Biopsy imaging/holding tools
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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, In the process of imaging small (2-4 mm) tissue biopsies under a confocal microscope with a water-immersion objective, we have had problems holding the tissue sample down to avoid flotation in water. We have tried using tweezers with a clip to hold them but are afraid that may damage the biopsy in some instances, and is not always possible for irregular shaped samples. Can anyone provide suggestions as to how this problem can be solved either with a commercial or a custom method? Thanks so much, Bilal Malik |
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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 Bilal; Here's what we use... https://www.warneronline.com/product_info.cfm?id=80&alpha=a&name=Slice%20Anchor%20Kits%20and%20Hold-Downs%20%28Harps%29 Old-school types use nylon stockings pulled over a small metal frame, then pull out selected fibers until you get the density you need to hold down your sample. Kathy Kathryn R. Spencer, PhD The Scripps Research Institute 10550 N. Torrey Pines Road, DNC 210 La Jolla, CA 92037 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Bilal Malik Sent: Wednesday, April 10, 2013 9:38 AM To: [hidden email] Subject: Biopsy imaging/holding tools ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear all, In the process of imaging small (2-4 mm) tissue biopsies under a confocal microscope with a water-immersion objective, we have had problems holding the tissue sample down to avoid flotation in water. We have tried using tweezers with a clip to hold them but are afraid that may damage the biopsy in some instances, and is not always possible for irregular shaped samples. Can anyone provide suggestions as to how this problem can be solved either with a commercial or a custom method? Thanks so much, Bilal Malik |
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In reply to this post by Bilal Malik
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Bilal, Have you considered using cyanoacrylate to glue the tissue to the dish? There are cyanoacrylate based gels used to hold tissues together after surgery so they should be safe for your biopsies. Chris Tully, M.S., Image Analysis Expert t 240.475.9753 f 419.831.0527 | [hidden email] Sent from my iPhone please excuse typos. On Apr 10, 2013, at 12:49 PM, Bilal Malik <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear all, > > > > In the process of imaging small (2-4 mm) tissue biopsies under a confocal > microscope with a water-immersion objective, we have had problems holding > the tissue sample down to avoid flotation in water. We have tried using > tweezers with a clip to hold them but are afraid that may damage the biopsy > in some instances, and is not always possible for irregular shaped samples. > Can anyone provide suggestions as to how this problem can be solved either > with a commercial or a custom method? > > > > Thanks so much, > > Bilal Malik > > |
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In reply to this post by Bilal Malik
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** When imaging brain slices, we commonly use a so-called grid: a piece of platinum wire is flattened and bent into a U shape large enough to easily surround your sample. Next very thin threads from nylon stockings (the single filaments!) are glued onto this frame, taking care that the glue is on the outer sides of the frame, so that the frame and the filaments themselves can rest on the bottom of the imaging chamber. Also make sure that the threads are under tension. This grid (or "harp") gets placed on top of your sample. Obviously, if the sample is fairly thick ( > 300 microns), you might want to have less tension on the filaments. Some people also use small (5 - 10 mm long) pieces of silver or platinum wire to weigh down the sample. Hope that helps, Chris > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear all, > > > > In the process of imaging small (2-4 mm) tissue biopsies under a confocal > microscope with a water-immersion objective, we have had problems holding > the tissue sample down to avoid flotation in water. We have tried using > tweezers with a clip to hold them but are afraid that may damage the biopsy > in some instances, and is not always possible for irregular shaped samples. > Can anyone provide suggestions as to how this problem can be solved either > with a commercial or a custom method? > > > > Thanks so much, > > Bilal Malik > > > |
 
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Glen MacDonald-2 |
Re: Biopsy imaging/holding tools
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In reply to this post by Bilal Malik
