Near IR Imaging
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Does anyone have suggestions on how to set up a microscope for imaging Near IR dyes? We would like to image Cy7, Cy7.5 and VisTag S750. We have Zeiss 510, 710 microscopes as well as Olympus FV1000 and TIRF3 microscopes. Thanks for your help. Kim Peifley Kim Peifley (Contractor) Research Associate I Optial Microscopy and Analysis Laboratory http://atp.ncifcrf.gov SAIC-Frederick, Inc. National Cancer Institute at Frederick Post Office Box B Frederick, MD 21702 Phone: 301-846-6561 Fax: 301-846-7672 [hidden email] |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Kim, please fill free to adopt the MIAWiki webpage: http://confocal-manawatu.pbworks.com/w/page/16346945/Leica%20Confocal%20SP5%20Quick%20Starter%20Guide as a template for manuals for your instruments and store your work at MIAWiki for your quick reference in future. References to the principles of fluorescence microscopy and guides on selection of fluorescence cubes are made accessible in seconds through the MIAWiki Search bar. Please fill free to modify the content and structure according to your vision of how it should be done. With kind regards, Dmitry Sokolov MIAWiki for Mass Collaboration: Share or Loose :0) On 9/12/2011 10:05 a.m., Kim Peifley wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Does anyone have suggestions on how to set up a microscope for imaging > Near IR dyes? We would like to image Cy7, Cy7.5 and VisTag S750. We have > Zeiss 510, 710 microscopes as well as Olympus FV1000 and TIRF3 microscopes. > > Thanks for your help. > > Kim Peifley > > > Kim Peifley (Contractor) > Research Associate I > Optial Microscopy and Analysis Laboratory > http://atp.ncifcrf.gov > SAIC-Frederick, Inc. > National Cancer Institute at Frederick > Post Office Box B > Frederick, MD 21702 > Phone: 301-846-6561 > Fax: 301-846-7672 > [hidden email] > |
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In reply to this post by Kim Peifley
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Kim, First, i think, you have to change your primary dichroic miror and use the 760nm. Do you have an idea of the emission peak ? May be you can use an OPO between your pulsed laser and your microscope. If you have enough money you have this solution: INSIGTH by SpectraPhysics. regards |
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In reply to this post by Dmitry Sokolov
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
 
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Sylvie Le Guyader-2 |
Re: Dishes for live cell imaging
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Hi Julia One of our users successfully grows rat primary neurons on MatTek 35mm glass bottom dishes coated w Poly-L-Lysine. Coating with laminin (working concentration 10 ug/ml) usually helps. If you are after axons, they sometimes take several days to regrow after being cut. Med vänlig hälsning / Best regards Sylvie @@@@@@@@@@@@@@@@@@@@@@@@ Sylvie Le Guyader Live Cell Imaging Unit Dept of Biosciences and Nutrition Karolinska Institutet Novum 14183 Huddinge Sweden office: +46 (0) 8 5248 1107 LCI room: +46 (0) 8 5248 1172 mobile: +46 (0) 73 733 5008 > -----Original Message----- > From: Confocal Microscopy List > [mailto:[hidden email]] On Behalf Of Julia Edgar > Sent: 09 December 2011 09:08 > To: [hidden email] > Subject: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear All > I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes > with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have > had variable degrees of success so far, mainly because the primary neuronal cells > do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). > > I will be grateful to hear you recommendations for dishes and/or substrate. > > Thank you in advance. > Julia Edgar > University of Glasgow > |
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In reply to this post by Julia Edgar
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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tineke vendrig |
Re: Dishes for live cell imaging
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We also made them ourself, while I was working at the VU in Amsterdam. Just saw a hole in the dish end glue a coverglass on/in it. Then you can coat the glass and culture your material! Easy as you always do! Tineke Vendrig 2011/12/9 MODEL, MICHAEL <[hidden email]> > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Those are quite easy to make in the lab. We make them all the time and in > large quantities. If interested, I can send the instructions. > > Mike Model > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On > Behalf Of Julia Edgar [[hidden email]] > Sent: Friday, December 09, 2011 3:07 AM > To: [hidden email] > Subject: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear All > I need to purchase glass-bottomed dishes (currently we use 35 mm Petri > dishes with ~15 mm diameter glass bottom) for live-imaging of primary > neural cells. I have had variable degrees of success so far, mainly because > the primary neuronal cells do not adhere well to the dishes (which were > coated with 0.01% poly-L-lysine). > > I will be grateful to hear you recommendations for dishes and/or substrate. > > Thank you in advance. > Julia Edgar > University of Glasgow > -- Tineke Vendrig, ing technical engineer optical microscopy Delft University of Technology Bionano Science Kavli Institute of Nanoscience Lorentzweg 1 2628LJ Delft room F185 Tel: +31 15 2789299 Fax:+31 15 2781202 email: [hidden email] mobile phone: +31 624341412 |
