digital and Confocal Microscopy Laboratory
>> Arkansas Cancer Research Center
>> Little Rock, Arkansas
>> UAMS: 501-686-8269 ACHRI: 501-364-2823
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List [mailto:[log in to unmask]] On
>> Behalf Of F Javier Diez Guerra
>> Sent: Friday, March 16, 2007 11:09 AM
>> To: [log in to unmask]
>> Subject: GFP in nuclei
>>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>>
>> Hi,
>>
>> Anybody knows why EGFP (or any of its color
>> variants) concentrates at cell nuclei when expressed in eukaryotic cells?
>>
>> I mean just EGFP, without any tag or fusion.
>>
>> My experience with XFPs derived from A. Victoria is that they show
>> accumulation at cell nuclei.
>>
>> Thanks
>>
>>
>> F Javier Diez-Guerra, PhD
>> Profesor Titular
>> Centro de Biologia Molecular Severo Ochoa Facultad de Ciencias, Universidad
>> Aut=F3noma Ctra Colmenar Viejo Km 15 Cantoblanco, 28049 Madrid SPAIN
>>
>> phone: +34 91 4978051
>> Fax: +34 91 4978087
>> e-mail: [log in to unmask]
>>
>> Confidentiality Notice: This e-mail message,=20
>> including any attachments, is for the sole use=20
>> of the intended recipient(s) and may contain=20
>> confidential and privileged information. Any=20
>> unauthorized review, use, disclosure or=20
>> distribution is prohibited. If you are not the=20
>> intended recipient, please contact the sender by=20
>> reply e-mail and destroy all copies of the original message.
>
> F Javier Diez-Guerra, PhD
> Profesor Titular
> Centro de Biologia Molecular Severo Ochoa
> Facultad de Ciencias, Universidad Aut=F3noma
> Ctra Colmenar Viejo Km 15
> Cantoblanco, 28049 Madrid
> SPAIN
>
> phone: +34 91 4978051
> Fax: +34 91 4978087
> e-mail: [log in to unmask]
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 16:06:03 -0400
> From: Claire Brown <[log in to unmask]>
> Subject: 96 well plates with filters
>
> This is a multi-part message in MIME format.
>
> ------=_NextPart_000_0013_01C767E4.FF4D2A00
> Content-Type: text/plain;
> charset="us-ascii"
> Content-Transfer-Encoding: 7bit
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> I have a user who is using 96 well plates with filters to grow epithelial
> cells. They want to check transfection efficiency, but we are too far from
> the top and the bottom of the dish due to the way the filters sit about
> halfway up the wells in the plate. I think we are going to end up cutting
> out the filters and looking at the cells with a coverslip, but I was
> wondering if anyone on the list has come up with a better idea?
>
>
>
> Any suggestions are welcome.
>
>
>
> Claire
>
>
>
> _________________________________________________________________
>
> Claire M. Brown, PhD
>
> Life Sciences Complex Imaging Facility Director
>
> McGill University Department of Biochemistry
>
> <http://www.lifesciencescomplex.mcgill.ca/imaging>
> http://www.lifesciencescomplex.mcgill.ca/imaging
> <http://www.lifesciencescomplex.mcgill.ca>
>
>
>
>
> ------=_NextPart_000_0013_01C767E4.FF4D2A00
> Content-Type: text/html;
> charset="us-ascii"
> Content-Transfer-Encoding: quoted-printable
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
> <html xmlns:o=3D"urn:schemas-microsoft-com:office:office" =
> xmlns:w=3D"urn:schemas-microsoft-com:office:word" =
> xmlns=3D"http://www.w3.org/TR/REC-html40">
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>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'>I have a user who is using 96 well =
> plates
> with filters to grow epithelial cells. They want to check transfection
> efficiency, but we are too far from the top and the bottom of the dish =
> due to
> the way the filters sit about halfway up the wells in the plate. I think =
