colocalization question
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simon walker (BI) |
Re: Input on the Leica STED
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They're obviously using the term in a different context now.
From the Leica website: High Speed and High Resolution - All in One The Leica Tandem Scanner combines two technological solutions in one system: a conventional scanning system - ideal for morphological imaging and three dimensional structure analyses, and a resonant scanning system - the best solution for high speed confocal imaging. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 12:04 To: [hidden email] Subject: Re: Input on the Leica STED Well, tandem scanning was the term coined by Mojimir Petran for his spinning-disk confocal system. Ever since then there have been companies offering similar systems. Leica used to offer such a system (based on the BD CARV) but have since discontinued it. But it is something very different in all respects (including both price and performance) from the Leica STED which is a super- resolution system offering 85nm resolution. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 10:46 PM To: [hidden email] Subject: Re: Input on the Leica STED Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..? -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 11:41 To: [hidden email] Subject: Re: Input on the Leica STED Hello, what are we talking about here? I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner. Please clarify! I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow. I do realise both are a bit remote from Brazil. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 9:37 PM To: [hidden email] Subject: Input on the Leica STED Hi, I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons). Many thanks ! Renato Renato A. Mortara Parasitology Division UNIFESP - Escola Paulista de Medicina Rua Botucatu, 862, 6th floor São Paulo, SP 04023-062 Brazil Phone: 55 11 5579-8306 Fax: 55 11 5571-1095 email: [hidden email] home page: www.ecb.epm.br/~ramortara No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM |
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In reply to this post by Renato A. Mortara
So, what do you want feedback about? I've used the STED system
and it's really great - I'm happy to give more details if that's what you are interested in. The Leica (CARV) tandem scanner I've never used (though I've played briefly with a BD CARV). I also haven't used the Leica resonant scanner though I'd quite like to. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 11:02 PM To: [hidden email] Subject: Clarifying ? Input on the Leica STED Hope it is slightly clearer... Leica is offering the STED system that that can be an option to their TCS SP5 confocal. Besides, they also offer a tandem scanner for the same instrument, presumably to achieve higher acquisitions speed. Thanks again ! Renato -----Mensagem original----- De: Confocal Microscopy List [mailto:[hidden email]] Em nome de simon walker (BI) Enviada em: sexta-feira, 6 de fevereiro de 2009 09:46 Para: [hidden email] Assunto: Re: Input on the Leica STED Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..? -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 11:41 To: [hidden email] Subject: Re: Input on the Leica STED Hello, what are we talking about here? I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner. Please clarify! I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow. I do realise both are a bit remote from Brazil. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 9:37 PM To: [hidden email] Subject: Input on the Leica STED Hi, I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons). Many thanks ! Renato Renato A. Mortara Parasitology Division UNIFESP - Escola Paulista de Medicina Rua Botucatu, 862, 6th floor São Paulo, SP 04023-062 Brazil Phone: 55 11 5579-8306 Fax: 55 11 5571-1095 email: [hidden email] home page: www.ecb.epm.br/~ramortara No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM |
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In reply to this post by simon walker (BI)
Well, maybe Moji will sue - I believe he registered a trademark!
Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 11:08 PM To: [hidden email] Subject: Re: Input on the Leica STED They're obviously using the term in a different context now. From the Leica website: High Speed and High Resolution - All in One The Leica Tandem Scanner combines two technological solutions in one system: a conventional scanning system - ideal for morphological imaging and three dimensional structure analyses, and a resonant scanning system - the best solution for high speed confocal imaging. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 12:04 To: [hidden email] Subject: Re: Input on the Leica STED Well, tandem scanning was the term coined by Mojimir Petran for his spinning-disk confocal system. Ever since then there have been companies offering similar systems. Leica used to offer such a system (based on the BD CARV) but have since discontinued it. But it is something very different in all respects (including both price and performance) from the Leica STED which is a super- resolution system offering 85nm resolution. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 10:46 PM To: [hidden email] Subject: Re: Input on the Leica STED Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..? -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 11:41 To: [hidden email] Subject: Re: Input on the Leica STED Hello, what are we talking about here? I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner. Please clarify! I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow. I do realise both are a bit remote from Brazil. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 9:37 PM To: [hidden email] Subject: Input on the Leica STED Hi, I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons). Many thanks ! Renato Renato A. Mortara Parasitology Division UNIFESP - Escola Paulista de Medicina Rua Botucatu, 862, 6th floor São Paulo, SP 04023-062 Brazil Phone: 55 11 5579-8306 Fax: 55 11 5571-1095 email: [hidden email] home page: www.ecb.epm.br/~ramortara No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM |
