home-made glass-bottomed cell culture dishes
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Melinda Larsen |
home-made glass-bottomed cell culture dishes
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I have been ordering custom-made glass-bottomed culture dishes from
MatTek, but they have changed their manufacturing process such that it is fully automated and is no longer amenable to customization. Does anyone make their own glass-bottomed dishes by punching holes in the bottom of plastic culture dishes and gluing coverslips on the bottom? If so, could you give me some tips on how you do it? Thanks! Melinda Larsen, Ph.D., Assistant Professor University at Albany, SUNY Department of Biological Sciences |
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Paul Herzmark |
Re: home-made glass-bottomed cell culture dishes
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You can make gazillions easily.
Drill a big hole in the plastic dish. If you have a drill press it is assembly line speed. Circle the hole with some silicon glue (sparingly) like is used for aquariums. stick on your coverslip. Make sure it is nice a parallel to the rest of the plastic dish bottom so the whole microscope field is in focus at the same time. You can sterilize them with EtOH. Paul Herzmark Specialist [hidden email] Department of Molecular and Cell Biology 479 Life Science Addition University of California, Berkeley Berkeley, CA 94720-3200 (510) 643-9603 (510) 643-9500 fax On Wed, Jan 7, 2009 at 6:49 PM, Melinda Larsen <[hidden email]> wrote: I have been ordering custom-made glass-bottomed culture dishes from |
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Knecht, David |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Melinda Larsen
We used to do this (now using Willco Wells). We found it easier to use Glass Petri dishes as they could be easily cut by someone who knows how to drill glass (our shop did this). Then the coverslips were glued onto the bottom forming a small well in the bottom of the dish. I don't remember what glue we used but it was not hard to reglue broken ones. Dave
On Jan 7, 2009, at 9:49 PM, Melinda Larsen wrote:
Dr. David Knecht Department of Molecular and Cell Biology Co-head Flow Cytometry and Confocal Microscopy Facility U-3125 91 N. Eagleville Rd. University of Connecticut Storrs, CT 06269 860-486-2200 860-486-4331 (fax) |
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Melinda Larsen |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Paul Herzmark
Thanks, Paul! I don't have a drill press, but I'll see if my machine
shop does. If not, I'll get one. On Wed, Jan 7, 2009 at 10:50 PM, Paul Herzmark <[hidden email]> wrote: > You can make gazillions easily. > > Drill a big hole in the plastic dish. If you have a drill press it is > assembly line speed. Circle the hole with some silicon glue (sparingly) like > is used for aquariums. stick on your coverslip. Make sure it is nice a > parallel to the rest of the plastic dish bottom so the whole microscope > field is in focus at the same time. > > You can sterilize them with EtOH. > > > Paul Herzmark > Specialist > [hidden email] > > Department of Molecular and Cell Biology > 479 Life Science Addition > University of California, Berkeley > Berkeley, CA 94720-3200 > (510) 643-9603 > (510) 643-9500 fax > > > On Wed, Jan 7, 2009 at 6:49 PM, Melinda Larsen <[hidden email]> > wrote: >> >> I have been ordering custom-made glass-bottomed culture dishes from >> MatTek, but they have changed their manufacturing process such that it >> is fully automated and is no longer amenable to customization. Does >> anyone make their own glass-bottomed dishes by punching holes in the >> bottom of plastic culture dishes and gluing coverslips on the bottom? >> If so, could you give me some tips on how you do it? Thanks! >> >> Melinda Larsen, Ph.D., Assistant Professor >> University at Albany, SUNY >> Department of Biological Sciences > > |
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In reply to this post by Paul Herzmark
My colleague did this before. After drilling the hole, they used sand
paper to smoothen the edge.
I was reading Jim's book,and happened to find the following sentence in PC's chapter. "A rectangular hole was cut at the bottom of the Petri dish and a coverslip was glued over it with SilGaurd." (another type of glue you can use other than the one Paul mentioned) Handbook of Biological Confocal Microscopy,3rd Edition, Chapter21 Interaction of Light with Botanical Specimens, Chapter 21. P431. Hope it helps. Edna Paul Herzmark wrote: You can make gazillions easily. |
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In reply to this post by Paul Herzmark
Since I'm reading this very chapter now,Here are the possibly useful things I extracted from it.