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In reply to this post by Kim Peifley
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We are trying to image ICG dye (indocyanine green, Ex. ~ 800nm) and getting a filter cube was simple. The difficulty is finding a light source that emits at a high enough frequency and making sure the detector, in our case, a CCD camera can see it. Manufacturers often put IR filters in both the camera and light source against potential heat damage so those have to be removed. I also found information about light source output from companies to be inconsistent as if they don't have too much experience with this range of wavelengths. Currently, we are dusting off an olde Xenon light source and will take a look. Judy Judy Trogadis Bio-Imaging Coordinator Keenan Research Centre, St. Michael's 209 Victoria Street Toronto M5B 1T8, Canada office: 416-864-6060 ext. 77612 imaging facility: ext. 77434 cell: 416-909-9878 [hidden email] -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kim Peifley Sent: Thursday, December 08, 2011 4:06 PM To: [hidden email] Subject: Near IR Imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Does anyone have suggestions on how to set up a microscope for imaging Near IR dyes? We would like to image Cy7, Cy7.5 and VisTag S750. We have Zeiss 510, 710 microscopes as well as Olympus FV1000 and TIRF3 microscopes. Thanks for your help. Kim Peifley Kim Peifley (Contractor) Research Associate I Optial Microscopy and Analysis Laboratory http://atp.ncifcrf.gov SAIC-Frederick, Inc. National Cancer Institute at Frederick Post Office Box B Frederick, MD 21702 Phone: 301-846-6561 Fax: 301-846-7672 [hidden email] |
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In reply to this post by mmodel
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Thank you, Mike. Would be great to have the instructions. Best wishes Julia ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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Angela V Klaus |
Re: Dishes for live cell imaging
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I would love to have the instructions as well. Many thanks, Angela Angela V. Klaus, PhD Department of Biological Sciences Seton Hall University ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 10:19 AM To: [hidden email] Subject: Re: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Thank you, Mike. Would be great to have the instructions. Best wishes Julia ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I am receiving many requests for instruction for making dishes, maybe it would make sense to post them. This is how we do it: You would need: 1. Roper Whitney XX hand punch (Roper Whitney #135010001 with bench mounting base #139010001) to make holes in the bottom of plastic dishes. 2. Hand deburring tool (Small Parts Inc #DBR-HT) 3. Sylgard 184 glue (World Precision Instruments, #SYLG184) to glue glass coverslips to dishes. It cures overnight and is nontoxic to cells. (From: D. Kline in Microinjections: Methods and applications, ed. DJ Carroll, 2009 Humana Press, pp 135-156) -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Angela V Klaus Sent: Friday, December 09, 2011 10:30 AM To: [hidden email] Subject: Re: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** I would love to have the instructions as well. Many thanks, Angela Angela V. Klaus, PhD Department of Biological Sciences Seton Hall University ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 10:19 AM To: [hidden email] Subject: Re: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Thank you, Mike. Would be great to have the instructions. Best wishes Julia ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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In reply to this post by Kim Peifley
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We are routinely imaging Cy7 and IRDye 800 using a xenon lamp and a backthinned CCD (Hamamatsu Orca BT 1024 G) on a widefield scope. It works fine for reasonably bright signals, especially immunofluorescence. We bought the Cy7 filter set from Chroma. We removed all the heat blocking filters from the light path. You might do better with a far red LED light source but we haven't tried this. The LICOR instruments (not microscopes) apparently use far red laser excitation and APDs to detect. We have found that signals can be detected on their tissue scanner that we cannot see with our microscope set up. Without an OPO you probably can't do this on a multiphoton system and in any case the detectors on most confocals fall off steeply in the near IR. There is a group at Rice that is developing a widefield system using InGaAs cameras for detection: http://pubs.acs.org/doi/abs/10.1021/ja0466311 |