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Leica is using the term "tandem scanner" since 4 years now, so I don't
think they got sue. Michael > Well, maybe Moji will sue - I believe he registered a trademark! > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Electron Microscope Unit, Madsen Building F09, > University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of simon walker (BI) > Sent: Friday, 6 February 2009 11:08 PM > To: [hidden email] > Subject: Re: Input on the Leica STED > > They're obviously using the term in a different context now. > From the Leica website: > > High Speed and High Resolution - All in One The Leica Tandem Scanner > combines two technological solutions in one system: a conventional > scanning system - ideal for morphological imaging and three dimensional > structure analyses, and a resonant scanning system - the best solution for > high speed confocal imaging. > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Guy Cox > Sent: 06 February 2009 12:04 > To: [hidden email] > Subject: Re: Input on the Leica STED > > Well, tandem scanning was the term coined by Mojimir Petran for his > spinning-disk confocal system. Ever since then there have been companies > offering similar systems. Leica used to offer such a system (based on the > BD CARV) but have since discontinued it. But it is something very > different in all respects (including both price and performance) from the > Leica STED which is a super- resolution system offering 85nm resolution. > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, > Madsen Building F09, University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of simon walker (BI) > Sent: Friday, 6 February 2009 10:46 PM > To: [hidden email] > Subject: Re: Input on the Leica STED > > Presumably we're talking about the SP5 with conventional and resonant > scanners and adapted for STED..? > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Guy Cox > Sent: 06 February 2009 11:41 > To: [hidden email] > Subject: Re: Input on the Leica STED > > Hello, what are we talking about here? I've used the Leica STED (and I'm > seriously impressed with it) but I didn't know Leica made a tandem > scanner. Please clarify! I'm presenting results from the Leica STED at > NZCM next week in Rotorua and in April at FOM in Krakow. I do realise > both are a bit remote from Brazil. > > > Guy > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, > Madsen Building F09, University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Renato Mortara > Sent: Friday, 6 February 2009 9:37 PM > To: [hidden email] > Subject: Input on the Leica STED > > Hi, > > I would appreciate any input on the Leica Tandem Scanner confocal > microscope (pros and cons). > > Many thanks ! > > Renato > > Renato A. Mortara > Parasitology Division > UNIFESP - Escola Paulista de Medicina > Rua Botucatu, 862, 6th floor > São Paulo, SP > 04023-062 > Brazil > Phone: 55 11 5579-8306 > Fax: 55 11 5571-1095 > email: [hidden email] > home page: www.ecb.epm.br/~ramortara > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM |
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Daniel James White |
Re: colocalization question
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In reply to this post by Jean-Pierre CLAMME-2
Hi JP,
If you don't mind, I will also reply to the list, so the info i give is available to all.... There are 2 separate issues here: 1) Setting the PMT offset during imaging on a confocal laser scanning microscoipe 2) calculating thresholds for each channel in order to calculate "thresholded" Manders coefficients etc. I will deal with them separately.... 1) Only the person who made the sample knows what is real signal and what is background. Background can be any or all of the following: a) electronic noise from the detector b) non specific fluorescent antiobody staining (can be reduced by blocking non specific binding sites) c) non interesting signal in a non interesting part of the sample. etc. In order to use the full dynamic range of the analogue to digital converter ( that comes afrter the PMT detector, and turns the analogue signal from the PMT into the numbers that are displayed as colours on the computer screen) it is sensible to set the PMT offset voltage such that whatever tou decide is background in the sample has a signal value close to zero but just above zero. Using Hi Lo or range indicator LUT / palette you can do this easily, bt making background have some zero pixels and some pixels above zero. I generally aim for just less than 50% zero pixels. Some people are more conservative than that. The point is not to waste lots of your dynamic range by having the offset for too high, so your image values start at eg 30 instead of close to Zero. This can easily be seen in the image intensity histogram. The images should never be saturated, as this is throwing away usually the most interesting and important info. the brightest stuff is ususally what you are most interested in. so clipping that info is a really dumb thing to so. to see the lower iuntensities clearly, that are usually close to black and