Microspores and Pollen Grains 1. Pollen grains are usually highly absorbing, scattering and pigmented(hence strongly autofluorescenting in visible spectrum).If staining is necessary, considering NIR dyes. 2. Pollen grains/microspores can be handled the same way as suspension-cultured cells. 3. If possible, fix,stain and clear tissue with methyl-salicylate, so as to match the optical property as closely as possible to the design criteria of lens. 4. Multi-photon imaging and adaptive deconvolution might be helpful for obtaining high resolution image (Handbook of Biological Confocal Microscopy,3rd Edition, Chapter21 Interaction of Light with Botanical Specimens, Chapter 21. P431.) Edna -------------------- Hi Doug, I have been doing some 3D pollen imaging with good success. So far I imaged pollen grains up to 50 um in diameter, and it seems I get almost the same resolution on the far side as I get on the side closest to the coverslip. The sample prep was the key. 1st, the pollen I received from our pollen expert was extracted (not sure of the exact protocol, apparently somehting the pollen researchers do routinely), and stored in glacial acetic acid. The result is that the highly absorbent and scattering contents of the pollen grain has been removed, and all that is left is the pollen "shell" with its characteristic shape and features. It has enough autofluorescence for confocal imaging without any staining. The pollen is gradually re-hydrated and infiltrated with 2-2-thiodiethanol mounting medium. below is the prep protocol: The microwave just accelerates the process, you could just do longer time each step without the MW. 100 µl of the pollen suspension was placed in a microcentrifuge tube and gradually re-hydrated by adding increasing amounts of water (from 10 µl to 600 µl), pulse-mixing (vortexing) and irradiating in a Pelco Biowave (Ted Pella Inc., Redding, CA) laboratory microwave processor for 1 min at 230W to accelerate the diffusion process. Total 12 steps were performed. Pollen was then spun down by centrifugation at 500 x g for 1 min, resuspended in phosphate buffered saline (PBS; 140 mM NaCl, 3 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4) and microwaved as above. Subsequently, the pollen was gradually infiltrated with 2,2’-thiodiethanol (TDE), a mounting medium for high-resolution microscopy (Staudt et al., 2007). The pollen was spun down as above before each step. The pollen pellet was resuspended using 10%, 25%, 50% v/v TDE/PBS mixture, and finally three times in 97% v/v TDE/PBS, In each step, 1 min microwave irradiation at 230W was used after resuspension. The pollen was then spun down and the pellet resuspended and stored in 97% TDE/PBS. Confocal Microscopy (Inverted microscope) For microscopy, a droplet of the pollen suspension was applied to a coverslip- bottom imaging chamber (coverslip thickness ~175 µm) and the pollen immobilized by placing a small coverslip on the top of the droplet, with a small dab of dried nail polish on one side as a spacer so that the weight of the coverslip does not cause flattening of the pollen grain. Close to the spacer, there layer of the mounting medium is too deep, and the pollen is floating freely and cannot be imaged. On the other end of the coverslip, there is no spacer and the pollen is squished, deformed. So I try to find a position in between, where the pollen is not moving, but is not visibly squished. Imaging was done using Olympus FV1000 laser scanning confocal microscope system attached to an Olympus IX81 inverted microscope with a 100x/1.4 oil immersion objective (UPLSAPO 100x/1.4). Fluorescence was excited using 488 nm Argon ion laser, the fluorescence detector was set to 500-600nm bandpass. Laser output was programmed to increase as we go deeper from the surface in order to compensate for the loss of signal. Typically, the laser output would double from 8 to 16% for a Z-stack going from 0 to 50 um depth. Confocal scanning was performed in the photon counting mode, scan speed (pixel dwell time) was 20 µs/pixel or 10 µs/pixel. The confocal aperture was set to 120 µm, which corresponds to 0.75 Airy Unit. The confocal zoom and scan size was set to achieve 65nm pixel size in XY. The z-step was set to 130nm. Typical acquisition time was between 20 and 50 min per stack, depending on the size of the pollen grain and scan speed (i.e., signal intensity). You may not need to do such a fine XYZ step, but I have to say the resulting datasets look great; Sincerely, Stan Vitha Microscopy and Imaging Center, Texas A&M University On Tue, 6 Jan 2009 10:47:06 -0700, [hidden email] wrote: >I have very close to zero experience prepping plant tissues for any type of >microscopy. I've been contacted by a botanist that is interested in looking >at a variety of pollen types and he's never done confocal. He'd like to >look at the 3D structures and it sounds like good 3D may very well be >important. > > > >I know from limited experience that the far side of a pollen grain often >doesn't look as good in a 3D rendering, presumably due to spherical >aberration, scattering, and other optical effects. I gather from the >resources I have that pollen is highly autofluorescent across the entire VIS >spectrum (probably why the vendors love to use it for demos). > > > >Any tips, suggested review articles, web sites, etc that people could point >me to for sample prep techniques? Any microscopy caveats to be aware of? > > > >Many thanks. > >Doug > > > >^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ > >Douglas W. Cromey, M.S. - Assistant Scientific Investigator > >Dept. of Cell Biology & Anatomy, University of Arizona > >1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA > > > >office: AHSC 4212 email: [hidden email] > >voice: 520-626-2824 fax: 520-626-2097 > > > >http://swehsc.pharmacy.arizona.edu/exppath/ > >Home of: "Microscopy and Imaging Resources on the WWW" > > > > |