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Donnelly, Tom |
Re: Dishes for live cell imaging
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In reply to this post by Julia Edgar
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Julia; There are several vendors of glass bottom 35mm dishes. It is strange but true that cells like some glasses better than others. You may want to try WillcoWells or Ibidi. Ibidi actually uses plastic bottoms that have the optical properties of a #1.5 glass coverslip. Using DIY and/or commercial dishes one thing to keep in mind is the cover slip needs to be orthogonal to the light path. A slanted coverslip is going to create optical issues in your image. Cheers, Tom -----Original Message----- From: Julia Edgar [mailto:[hidden email]] Sent: Friday, December 09, 2011 12:08 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow This email message, together with any attachments, is for the sole use of the intended recipient(s) and is the confidential information of Applied Precision Inc. If you are not the intended recipient, your review, use, disclosure, copying or dissemination of this email message or its attachments, or the information contained therein, is strictly prohibited. If you are not the intended recipient or if you think this email was sent to you in error, please notify the sender by reply email and delete this message and its attachments, as well as all copies, from your system. |
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In reply to this post by mmodel
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We use 25mm, #1.5, round coverslips with a stainless steel, two-piece, reusable dish from invitrogen (Atofluor Cell Chamber A7816). The cost for the chamber is $280 for three, http://probes.invitrogen.com/media/pis/mp07816.pdf Eric Marino [hidden email] [hidden email] Cell: (617) 913-9647 Lab: (617) 713-8885 Sent from my mobile device On Dec 9, 2011, at 11:05 AM, "MODEL, MICHAEL" <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I am receiving many requests for instruction for making dishes, maybe it would make sense to post them. This is how we do it: > > You would need: > > 1. Roper Whitney XX hand punch (Roper Whitney #135010001 with bench mounting base #139010001) to make holes in the bottom of plastic dishes. > 2. Hand deburring tool (Small Parts Inc #DBR-HT) > 3. Sylgard 184 glue (World Precision Instruments, #SYLG184) to glue glass coverslips to dishes. It cures overnight and is nontoxic to cells. > > (From: D. Kline in Microinjections: Methods and applications, ed. DJ Carroll, 2009 Humana Press, pp 135-156) > > > > > > -----Original Message----- > From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Angela V Klaus > Sent: Friday, December 09, 2011 10:30 AM > To: [hidden email] > Subject: Re: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > I would love to have the instructions as well. Many thanks, Angela > > Angela V. Klaus, PhD > Department of Biological Sciences > Seton Hall University > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] > Sent: Friday, December 09, 2011 10:19 AM > To: [hidden email] > Subject: Re: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > > > Thank you, Mike. Would be great to have the instructions. > > Best wishes > > Julia > > > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> > ***** > > Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. > > Mike Model > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] > Sent: Friday, December 09, 2011 3:07 AM > To: [hidden email] > Subject: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy <http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> > ***** > > Dear All > I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). > > I will be grateful to hear you recommendations for dishes and/or substrate. > > Thank you in advance. > Julia Edgar > University of Glasgow >
Eric Marino
Senior Imaging Specialist Program in Cellular and Molecular Medicine Boston Children's Hospital |
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In reply to this post by Kim Peifley
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** That far in the NIR do not be surprised if none of your microscopes are parfocal with visible wavelengths. You could try your LSM710 and FV1000 with your current laser lines - especially 633/642 nm, though the PMT(s) have low quantum efficiency in the NIR. I suggest you build up from Cy5 / Alexa Fluor 647, Cy5.5, Alexa Fluor 680 and other NIF Alexa Fluor's, and on up to longer wavelengths. See also perkinElmer/Caliper/VisEn Medical's 680 reagents (for examples, http://www.perkinelmer.com/Catalog/Category/ID/Activatable ) as alternatives to the 750 series. For example, if you were to use droplets of (say) 1 uM concentration of each dye, if you can see Cy5 but not Cy5.5, I doubt you will see Cy7 etc. You should be able to spot the edge of the