therefore very hard to see, simply use a rainbow or other colourful look up table / pallette, and you will see lower intensity details without having to saturate the image by increasing PMT gain and or laser power . Black to some colour look up tables are a really dumb way of visualising single channel image data. Hard scientists laugh at us for doing that, and they always use colourful look up tables / palettes. Just because a dye looks green ish through the eye piece doesnt mean it is really green (its some mixture of greenish wavelengths), so displaying it as (R G B) (0 255 0) is formally incorrect anyway. Better to represent intensities as different colours than grey scale or single colour brightness scale. If image data has a very high offset (data only starts a eg 30 or 50 etc , not close to 0 intensity) , then the auto threshold method of Costes fails, because it thinks image intensity of 30 is some real intensity it needs to take care of , when in fact this is just a uniform "background" intensity which is close to zero in reality. In this case it assumes the high background from the incorrectly set offset is real signal, and since it is present probably in both colour channels, of course it sees that there is intensity in both colour channels for every pixel in the background. Then it sets the thresholds below the lowest intensity values in the image and nearly ALL the image pixels are said to be colocalised, which is of course totally wrong! Better to set the PMT offset correctly during imaging, than manually subtract high background during image processing!!! Saturated images also confuse the software/algorithms: Very bright pixels having intensity value of 255 might really be any value over 255, and you cant know what the real value should have been. The real info was never collected. no amount of image processing can get it back! This then gives wrong results during intensity correlation calculation a la Pearson and Manders. Image saturation is BAD BAD BAD. 2) Simply use the method of Costes et al. This is simple, reproducible, not subjective and pretty robust. It will fail if background intensity is high (eg wrongly set PMT offset), and it will fail of there is really no spatial correlation in the intensity of the 2 dyes in question, because it is looking for a correlation that is not there. This is a sign of a clearly negative colocalisation result. This method is available in the imageJ plugin "Colocalization threshold" (not so user friendly) and in BioImageXD (hopefully more user friendly) You might get very slightly different results with these, as they are implemented in different programming languages and slightly differently. As a final note: Watch out - Manders coefficients are sensitive to signal:noise ratio. Noisier images - lower Manders coefficients. There exists a method to correct for that..... cheers Dan On Feb 6, 2009, at 1:34 AM, [hidden email] wrote: > > Hi, > > I have quick question! What do you suggest concerning tresholding > for colocalization studies ? I was always told, when I'm taking the > images from a confocal using the hi/low lut, to avoid saturation but > also to cut noise using the offset. However the specialist doing the > demo when installing our new confocal told me he never cut the noise > to remove noise. > > What do you think about this ? > > Best regards, > > JP > > > > - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - > > > Dan White <[hidden email]> wrote on 02/05/2009 03:08:16 AM: > > > Hi JP, > > > Here attached are the two papers you need to read, digest and > > understand. > > > BioImageXD is the easiest tool to use for this analysis (one reason > > why we made it) > > If you need help with that just ask. > > > In short, load the data, > > select the 2 channels in the file tree (using shift click on the > > second one) > > > click the colocalisation task icon (second icon in the midle set at > > the top) > > > click auto threshold > > click statistics, > > choose Costes method and calculate statistical significance. > > > You can export the results and the 2D scatter plot, > > THe 2D scatter plot / 2D histogram / fluorogram is far superior to a > > colour merge image (BAD BAD BAD!!!) to understand the spatial > > correlation of the intensity in the 2 data channels. > > It works very much lice a FACS / cytometer 2D hiostogram. > > Colour merge images are pretty, but misleading and generally evil, > in > > my opinion. > > > cheers > > > Dan > > > > > [attachment "Costes_etalColoc.pdf" deleted by jeanpierre clamme/ > NITTO] > > [attachment "manders.pdf" deleted by jeanpierre clamme/NITTO] > > > > Dr. Daniel James White BSc. (Hons.) PhD > > Senior Microscopist / Image Processing and Analysis > > Light Microscopy Facility > > Max Planck Institute of Molecular Cell Biology and Genetics > > Pfotenhauerstrasse 108 > > 01307 DRESDEN > > Germany > > > > > New Mobile Number!!! > > > +49 (0)15114966933 (German Mobile) > > +49 (0)351 210 2627 (Work phone at MPI-CBG) > > +49 (0)351 210 1078 (Fax MPI-CBG LMF) > > > http://www.bioimagexd.net > > http://www.chalkie.org.uk > > [hidden email] > > ( [hidden email] ) Dr. Daniel James White BSc. (Hons.) PhD Senior Microscopist / Image Processing and Analysis Light Microscopy Facility Max Planck Institute of Molecular Cell Biology and Genetics Pfotenhauerstrasse 108 01307 DRESDEN Germany New Mobile Number!!! +49 (0)15114966933 (German Mobile) +49 (0)351 210 2627 (Work phone at MPI-CBG) +49 (0)351 210 1078 (Fax MPI-CBG LMF) http://www.bioimagexd.net http://www.chalkie.org.uk [hidden email] ( [hidden email] ) |
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Megan Nicholson |
Re: Gelatin/Lysine/Collagen/Etc.