> we are
> going to end up cutting out the filters and looking at the cells with a
> coverslip, but I was wondering if anyone on the list has come up with a =
> better
> idea?<o:p></o:p></span></font></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'><o:p> </o:p></span></font></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'>Any suggestions are =
> welcome.<o:p></o:p></span></font></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'><o:p> </o:p></span></font></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'>Claire<o:p></o:p></span></font></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3D"Comic Sans MS"><span =
> style=3D'font-size:
> 12.0pt;font-family:"Comic Sans MS"'><o:p> </o:p></span></font></p>
>
> <p class=3Dsection1 style=3D'margin:0in;margin-bottom:.0001pt'><i><font =
> size=3D2
> color=3Dblack face=3DArial><span =
> style=3D'font-size:10.0pt;font-family:Arial;
> color:black;font-style:italic'>__________________________________________=
> _______________________</span></font></i><o:p></o:p></p>
>
> <p class=3Dsection1 style=3D'margin:0in;margin-bottom:.0001pt'><i><font =
> size=3D2
> color=3Dblack face=3DArial><span =
> style=3D'font-size:10.0pt;font-family:Arial;
> color:black;font-style:italic'>Claire M. Brown, =
> PhD</span></font></i><o:p></o:p></p>
>
> <p class=3Dsection1 style=3D'margin:0in;margin-bottom:.0001pt'><i><font =
> size=3D2
> color=3Dblack face=3DArial><span =
> style=3D'font-size:10.0pt;font-family:Arial;
> color:black;font-style:italic'>Life Sciences Complex Imaging Facility =
> Director</span></font></i><o:p></o:p></p>
>
> <p class=3Dsection1 style=3D'margin:0in;margin-bottom:.0001pt'><i><font =
> size=3D2
> color=3Dblack face=3DArial><span =
> style=3D'font-size:10.0pt;font-family:Arial;
> color:black;font-style:italic'>McGill University Department of =
> Biochemistry</span></font></i><o:p></o:p></p>
>
> <p class=3Dsection1 style=3D'margin:0in;margin-bottom:.0001pt'><i><font =
> size=3D2
> color=3Dblack face=3DArial><span =
> style=3D'font-size:10.0pt;font-family:Arial;
> color:black;font-style:italic'><a
> href=3D"http://www.lifesciencescomplex.mcgill.ca/imaging"><strong><b><fon=
> t
> face=3DArial><span =
> style=3D'font-family:Arial'>http://www.lifesciencescomplex.mcgill.ca/imag=
> ing</span></font></b></strong></a><a
> href=3D"http://www.lifesciencescomplex.mcgill.ca"></a></span></font></i><=
> o:p></o:p></p>
>
> <p class=3DMsoNormal><font size=3D3 face=3DArial><span =
> style=3D'font-size:12.0pt'><o:p> </o:p></span></font></p>
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>
> ------=_NextPart_000_0013_01C767E4.FF4D2A00--
>
> ------------------------------
>
> Date: Fri, 16 Mar 2007 15:24:11 -0500
> From: "Kurten, Richard C" <[log in to unmask]>
> Subject: Re: 96 well plates with filters
>
> This is a multi-part message in MIME format.
>
> ------_=_NextPart_001_01C76809.0E97ABE1
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> charset=us-ascii
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>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=3Dconfocal
>
> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa=3FS1=3Dconfocal=20
>
> I have taken filters from 24 well plates and placed intact on large
> rectangular coverslips (~60mmx22mm) on an inverted scope and have easily =
> seen
> GFP using a x20 PlanNEOFluar objective. Potentialy messy, but nondestructive
> and with care the inserts can be returned to the plates for culture. My
> =66ilters were glued to the bottom of the inserts, so working distance =
> wasn't
> an issue - I could even reach them with a x20 PlanApo as well.
>
> =20
>
> Richard C. Kurten Ph.D.