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vaishali kailaje |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Melinda Larsen
Hi,
You can cut the petri plate in the centre approximately 1.5-2cm. Then stick a coverslip with DPX at the bottom of the plate and allow to dry for a 4-6hrs. If details required you can write me at my mail id. Vaishali K. ACTREC, Tata Memorial Centre, India. [hidden email] --- On Thu, 1/8/09, Melinda Larsen <[hidden email]> wrote: > From: Melinda Larsen <[hidden email]> > Subject: home-made glass-bottomed cell culture dishes > To: [hidden email] > Date: Thursday, January 8, 2009, 8:19 AM > I have been ordering custom-made glass-bottomed culture > dishes from > MatTek, but they have changed their manufacturing process > such that it > is fully automated and is no longer amenable to > customization. Does > anyone make their own glass-bottomed dishes by punching > holes in the > bottom of plastic culture dishes and gluing coverslips on > the bottom? > If so, could you give me some tips on how you do it? > Thanks! > > Melinda Larsen, Ph.D., Assistant Professor > University at Albany, SUNY > Department of Biological Sciences |
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Jeremy Adler-2 |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Melinda Larsen
The easy way to make a hole in a plastic Petri dish is to heat a metal tube,
say an old cork borer, and push it through the bottom of the Petri dish. This is much easier than drilling. As an added bonus one surface of Petri dish ends up smooth and can be used to attach a cover slip, no sanding etc required. We use nail varnish to attach a cover slip which is less messy than silicon sealant. Dr Jeremy Adler F451a Cell Biologi Wenner-Gren Inst. The Arhenius Lab Stockholm University S-106 91 Stockholm Sweden tel +46 (0)8 16 2759 -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Melinda Larsen Sent: den 8 januari 2009 03:49 To: [hidden email] Subject: home-made glass-bottomed cell culture dishes I have been ordering custom-made glass-bottomed culture dishes from MatTek, but they have changed their manufacturing process such that it is fully automated and is no longer amenable to customization. Does anyone make their own glass-bottomed dishes by punching holes in the bottom of plastic culture dishes and gluing coverslips on the bottom? If so, could you give me some tips on how you do it? Thanks! Melinda Larsen, Ph.D., Assistant Professor University at Albany, SUNY Department of Biological Sciences |
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RICHARD BURRY |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Paul Herzmark
Paul
The biggest problem is parallel to the bottom of the dish! Small pieces of cut plastic will stick down from the cut edge of the hole, and when the cover slip is glued on, it will be not parallel to the bottom of the dish. The resulting dish will, when on a microscope stage, will need to be refocused as it is moved. Dick ----- Original Message ----- From: Paul Herzmark <[hidden email]> Date: Wednesday, January 7, 2009 10:50 pm Subject: Re: home-made glass-bottomed cell culture dishes To: [hidden email] > You can make gazillions easily. > Drill a big hole in the plastic dish. If you have a drill press it is assembly line speed. Circle the hole with some silicon glue (sparingly) like is used for aquariums. stick on your coverslip. Make sure it is nice a parallel to the rest of the plastic dish bottom so the whole microscope field is in focus at the same time. > You can sterilize them with EtOH. > Paul Herzmark > Specialist <A href="javascript:main.compose('new','t=pherzmark@gmail.com')">> pherzmark@... > Department of Molecular and Cell Biology > 479 Life Science Addition > University of California, Berkeley > Berkeley, CA 94720-3200 > (510) 643-9603 > (510) 643-9500 fax > On Wed, Jan 7, 2009 at 6:49 PM, Melinda Larsen <<A href="javascript:main.compose('new','t=melinda.larsen@gmail.com')">melinda.larsen@...> wrote:
> I have been ordering custom-made glass-bottomed culture dishes from |
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leoncio vergara |
Re: home-made glass-bottomed cell culture dishes
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if you use a dremel machine, with a polishing attachment, on the bottom (outside, no cell side) of the dish, you will avoid that problem. Still make sure you press the coverslip evenly around.