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In reply to this post by B. Prabhakar Pandian
Have you tried using plastic slides? Most adherent cells really grow well
on plastic. ibidi sells plastic slides with the optical quality of glass. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of B. Prabhakar Pandian Sent: Thursday, February 05, 2009 10:48 AM To: [hidden email] Subject: Gelatin/Lysine/Collagen/Etc. Hello, Does anybody have advice on what substrates (Gelatin/Lysine/Collagen/Etc.) is best for growing adherent cells on glass slides. Is there a study where cell differentiation was compared on various substrates. Any help would be great. Thanks, -Prabhakar |
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In reply to this post by Guy Cox
I have "played with" the Leica resonant scanner and I was quite impressed. The imaging is very fast and the images are relatively resolved given the speed. The system is also very easy to use and allows you to average and to utilize an 8kx8K resolution which cleans up the image considerably. However, once you perform considerable averaging and use such a high resolution you lose the speed. The take home message is that you have a lot of flexibility to perform either; very fast somewhat noisy imaging, or fast and somewhat resolved imaging, and everywhere in between. What settings you choose depends entirely on what you want to accomplish.
Cheers, Brian D Armstrong PhD Light Microscopy Core Manager Beckman Research Institute City of Hope Dept of Neuroscience 1450 E Duarte Rd Duarte, CA 91010 626-256-4673 x62872 http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: Friday, February 06, 2009 4:13 AM To: [hidden email] Subject: Re: Input on the Leica STED Well, maybe Moji will sue - I believe he registered a trademark! Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 11:08 PM To: [hidden email] Subject: Re: Input on the Leica STED They're obviously using the term in a different context now. From the Leica website: High Speed and High Resolution - All in One The Leica Tandem Scanner combines two technological solutions in one system: a conventional scanning system - ideal for morphological imaging and three dimensional structure analyses, and a resonant scanning system - the best solution for high speed confocal imaging. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 12:04 To: [hidden email] Subject: Re: Input on the Leica STED Well, tandem scanning was the term coined by Mojimir Petran for his spinning-disk confocal system. Ever since then there have been companies offering similar systems. Leica used to offer such a system (based on the BD CARV) but have since discontinued it. But it is something very different in all respects (including both price and performance) from the Leica STED which is a super- resolution system offering 85nm resolution. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 10:46 PM To: [hidden email] Subject: Re: Input on the Leica STED Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..? -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 11:41 To: [hidden email] Subject: Re: Input on the Leica STED Hello, what are we talking about here? I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner. Please clarify! I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow. I do realise both are a bit remote from Brazil. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 9:37 PM To: [hidden email] Subject: Input on the Leica STED Hi, I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons). Many thanks ! Renato Renato A. Mortara Parasitology Division UNIFESP - Escola Paulista de Medicina Rua Botucatu, 862, 6th floor São Paulo, SP 04023-062 Brazil Phone: 55 11 5579-8306 Fax: 55 11 5571-1095 email: [hidden email] home page: www.ecb.epm.br/~ramortara No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM --------------------------------------------------------------------- SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender. --------------------------------------------------------------------- |
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Dr. Mark A. DeCoster |
Re: Gelatin/Lysine/Collagen/Etc.
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In reply to this post by Megan Nicholson
Poly-lysine is a good one to try first and may be the easiest to work with.
As suggested cells grow better on plastic than on glass, but the optics are better on glass. -MD Dr. Mark A. DeCoster Associate Professor Biomedical Engineering Louisiana Tech University http://www2.latech.edu/~decoster/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Megan Nicholson Sent: Friday, February 06, 2009 8:38 AM To: [hidden email] Subject: Re: Gelatin/Lysine/Collagen/Etc. Have you tried using plastic slides? Most adherent cells really grow well on plastic. ibidi sells plastic slides with the optical quality of glass. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of B. Prabhakar Pandian Sent: Thursday, February 05, 2009 10:48 AM To: [hidden email] Subject: Gelatin/Lysine/Collagen/Etc. Hello, Does anybody have advice on what substrates (Gelatin/Lysine/Collagen/Etc.) is best for growing adherent cells on glass slides. Is there a study where cell differentiation was compared on various substrates. Any help would be great. Thanks, -Prabhakar |
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In reply to this post by Armstrong, Brian
I wasn't suggesting the Leica resonant scanner wasn't good!