>
> Associate Professor, Physiology & Biophysics
>
> UAMS College of Medicine
>
> Co-Director, Lung Cell Biology Laboratory
>
> Arkansas Children's Hospital Research Institute
>
> Director, Digital and Confocal Microscopy Laboratory
>
> Arkansas Cancer Research Center
>
> Little Rock, Arkansas
>
> UAMS: 501-686-8269 ACHRI: 501-364-2823
>
> =20
>
> -----Original Message-----
> =46rom: Confocal Microscopy List [mailto:[log in to unmask]] On
> Behalf Of Claire Brown
> Sent: Friday, March 16, 2007 3:06 PM
> To: [log in to unmask]
> Subject: 96 well plates with filters
>
> =20
>
> I have a user who is using 96 well plates with filters to grow epithelial
> cells. They want to check transfection efficiency, but we are too far from
> the top and the bottom of the dish due to the way the filters sit about
> halfway up the wells in the plate. I think we are going to end up cutting =
> out
> the filters and looking at the cells with a coverslip, but I was wondering =
> if
> anyone on the list has come up with a better idea=3F
>
> =20
>
> Any suggestions are welcome.
>
> =20
>
> Claire
>
> =20
>
> _________________________________________________________________
>
> Claire M. Brown, PhD
>
> Life Sciences Complex Imaging Facility Director
>
> McGill University Department of Biochemistry
>
> http://www.lifesciencescomplex.mcgill.ca/imaging
> <http://www.lifesciencescomplex.mcgill.ca/imaging>
> <http://www.lifesciencescomplex.mcgill.ca>=20
>
> =20
>
>
> Confidentiality Notice: This e-mail message, including any attachments, is =
> =66or the sole use of the intended recipient(s) and may contain confidentia=
> l=
> and privileged information. Any unauthorized review, use, disclosure or =
> distribution is prohibited. If you are not the intended recipient, please =
> contact the sender by reply e-mail and destroy all copies of the original =
> message.
>
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> Search the CONFOCAL archive at
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> Search the CONFOCAL archive at
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>
> <div class=3DSection1>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>I have taken filters from 24 well plates and place=
> d=
> intact
> on large rectangular coverslips (~60mmx22mm) on an inverted scope and have =
> easily
> seen GFP using a x20 PlanNEOFluar objective. Potentialy messy, but =
> nondestructive
> and with care the inserts can be returned to the plates for culture. My =
> =66ilters
> were glued to the bottom of the inserts, so working distance wasn’t an
> issue - I could even reach them with a x20 PlanApo as =
> well.</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'> </span></font></p>
>
> <div>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>Richard C. Kurten Ph.D.</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>Associate Professor, Physiology & =
> Biophysics</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'> UAMS College of Medicine</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>Co-Director, Lung Cell Biology =
> Laboratory</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'> Arkansas Children's Hospital Research =
> Institute</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>Director, Digital and Confocal Microscopy =
> Laboratory</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'> Arkansas Cancer Research =
> Center</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>Little Rock, Arkansas</span></font></p>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'>UAMS: </span></font><font
> size=3D2 color=3Dnavy><span =
> style=3D'font-size:10.0pt;color:navy'>501-686-8269</span></font><font
> size=3D2 color=3Dnavy><span style=3D'font-size:10.0pt;color:navy'> ACHRI: =
> </span></font><font size=3D2 color=3Dnavy><span =
> style=3D'font-size:10.0pt;color:navy'>501-364-2823</span></font></p>
>
> </div>
>
> <p class=3DMsoNormal><font size=3D2 color=3Dnavy face=3DArial><span =
> style=3D'font-size:
> 10.0pt;color:navy'> </span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D2 =
> =66ace=3DTahoma><span
> style=3D'font-size:10.0pt;font-family:Tahoma'>-----Original Message-----<br>
> <b><span style=3D'font-weight:bold'>From:</span></b> Confocal Microscopy =
> List