droplet by eye with trasmitted light, and all of these dyes have some absorption at 633/642 nm, so you should be able to visualize them with the transmitted light detector. You can also try opening the confocal pinhole. Your TIRF3 microscope is probably the best bet. If the digital camera has an IR blocking filter (see replies by others), you may not get any signal. You should also ask your scope vendors for help. best wishes, George On 12/8/2011 4:05 PM, Kim Peifley wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Does anyone have suggestions on how to set up a microscope for imaging > Near IR dyes? We would like to image Cy7, Cy7.5 and VisTag S750. We have > Zeiss 510, 710 microscopes as well as Olympus FV1000 and TIRF3 microscopes. > > Thanks for your help. > > Kim Peifley > > > Kim Peifley (Contractor) > Research Associate I > Optial Microscopy and Analysis Laboratory > http://atp.ncifcrf.gov > SAIC-Frederick, Inc. > National Cancer Institute at Frederick > Post Office Box B > Frederick, MD 21702 > Phone: 301-846-6561 > Fax: 301-846-7672 > [hidden email] > > -- George McNamara, PhD Analytical Imaging Core Facility University of Miami |
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In reply to this post by Judy Trogadis-2
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** We have a Zeiss 710 with three spectral detectors. Detection is fine up to 740 nm. Between 740 and 750 nm the instensity falls off quickly. Nothing is detectable above 750 nm. We measured this by reflection of a MaiTai laser. _________________________________________ Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine Lab: (212) 263-3208 Cell: (914) 309-3270 ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Judy Trogadis [[hidden email]] Sent: Friday, December 09, 2011 8:34 AM To: [hidden email] Subject: Re: Near IR Imaging We are trying to image ICG dye (indocyanine green, Ex. ~ 800nm) and getting a filter cube was simple. The difficulty is finding a light source that emits at a high enough frequency and making sure the detector, in our case, a CCD camera can see it. Manufacturers often put IR filters in both the camera and light source against potential heat damage so those have to be removed. I also found information about light source output from companies to be inconsistent as if they don't have too much experience with this range of wavelengths. Currently, we are dusting off an olde Xenon light source and will take a look. Judy Judy Trogadis Bio-Imaging Coordinator Keenan Research Centre, St. Michael's 209 Victoria Street Toronto M5B 1T8, Canada office: 416-864-6060 ext. 77612 imaging facility: ext. 77434 cell: 416-909-9878 [hidden email] -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kim Peifley Sent: Thursday, December 08, 2011 4:06 PM To: [hidden email] Subject: Near IR Imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Does anyone have suggestions on how to set up a microscope for imaging Near IR dyes? We would like to image Cy7, Cy7.5 and VisTag S750. We have Zeiss 510, 710 microscopes as well as Olympus FV1000 and TIRF3 microscopes. Thanks for your help. Kim Peifley Kim Peifley (Contractor) Research Associate I Optial Microscopy and Analysis Laboratory http://atp.ncifcrf.gov SAIC-Frederick, Inc. National Cancer Institute at Frederick Post Office Box B Frederick, MD 21702 Phone: 301-846-6561 Fax: 301-846-7672 [hidden email] ------------------------------------------------------------ This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain information that is proprietary, confidential, and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure, or distribution is prohibited. If you have received this email in error please notify the sender by return email and delete the original message. Please note, the recipient should check this email and any attachments for the presence of viruses. The organization accepts no liability for any damage caused by any virus transmitted by this email. ================================= |
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In reply to this post by Julia Edgar
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Julia Bioptechs has several products made specifically for your concerns. One is a product called the interchangeable Coverslip Dish (ICD). It has many advantages over pre-made glass bottomed dishes. No drilling, glueing, biocompatibility issues, deburring, or dealing with small apertures. Separate coverglass enables the user to pretreat or condition the coverslip if necessary Important for neurons! No metal components have contact with the specimen or media 25mm clear aperture for more X,Y translation room with todays large high N.A. objectives Accessories available for fluid perfusion Gas and condensation control or 2nd optical surface All parts are autoclaveable. Therefore, you can control sterility #1.5, 30mm Diameter German Desag glass, ideal for cell culture O-ring sealed, gentle on the coverslip and no leakage Low cost coverslip replacement Fits any 35mm heated stage insert. However, to eliminate Z axis drift due to peripheral heating we recommend the Bioptechs Stable Z warmer at the bottom of the following link. Additional Information: http://www.bioptechs.com/Products/ICD/coverslipdish.html To help your neurons plate faster see our Culture Cylinders http://www.bioptechs.com/Products/Delta_T/Options/options.html#Culture%20Cylinders The other dish product is our Delta T system. You can learn about its many attributes at: http://www.bioptechs.com/Products/Delta_T/delta_t.html Dan On Dec 9, 2011, at 3:07 AM, Julia Edgar wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow Dan Focht Bioptechs, Inc. 3560 Beck Rd. Butler, PA 16002 www.bioptechs.com P: (724)282-7145 F: (724)282-0745 [hidden email] |