Leaving aside the banter about Leica's 'illegitimate' use of the term 'tandem scanner', there is another point that's worth making here, and that is that it's a bad idea to get too many different features on the one stand. In Renato's example, a very expensive STED super-resolution system could be at risk of standing idle for a large part of the time while people use the resonant scanner for live-cell experiments. This can get very frustrating, and it's hardly sensible use of a million-dollar-plus STED instrument. Far better to scrape together (relatively) few additional dollars to set up a package deal on two instruments each optimised for one task. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Armstrong, Brian Sent: Saturday, 7 February 2009 4:07 AM To: [hidden email] Subject: Re: Input on the Leica STED I have "played with" the Leica resonant scanner and I was quite impressed. The imaging is very fast and the images are relatively resolved given the speed. The system is also very easy to use and allows you to average and to utilize an 8kx8K resolution which cleans up the image considerably. However, once you perform considerable averaging and use such a high resolution you lose the speed. The take home message is that you have a lot of flexibility to perform either; very fast somewhat noisy imaging, or fast and somewhat resolved imaging, and everywhere in between. What settings you choose depends entirely on what you want to accomplish. Cheers, Brian D Armstrong PhD Light Microscopy Core Manager Beckman Research Institute City of Hope Dept of Neuroscience 1450 E Duarte Rd Duarte, CA 91010 626-256-4673 x62872 http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: Friday, February 06, 2009 4:13 AM To: [hidden email] Subject: Re: Input on the Leica STED Well, maybe Moji will sue - I believe he registered a trademark! Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 11:08 PM To: [hidden email] Subject: Re: Input on the Leica STED They're obviously using the term in a different context now. From the Leica website: High Speed and High Resolution - All in One The Leica Tandem Scanner combines two technological solutions in one system: a conventional scanning system - ideal for morphological imaging and three dimensional structure analyses, and a resonant scanning system - the best solution for high speed confocal imaging. -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 12:04 To: [hidden email] Subject: Re: Input on the Leica STED Well, tandem scanning was the term coined by Mojimir Petran for his spinning-disk confocal system. Ever since then there have been companies offering similar systems. Leica used to offer such a system (based on the BD CARV) but have since discontinued it. But it is something very different in all respects (including both price and performance) from the Leica STED which is a super- resolution system offering 85nm resolution. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of simon walker (BI) Sent: Friday, 6 February 2009 10:46 PM To: [hidden email] Subject: Re: Input on the Leica STED Presumably we're talking about the SP5 with conventional and resonant scanners and adapted for STED..? -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Guy Cox Sent: 06 February 2009 11:41 To: [hidden email] Subject: Re: Input on the Leica STED Hello, what are we talking about here? I've used the Leica STED (and I'm seriously impressed with it) but I didn't know Leica made a tandem scanner. Please clarify! I'm presenting results from the Leica STED at NZCM next week in Rotorua and in April at FOM in Krakow. I do realise both are a bit remote from Brazil. Guy Optical Imaging Techniques in Cell Biology by Guy Cox CRC Press / Taylor & Francis http://www.guycox.com/optical.htm ______________________________________________ Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope Unit, Madsen Building F09, University of Sydney, NSW 2006 ______________________________________________ Phone +61 2 9351 3176 Fax +61 2 9351 7682 Mobile 0413 281 861 ______________________________________________ http://www.guycox.net -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Renato Mortara Sent: Friday, 6 February 2009 9:37 PM To: [hidden email] Subject: Input on the Leica STED Hi, I would appreciate any input on the Leica Tandem Scanner confocal microscope (pros and cons). Many thanks ! Renato Renato A. Mortara Parasitology Division UNIFESP - Escola Paulista de Medicina Rua Botucatu, 862, 6th floor São Paulo, SP 04023-062 Brazil Phone: 55 11 5579-8306 Fax: 55 11 5571-1095 email: [hidden email] home page: www.ecb.epm.br/~ramortara No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM --------------------------------------------------------------------- SECURITY/CONFIDENTIALITY WARNING: This message and any attachments are intended solely for the individual or entity to which they are addressed. This communication may contain information that is privileged, confidential, or exempt from disclosure under applicable law (e.g., personal health information, research data, financial information). Because this e-mail has been sent without encryption, individuals other than the intended recipient may be able to view the information, forward it to others or tamper with the information without the knowledge or consent of the sender. If you are not the intended recipient, or the employee or person responsible for delivering the message to the intended recipient, any dissemination, distribution or copying of the communication is strictly prohibited. If you received the communication in error, please notify the sender immediately by replying to this message and deleting the message and any accompanying files from your system. If, due to the security risks, you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender. --------------------------------------------------------------------- No virus found in this incoming message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: 4/02/2009 4:35 PM No virus found in this outgoing message. Checked by AVG. Version: 7.5.552 / Virus Database: 270.10.19/1938 - Release Date: 6/02/2009 5:28 PM |