> [mailto:[log in to unmask]] <b><span =
> style=3D'font-weight:bold'>On
> Behalf Of </span></b>Claire Brown<br>
> <b><span style=3D'font-weight:bold'>Sent:</span></b> Friday, March 16, 2007=
> =
> 3:06
> PM<br>
> <b><span style=3D'font-weight:bold'>To:</span></b> =
> [log in to unmask]<br>
> <b><span style=3D'font-weight:bold'>Subject:</span></b> 96 well plates with
> =66ilters</span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 =
> =66ace=3DArial><span
> style=3D'font-size:12.0pt'> </span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans MS"'>I have a user who is=
> =
> using
> 96 well plates with filters to grow epithelial cells. They want to check
> transfection efficiency, but we are too far from the top and the bottom of =
> the
> dish due to the way the filters sit about halfway up the wells in the plate=
> .=
> I
> think we are going to end up cutting out the filters and looking at the =
> cells
> with a coverslip, but I was wondering if anyone on the list has come up wit=
> h=
> a
> better idea=3F</span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans =
> MS"'> </span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans MS"'>Any suggestions are
> welcome.</span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans =
> MS"'> </span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans =
> MS"'>Claire</span></font></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 face=3D"Comi=
> c=
> Sans MS"><span
> style=3D'font-size:12.0pt;font-family:"Comic Sans =
> MS"'> </span></font></p>
>
> <p class=3Dsection1 =
> style=3D'margin-right:0in;margin-bottom:0in;margin-left:.5in;
> margin-bottom:.0001pt'><i><font size=3D2 color=3Dblack face=3DArial><span
> style=3D'font-size:10.0pt;font-family:Arial;color:black;font-style:italic'>=
> _________________________________________________________________</span></f=
> ont></i></p>
>
> <p class=3Dsection1 =
> style=3D'margin-right:0in;margin-bottom:0in;margin-left:.5in;
> margin-bottom:.0001pt'><i><font size=3D2 color=3Dblack face=3DArial><span
> style=3D'font-size:10.0pt;font-family:Arial;color:black;font-style:italic'>=
> Claire
> M. Brown, PhD</span></font></i></p>
>
> <p class=3Dsection1 =
> style=3D'margin-right:0in;margin-bottom:0in;margin-left:.5in;
> margin-bottom:.0001pt'><i><font size=3D2 color=3Dblack face=3DArial><span
> style=3D'font-size:10.0pt;font-family:Arial;color:black;font-style:italic'>=
> Life
> Sciences Complex Imaging Facility Director</span></font></i></p>
>
> <p class=3Dsection1 =
> style=3D'margin-right:0in;margin-bottom:0in;margin-left:.5in;
> margin-bottom:.0001pt'><i><font size=3D2 color=3Dblack face=3DArial><span
> style=3D'font-size:10.0pt;font-family:Arial;color:black;font-style:italic'>=
> McGill
> University Department of Biochemistry</span></font></i></p>
>
> <p class=3Dsection1 =
> style=3D'margin-right:0in;margin-bottom:0in;margin-left:.5in;
> margin-bottom:.0001pt'><i><font size=3D2 color=3Dblack face=3DArial><span
> style=3D'font-size:10.0pt;font-family:Arial;color:black;font-style:italic'>=
> <a
> href=3D"http://www.lifesciencescomplex.mcgill.ca/imaging"><strong><b><font
> =66ace=3DArial><span =
> style=3D'font-family:Arial'>http://www.lifesciencescomplex.mcgill.ca/imagin=
> g</span></font></b></strong></a><a
> href=3D"http://www.lifesciencescomplex.mcgill.ca"></a></span></font></i></p>
>
> <p class=3DMsoNormal style=3D'margin-left:.5in'><font size=3D3 =
> =66ace=3DArial><span
> style=3D'font-size:12.0pt'> </span></font></p>
>
> </div>
>
>
> <P><pre wrap>Confidentiality Notice: This e-mail message, including any<cr>=
> =
> attachments, is for the sole use of the intended<cr> recipient(s) and may =
> contain confidential and privileged<cr> information. Any unauthorized =
> review, use, disclosure or<cr> distribution is prohibited. If you are not =
> the<cr> intended recipient, please contact the sender by reply<cr> e-mail =
> and destroy all copies of the original message.<cr>
> </pre></P></body>
>
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>
> ------_=_NextPart_001_01C76809.0E97ABE1--
>
> ------------------------------
>
> End of CONFOCAL Digest - 15 Mar 2007 to 16 Mar 2007 (#2007-65)
> **************************************************************
>
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