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In reply to this post by mmodel
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Mike and Others who make their own dishes for live cell imaging. Any recommendations on sterilisation - a means that will not affect the properties of the glue or the plastic Petri-dish? Best wishes Julia Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Julia - I simple spray them with 70% ethanol, then rinse. Others keep dishes in a UV hood overnight. ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Tuesday, January 17, 2012 5:18 AM To: [hidden email] Subject: Re: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Mike and Others who make their own dishes for live cell imaging. Any recommendations on sterilisation - a means that will not affect the properties of the glue or the plastic Petri-dish? Best wishes Julia Those are quite easy to make in the lab. We make them all the time and in large quantities. If interested, I can send the instructions. Mike Model ________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of Julia Edgar [[hidden email]] Sent: Friday, December 09, 2011 3:07 AM To: [hidden email] Subject: Dishes for live cell imaging ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear All I need to purchase glass-bottomed dishes (currently we use 35 mm Petri dishes with ~15 mm diameter glass bottom) for live-imaging of primary neural cells. I have had variable degrees of success so far, mainly because the primary neuronal cells do not adhere well to the dishes (which were coated with 0.01% poly-L-lysine). I will be grateful to hear you recommendations for dishes and/or substrate. Thank you in advance. Julia Edgar University of Glasgow |
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In reply to this post by Julia Edgar
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Julia Here is a suggestion that is hard to beat. Bioptechs makes a special Interchangeable Coverslip Dish (ICD) with the following attributes. 25mm clear aperture for more translation room with todays large high N.A. objectives Accessories available for fluid perfusion, Heated Lid or 2nd optical surface No metal components have contact with the specimen or media Threaded tube can be made any height for special applications All parts are autoclaveable. Therefore, you can control sterility Significantly reduced Z axis drift as compared to plastic Metal base transfers heat from peripheral warmers 30mm German Desag glass ideal for cell culture Si O-ring sealed to prevent leaks Low cost coverslip replacement Fits any 35mm dish warmer, Although the Stable Z is best. See info at bottom of page. Additional Info available at http://www.bioptechs.com/Products/ICD/ coverslipdish.html As for Neuronal adherence, this glass was used in a set of images in Science showing 4 days of continual and progressive growth. If you are interested I get you the reference and images. Dan On Jan 17, 2012, at 5:18 AM, Julia Edgar wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear Mike and Others who make their own dishes for live cell imaging. > Any recommendations on sterilisation - a means that will not affect > the properties of the glue or the plastic Petri-dish? > > Best wishes > Julia > > Those are quite easy to make in the lab. We make them all the time > and in large quantities. If interested, I can send the instructions. > > Mike Model > > ________________________________________ > From: Confocal Microscopy List [[hidden email]] > On Behalf Of Julia Edgar [[hidden email]] > Sent: Friday, December 09, 2011 3:07 AM > To: [hidden email] > Subject: Dishes for live cell imaging > > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear All > I need to purchase glass-bottomed dishes (currently we use 35 mm > Petri dishes with ~15 mm diameter glass bottom) for live-imaging of > primary neural cells. I have had variable degrees of success so > far, mainly because the primary neuronal cells do not adhere well > to the dishes (which were coated with 0.01% poly-L-lysine). > > I will be grateful to hear you recommendations for dishes and/or > substrate. > > Thank you in advance. > Julia Edgar > University of Glasgow Dan Focht [hidden email] Bioptechs, Inc. 3560 Beck Rd. Butler, PA 16002 www.bioptechs.com Direct 724-282-7145 Fax 724-282-0745 Toll Free 877 LIVE-CELL - (548-3235) |
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