________________________________________ From: Confocal Microscopy List [[hidden email]] On Behalf Of RICHARD BURRY [[hidden email]] Sent: Thursday, January 08, 2009 6:34 AM To: [hidden email] Subject: Re: home-made glass-bottomed cell culture dishes Paul The biggest problem is parallel to the bottom of the dish! Small pieces of cut plastic will stick down from the cut edge of the hole, and when the cover slip is glued on, it will be not parallel to the bottom of the dish. The resulting dish will, when on a microscope stage, will need to be refocused as it is moved. Dick ----- Original Message ----- From: Paul Herzmark <[hidden email]> Date: Wednesday, January 7, 2009 10:50 pm Subject: Re: home-made glass-bottomed cell culture dishes To: [hidden email] > You can make gazillions easily. > Drill a big hole in the plastic dish. If you have a drill press it is assembly line speed. Circle the hole with some silicon glue (sparingly) like is used for aquariums. stick on your coverslip. Make sure it is nice a parallel to the rest of the plastic dish bottom so the whole microscope field is in focus at the same time. > You can sterilize them with EtOH. > Paul Herzmark > Specialist > [hidden email]<javascript:main.compose('new','t=[hidden email]')> > Department of Molecular and Cell Biology > 479 Life Science Addition > University of California, Berkeley > Berkeley, CA 94720-3200 > (510) 643-9603 > (510) 643-9500 fax > On Wed, Jan 7, 2009 at 6:49 PM, Melinda Larsen <[hidden email]<javascript:main.compose('new','t=[hidden email]')>> wrote: > I have been ordering custom-made glass-bottomed culture dishes from > MatTek, but they have changed their manufacturing process such that it > is fully automated and is no longer amenable to customization. Does > anyone make their own glass-bottomed dishes by punching holes in the > bottom of plastic culture dishes and gluing coverslips on the bottom? > If so, could you give me some tips on how you do it? Thanks! > Melinda Larsen, Ph.D., Assistant Professor > University at Albany, SUNY > Department of Biological Sciences ________________________________ |
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Gary Laevsky-2 |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Melinda Larsen
Using a Rose Chamber or a modified version thereof entails a little more up front cost but lasts forever. |
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In reply to this post by Melinda Larsen
The way we do it is described in:
D. Kline. Quantitative microinjection of mouse oocytes and eggs. pp. 139 and 144. In David Carrol (ed). Microinjection: Methods in applications. Vol 118. 2009 Humana Press -----Original Message----- From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Melinda Larsen Sent: Wednesday, January 07, 2009 9:49 PM To: [hidden email] Subject: home-made glass-bottomed cell culture dishes I have been ordering custom-made glass-bottomed culture dishes from MatTek, but they have changed their manufacturing process such that it is fully automated and is no longer amenable to customization. Does anyone make their own glass-bottomed dishes by punching holes in the bottom of plastic culture dishes and gluing coverslips on the bottom? If so, could you give me some tips on how you do it? Thanks! Melinda Larsen, Ph.D., Assistant Professor University at Albany, SUNY Department of Biological Sciences |
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Ignatius, Mike |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by RICHARD BURRY
Hi Paul,
We did something not mentioned yet, that avoided this issue
of canted or tilted coverslips. Make a donut shaped circle out of
standard lab parafilm, the size of the outer diameter of the round cover slip
and about 2-3 mm wide. We made a punch that created these, but you can
easily just cut them.
After making sure the edges of the hole in the culture
dish are smooth, you flip the dish over, place the ring/donut of
parafilm down around the outside of the hole, then lay the glass coverslip
(only glass will work here) over the parafilm/hole. Then using a heating tool,
like a standard soldering iron, lightly touch and heat the glass coverslip
edges, which readily melts the parafilm underneath. When it cools, it
forms a perfect, water tight and optically clear seal that is parallel to the
dish bottom. Fun and easy to do, with no toxic (and unevenly applied)
glues to worry about. To avoid lots of rough edges our shop milled the
holes, rather than drilling them. They will know the
difference.