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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 Bilal, Harps work well, although you have to focus between the threads. If building your own, nylon threads are a little fluorescent, but usually not a problem with confocal. Cyanoacrylate glue can also be fluorescent, but careful placement can avoid issues. Low melting point agarose is also useful. Use at 1% to 3% concentration, depending on desired hardness and density. If the samples are fixed, then harps can be made from stainless steel wire rather than more expensive metals. I found a good source of stainless wire at a sporting goods store which sold wire for building fishing lures. Regards, Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Cellular Morphology Core Center on Human Development and Disability Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 [hidden email] On Apr 10, 2013, at 9:37 AM, Bilal Malik <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear all, > > > > In the process of imaging small (2-4 mm) tissue biopsies under a confocal > microscope with a water-immersion objective, we have had problems holding > the tissue sample down to avoid flotation in water. We have tried using > tweezers with a clip to hold them but are afraid that may damage the biopsy > in some instances, and is not always possible for irregular shaped samples. > Can anyone provide suggestions as to how this problem can be solved either > with a commercial or a custom method? > > > > Thanks so much, > > Bilal Malik > > |
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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 all so much for your replies. One thing I failed to mention is that biopsies are fresh tissue, and so all the imaging needs to be done within a relatively short time (10-20 minutes) in order to avoid degradation. Afterwards the tissue is fixed. Thanks, Bilal -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Glen MacDonald Sent: Wednesday, April 10, 2013 1:03 PM To: [hidden email] Subject: Re: Biopsy imaging/holding tools ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Bilal, Harps work well, although you have to focus between the threads. If building your own, nylon threads are a little fluorescent, but usually not a problem with confocal. Cyanoacrylate glue can also be fluorescent, but careful placement can avoid issues. Low melting point agarose is also useful. Use at 1% to 3% concentration, depending on desired hardness and density. If the samples are fixed, then harps can be made from stainless steel wire rather than more expensive metals. I found a good source of stainless wire at a sporting goods store which sold wire for building fishing lures. Regards, Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Cellular Morphology Core Center on Human Development and Disability Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 [hidden email] On Apr 10, 2013, at 9:37 AM, Bilal Malik <[hidden email]> wrote: > ***** > To join, leave or search the confocal microscopy listserv, go to: > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy > ***** > > Dear all, > > > > In the process of imaging small (2-4 mm) tissue biopsies under a > confocal microscope with a water-immersion objective, we have had > problems holding the tissue sample down to avoid flotation in water. > We have tried using tweezers with a clip to hold them but are afraid > that may damage the biopsy in some instances, and is not always possible > Can anyone provide suggestions as to how this problem can be solved > either with a commercial or a custom method? > > > > Thanks so much, > > Bilal Malik > > |
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In reply to this post by Bilal Malik
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To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Bilal Here is a link to the use of a Corning Snapwell™ turned upside down for upright scopes or right side up with the use of a glass outer ring. http://www.bioptechs.com/Products/Delta_T/Options/options.html#snapwell This is typically used for keeping a specimen in contact with a membrane while imaging on a upright scope. Some people have cut a hole or slit in the membrane to secure their specimen within the tension of the membrane. You get six of Snapwell membranes in a SBS format plate. You simply pop off the support holder of the membrane and throw it away and just use the shallow membrane fixture. The Delta T dish 23mm ID is ideal for the footprint of the Snapwell and fits snugly into a stage adapter. If you need temperature control the dish provides warmth through a closed loop feedback system. (See Delta T product info page) Other options: These links are to a Z axis controlled accessory for the Delta T dish that supports either a coverslip or a membrane immediately above the specimen for inverted microscopes. The membrane suspension device is used when you need maximum media contact to the specimen and the ability to keep the specimen in place against the coverslip. http://www.bioptechs.com/Products/Delta_T/Options/options.html#membraneadapter Here is a similar technique that uses a coverslip instead of a membrane. http://www.bioptechs.com/Products/Delta_T/Options/options.html#mediadepth Here is a link to a product we call a brain slice adapter although it can be used any tissue. Your specimen is placed on a polycarbonate