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Renato A. Mortara |
Re: Input on the Leica STED
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Guy,
Thanks again for the input ! Renato Renato A. Mortara Disciplina de Parasitologia UNIFESP Escola Paulista de Medicina R. Botucatu, 862 6o andar 04023-062 S?o Paulo SP Brasil Quoting Guy Cox <[hidden email]>: > I wasn't suggesting the Leica resonant scanner wasn't good! > Leaving aside the banter about Leica's 'illegitimate' use > of the term 'tandem scanner', there is another point that's > worth making here, and that is that it's a bad idea to get > too many different features on the one stand. > > In Renato's example, a very expensive STED super-resolution > system could be at risk of standing idle for a large part of > the time while people use the resonant scanner for live-cell > experiments. This can get very frustrating, and it's hardly > sensible use of a million-dollar-plus STED instrument. Far > better to scrape together (relatively) few additional dollars > to set up a package deal on two instruments each optimised > for one task. > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) > Electron Microscope Unit, Madsen Building F09, > University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Armstrong, > Brian > Sent: Saturday, 7 February 2009 4:07 AM > To: [hidden email] > Subject: Re: Input on the Leica STED > > I have "played with" the Leica resonant scanner and I was quite > impressed. The imaging is very fast and the images are relatively > resolved given the speed. The system is also very easy to use and > allows you to average and to utilize an 8kx8K resolution which > cleans up the image considerably. However, once you perform > considerable averaging and use such a high resolution you lose the > speed. The take home message is that you have a lot of flexibility > to perform either; very fast somewhat noisy imaging, or fast and > somewhat resolved imaging, and everywhere in between. What settings > you choose depends entirely on what you want to accomplish. > Cheers, > > Brian D Armstrong PhD > Light Microscopy Core Manager > Beckman Research Institute > City of Hope > Dept of Neuroscience > 1450 E Duarte Rd > Duarte, CA 91010 > 626-256-4673 x62872 > http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Guy Cox > Sent: Friday, February 06, 2009 4:13 AM > To: [hidden email] > Subject: Re: Input on the Leica STED > > Well, maybe Moji will sue - I believe he registered a trademark! > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope > Unit, Madsen Building F09, University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of simon walker > (BI) > Sent: Friday, 6 February 2009 11:08 PM > To: [hidden email] > Subject: Re: Input on the Leica STED > > They're obviously using the term in a different context now. > From the Leica website: > > High Speed and High Resolution - All in One The Leica Tandem Scanner > combines two technological solutions in one system: a conventional > scanning system - ideal for morphological imaging and three > dimensional structure analyses, and a resonant scanning system - the > best solution for high speed confocal imaging. > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Guy Cox > Sent: 06 February 2009 12:04 > To: [hidden email] > Subject: Re: Input on the Leica STED > > Well, tandem scanning was the term coined by Mojimir Petran for his > spinning-disk confocal system. Ever since then there have been > companies offering similar systems. Leica used to offer such a > system (based on the BD CARV) but have since discontinued it. But > it is something very different in all respects (including both price > and performance) from the Leica STED which is a super- resolution > system offering 85nm resolution. > > Guy > > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope > Unit, Madsen Building F09, University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of simon walker > (BI) > Sent: Friday, 6 February 2009 10:46 PM > To: [hidden email] > Subject: Re: Input on the Leica STED > > Presumably we're talking about the SP5 with conventional and > resonant scanners and adapted for STED..? > > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Guy Cox > Sent: 06 February 2009 11:41 > To: [hidden email] > Subject: Re: Input on the Leica STED > > Hello, what are we talking about here? I've used the Leica STED > (and I'm seriously impressed with it) but I didn't know Leica made a > tandem scanner. Please clarify! I'm presenting results from the > Leica STED at NZCM next week in Rotorua and in April