disk that is lowered into a Delta T Dish with the same type Z axis translator as the above devices where its proximity to the coverslip bottom dish is adjustable. There is a nylon weight that rest on the top of the specimen to prevent it from floating. http://www.bioptechs.com/Products/Delta_T/Options/options.html#Brain%20A Dan On Apr 10, 2013, at 12:37 PM, Bilal Malik wrote: ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear all, In the process of imaging small (2-4 mm) tissue biopsies under a confocal microscope with a water-immersion objective, we have had problems holding the tissue sample down to avoid flotation in water. We have tried using tweezers with a clip to hold them but are afraid that may damage the biopsy in some instances, and is not always possible for irregular shaped samples. Can anyone provide suggestions as to how this problem can be solved either with a commercial or a custom method? Thanks so much, Bilal Malik 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 Bilal Malik
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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 Bilal, I've used a few basic solutions in the past, anything from a simple metal washer to super-gluing the sample to the bottom of a small petri-dish (assuming the sample isn't precious and recovery required). Another slightly more technical solution would be to use a product called Cy-gel (no commercial interest - just what we have here in the facility), which basically is liquid in the fridge and sets like agarose when it warms to room-temperature. Failing getting hold of that you could use some low melting point agarose instead but it will depend on how sensitive your sample is to heat for that to be applicable. Hope that helps, Mark Scott FILM - Facility for Imaging by Light Microscopy Senior Research Technician Sir Alexander Fleming Building, desk 408 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel: ++44(0)20-759-49793 E-mail: [hidden email] Website: http://imperial.ac.uk/imagingfacility -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Bilal Malik Sent: 10 April 2013 17:38 To: [hidden email] Subject: Biopsy imaging/holding tools ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear all, In the process of imaging small (2-4 mm) tissue biopsies under a confocal microscope with a water-immersion objective, we have had problems holding the tissue sample down to avoid flotation in water. We have tried using tweezers with a clip to hold them but are afraid that may damage the biopsy in some instances, and is not always possible for irregular shaped samples. Can anyone provide suggestions as to how this problem can be solved either with a commercial or a custom method? Thanks so much, Bilal Malik |
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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, Thank you very much for your responses. I had no idea that these harp-like devices exist - those would definitely be helpful. Also, using Cy-gel and agarose are very helpful ideas. Thanks again, Bilal -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Scott, Mark Sent: Thursday, April 11, 2013 4:23 AM To: [hidden email] Subject: Re: Biopsy imaging/holding tools ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear Bilal, I've used a few basic solutions in the past, anything from a simple metal washer to super-gluing the sample to the bottom of a small petri-dish (assuming the sample isn't precious and recovery required). Another slightly more technical solution would be to use a product called Cy-gel (no commercial interest - just what we have here in the facility), which basically is liquid in the fridge and sets like agarose when it warms to room-temperature. Failing getting hold of that you could use some low melting point agarose instead but it will depend on how sensitive your sample is to heat for that to be applicable. Hope that helps, Mark Scott FILM - Facility for Imaging by Light Microscopy Senior Research Technician Sir Alexander Fleming Building, desk 408 Imperial College London / South Kensington Exhibition Road London SW7 2AZ UK Tel: ++44(0)20-759-49793 E-mail: [hidden email] Website: http://imperial.ac.uk/imagingfacility -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Bilal Malik Sent: 10 April 2013 17:38 To: [hidden email] Subject: Biopsy imaging/holding tools ***** To join, leave or search the confocal microscopy listserv, go to: http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy ***** Dear all, In the process of imaging small (2-4 mm) tissue biopsies under a confocal microscope with a water-immersion objective, we have had problems holding the tissue sample down to avoid flotation in water. We have tried using tweezers with a clip to hold them but are afraid that may damage the biopsy in some instances, and is not always possible for irregular shaped samples. Can anyone provide suggestions as to how this problem can be solved either with a commercial or a custom method? Thanks so much, Bilal Malik |
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