at FOM in > Krakow. I do realise both are a bit remote from Brazil. > > > Guy > > > Optical Imaging Techniques in Cell Biology > by Guy Cox CRC Press / Taylor & Francis > http://www.guycox.com/optical.htm > ______________________________________________ > Associate Professor Guy Cox, MA, DPhil(Oxon) Electron Microscope > Unit, Madsen Building F09, University of Sydney, NSW 2006 > ______________________________________________ > Phone +61 2 9351 3176 Fax +61 2 9351 7682 > Mobile 0413 281 861 > ______________________________________________ > http://www.guycox.net > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Renato Mortara > Sent: Friday, 6 February 2009 9:37 PM > To: [hidden email] > Subject: Input on the Leica STED > > Hi, > > I would appreciate any input on the Leica Tandem Scanner confocal > microscope (pros and cons). > > Many thanks ! > > Renato > > Renato A. Mortara > Parasitology Division > UNIFESP - Escola Paulista de Medicina > Rua Botucatu, 862, 6th floor > São Paulo, SP > 04023-062 > Brazil > Phone: 55 11 5579-8306 > Fax: 55 11 5571-1095 > email: [hidden email] > home page: www.ecb.epm.br/~ramortara > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > > --------------------------------------------------------------------- > > SECURITY/CONFIDENTIALITY WARNING: > This message and any attachments are intended solely for the > individual or entity to which they are addressed. This communication > may contain information that is privileged, confidential, or exempt > from disclosure under applicable law (e.g., personal health > information, research data, financial information). Because this > e-mail has been sent without encryption, individuals other than the > intended recipient may be able to view the information, forward it > to others or tamper with the information without the knowledge or > consent of the sender. If you are not the intended recipient, or the > employee or person responsible for delivering the message to the > intended recipient, any dissemination, distribution or copying of > the communication is strictly prohibited. If you received the > communication in error, please notify the sender immediately by > replying to this message and deleting the message and any > accompanying files from your system. If, due to the security risks, > you do not wish to receive further communications via e-mail, please > reply to this message and inform the sender that you do not wish to > receive further e-mail from the sender. > --------------------------------------------------------------------- > > No virus found in this incoming message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.18/1935 - Release Date: > 4/02/2009 4:35 PM > > > No virus found in this outgoing message. > Checked by AVG. > Version: 7.5.552 / Virus Database: 270.10.19/1938 - Release Date: > 6/02/2009 5:28 PM > > > |
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Farid Jalali |
Re: Gelatin/Lysine/Collagen/Etc.
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In reply to this post by B. Prabhakar Pandian
Poly-Lysine works quite well. I have only ever tried it myself with ex vivo dis-aggregated keratinocytes, murine and human, and murine and human lymphocytes. Simply make up the solution and use as per the instructions (Sigma works well). Suggest using it with No. 1.5 coverslips so you can plate your cells directly onto coverslips and avoid having to image to the slide or through plastic.
Best Regards Farid Jalali On Thu, Feb 5, 2009 at 8:47 AM, B. Prabhakar Pandian <[hidden email]> wrote: Hello, |
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B. Prabhakar Pandian |
Re: Gelatin/Lysine/Collagen/Etc.
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In reply to this post by Dr. Mark A. DeCoster
Thanks. I will give it a try.
-Prabhakar Mark A. DeCoster wrote: > Poly-lysine is a good one to try first and may be the easiest to work with. > As suggested cells grow better on plastic than on glass, but the optics are > better on glass. > > -MD > > Dr. Mark A. DeCoster > Associate Professor > Biomedical Engineering > Louisiana Tech University > > http://www2.latech.edu/~decoster/ > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On > Behalf Of Megan Nicholson > Sent: Friday, February 06, 2009 8:38 AM > To: [hidden email] > Subject: Re: Gelatin/Lysine/Collagen/Etc. > > Have you tried using plastic slides? Most adherent cells really grow well > on plastic. ibidi sells plastic slides with the optical quality of glass. > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On > Behalf Of B. Prabhakar Pandian > Sent: Thursday, February 05, 2009 10:48 AM > To: [hidden email] > Subject: Gelatin/Lysine/Collagen/Etc. > > Hello, > Does anybody have advice on what substrates > (Gelatin/Lysine/Collagen/Etc.) is best for growing adherent cells on > glass slides. Is there a study where cell differentiation was compared > on various substrates. > > Any help would be great. > > Thanks, > > -Prabhakar > > > -- |
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In reply to this post by Guy Cox
I second that! We have a 2-photon and confocal microscope; it's two-photon features were lying dormant to make way for a bunch of users who wanted to use the confocal function on cell cultures! We recently got a separate dedicated confocal system for them and the lab is much more productive!
Craig On Fri, Feb 6, 2009 at 6:07 PM, Guy Cox <[hidden email]> wrote: I wasn't suggesting the Leica resonant scanner wasn't good! |
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B. Prabhakar Pandian |
Multi-well plates for fluorescence studies
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In reply to this post by B. Prabhakar Pandian
Hello,
Can somebody recommend good multi-well plates for fluorescence studies. We are trying to study transfection of cells with GFP and the current plates are having the cells auto fluoresce. Thanks, -Prabhakar |
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Naomi Book |
Re: Multi-well plates for fluorescence studies
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we are using the 8 well slide from IBIDI, we use them with 60X oil immersion
objective and get very nice results. (no commercial interest) Naomi Book Dr. Naomi Melamed-Book Bio-Imaging Unit Life Science Institute Hebrew University, Givaat Ram, Jerusalem, Israel phone: 972-2-6585453 ----- Original Message ----- From: "B. Prabhakar Pandian" <[hidden email]> To: <[hidden email]> Sent: Thursday, March 05, 2009 11:20 PM Subject: Multi-well plates for fluorescence studies > Hello, > Can somebody recommend good multi-well plates for fluorescence > studies. We are trying to study transfection of cells with GFP and the > current > plates are having the cells auto fluoresce. > > Thanks, > > -Prabhakar > > > -- > No virus found in this incoming message. > Checked by AVG. Version: 7.5.557 / Virus Database: 270.11.8/1985 - Release > Date: 5/3/2009 07:54 > > |
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Csúcs Gábor |
Re: Multi-well plates for fluorescence studies
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In reply to this post by B. Prabhakar Pandian
Dear Prabhakar,
You haven't indicated whether you need 96 or 384 well plates. Generally glass bottom plates offer very good image quality: You can get then from BD or from Porvair (these are the cheapest I know), but there are certainly other vendors as well. If you want thin-plastic bottom plates, then we have good experiences with Matrix and Greiner, but also with BD plates (given that you buy an imaging plate). But again there are other vendors as well. If you need only 8 wells, the the mentioned Ibidi or the LabtTek slides (with cover-slip bottom) are a good choice. Greetings Gabor P.S. No commercial interest. |
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Keith Morris |
Re: multiwell plates/slides/petri dishes for high mag fluorescence imaging studies
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In reply to this post by Naomi Book
Mattek make a wide selection of multiwell plates [and Petri dishes] with
cover-slip bases for inverted live/fixed cell imaging: http://www.glassbottomdishes.com. You can even drop vectashield or similar [non-hardset] antifadent into the well after fixing/staining, and this setup stores well in the fridge between imaging. Also see link below. I do try to keep a list of culture vessels that can be used for high magnification imaging with an inverted fluorescence microscope, e.g. Mattek Petri dishes, Mattek Multiwell plates, CultureWell & LabTek 3x1" slides, all of which can be supplied in versions with a cover-slip base for high mag imagimg with high NA oil objectives. Mattek Petri dishes can even come with a grid etched on the cover-slip for relocating cells or special coatings on the coverslip for fussy cells. The culture vessels we use for imaging are linked via our 'external microscopy sites' on our website. If you know of others that are available [and great to use] I would be happy to hear about them off-line so that the links could be updated: http://www.well.ox.ac.uk/cytogenetics/websites.shtml See: Microscopy Live Cell Imaging and Anti-Fadent Mountants Nowhere near the standard of sites like TAVI's FISH pages, but hopefully of use to some. Keith --------------------------------------------------------------------------- Dr Keith J. Morris, Molecular Cytogenetics and Microscopy Core, Laboratory 00/069 and 00/070, The Wellcome Trust Centre for Human Genetics, Roosevelt Drive, Oxford OX3 7BN, United Kingdom. Telephone: +44 (0)1865 287568 Email: [hidden email] Web-pages: http://www.well.ox.ac.uk/cytogenetics/ -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Naomi Book Sent: 05 March 2009 21:43 To: [hidden email] Subject: Re: Multi-well plates for fluorescence studies we are using the 8 well slide from IBIDI, we use them with 60X oil immersion objective and get very nice results. (no commercial interest) Naomi Book Dr. Naomi Melamed-Book Bio-Imaging Unit Life Science Institute Hebrew University, Givaat Ram, Jerusalem, Israel phone: 972-2-6585453 ----- Original Message ----- From: "B. Prabhakar Pandian" <[hidden email]> To: <[hidden email]> Sent: Thursday, March 05, 2009 11:20 PM Subject: Multi-well plates for fluorescence studies > Hello, > Can somebody recommend good multi-well plates for fluorescence > studies. We are trying to study transfection of cells with GFP and the > current > plates are having the cells auto fluoresce. > > Thanks, > > -Prabhakar > > > -- > No virus found in this incoming message. > Checked by AVG. Version: 7.5.557 / Virus Database: 270.11.8/1985 - Release > Date: 5/3/2009 07:54 > > |
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