Regards,
Mike Ignatius From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of RICHARD BURRY Sent: Thursday, January 08, 2009 4:34 AM To: [hidden email] Subject: Re: home-made glass-bottomed cell culture dishes The biggest problem is parallel to the bottom of the dish! Small pieces of cut plastic will stick down from the cut edge of the hole, and when the cover slip is glued on, it will be not parallel to the bottom of the dish. The resulting dish will, when on a microscope stage, will need to be refocused as it is moved. Dick ----- Original Message ----- From: Paul Herzmark <[hidden email]> Date: Wednesday, January 7, 2009 10:50 pm Subject: Re: home-made glass-bottomed cell culture dishes To: [hidden email] > You can make gazillions easily. > Drill a big hole in the plastic dish. If you have a drill press it is assembly line speed. Circle the hole with some silicon glue (sparingly) like is used for aquariums. stick on your coverslip. Make sure it is nice a parallel to the rest of the plastic dish bottom so the whole microscope field is in focus at the same time. > You can sterilize them with EtOH. > Paul Herzmark > Specialist <A href="javascript:main.compose('new','t=pherzmark@gmail.com')">> pherzmark@... > Department of Molecular and Cell Biology > 479 Life Science Addition > University of California, Berkeley > Berkeley, CA 94720-3200 > (510) 643-9603 > (510) 643-9500 fax >
On Wed, Jan 7, 2009 at 6:49 PM, Melinda Larsen <<A
href="javascript:main.compose('new','t=melinda.larsen@gmail.com')">melinda.larsen@...>
wrote:
> I have been ordering custom-made glass-bottomed culture dishes from |
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Melinda Larsen |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Knecht, David
Interesting -- I hadn't thought of using glass dishes. Thanks!
On Wed, Jan 7, 2009 at 10:53 PM, David Knecht ATT <[hidden email]> wrote: > We used to do this (now using Willco Wells). We found it easier to use > Glass Petri dishes as they could be easily cut by someone who knows how to > drill glass (our shop did this). Then the coverslips were glued onto the > bottom forming a small well in the bottom of the dish. I don't remember > what glue we used but it was not hard to reglue broken ones. Dave > On Jan 7, 2009, at 9:49 PM, Melinda Larsen wrote: > > I have been ordering custom-made glass-bottomed culture dishes from > MatTek, but they have changed their manufacturing process such that it > is fully automated and is no longer amenable to customization. Does > anyone make their own glass-bottomed dishes by punching holes in the > bottom of plastic culture dishes and gluing coverslips on the bottom? > If so, could you give me some tips on how you do it? Thanks! > > Melinda Larsen, Ph.D., Assistant Professor > University at Albany, SUNY > Department of Biological Sciences > > Dr. David Knecht > Department of Molecular and Cell Biology > Co-head Flow Cytometry and Confocal Microscopy Facility > U-3125 > 91 N. Eagleville Rd. > University of Connecticut > Storrs, CT 06269 > 860-486-2200 > 860-486-4331 (fax) > > |
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Rosemary.White |
Re: home-made glass-bottomed cell culture dishes
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Yes, glass is good because you can recycle a number of times. Haven't done
this for a while but we used to use large glass slides, about 4 cm by 8 cm by about 1.5 mm deep, and had ones with different-sized holes in them to make wells for use in different applications. You can get lots of different sizes of coverslips to make the base, and we'd seal them on with valap (1:1:1 vaseline:lanolin:paraffin), because our plant tissues don't like any nail polish near them (even the following day after overnight soaking of the nail polish-sealed wells in buffer), and so we could re-use the slides easily. We weren't growing cells on the coverslips, of course, just using them for microinjection and observation of tissues that needed to stay immersed. cheers, Rosemary Rosemary White CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia ph 61 2 6246 5475 fx 61 2 6246 5334 On 9/01/09 12:33 PM, "Melinda Larsen" <[hidden email]> wrote: > Interesting -- I hadn't thought of using glass dishes. Thanks! > > On Wed, Jan 7, 2009 at 10:53 PM, David Knecht ATT > <[hidden email]> wrote: >> We used to do this (now using Willco Wells). We found it easier to use >> Glass Petri dishes as they could be easily cut by someone who knows how to >> drill glass (our shop did this). Then the coverslips were glued onto the >> bottom forming a small well in the bottom of the dish. I don't remember >> what glue we used but it was not hard to reglue broken ones. Dave >> On Jan 7, 2009, at 9:49 PM, Melinda Larsen wrote: >> >> I have been ordering custom-made glass-bottomed culture dishes from >> MatTek, but they have changed their manufacturing process such that it >> is fully automated and is no longer amenable to customization. Does >> anyone make their own glass-bottomed dishes by punching holes in the >> bottom of plastic culture dishes and gluing coverslips on the bottom? >> If so, could you give me some tips on how you do it? Thanks! >> >> Melinda Larsen, Ph.D., Assistant Professor >> University at Albany, SUNY >> Department of Biological Sciences >> >> Dr. David Knecht >> Department of Molecular and Cell Biology >> Co-head Flow Cytometry and Confocal Microscopy Facility >> U-3125 >> 91 N. Eagleville Rd. >> University of Connecticut >> Storrs, CT 06269 >> 860-486-2200 >> 860-486-4331 (fax) >> >> |
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Farid Jalali |
Re: home-made glass-bottomed cell culture dishes
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I'm not sure what a Rose Chamber is but the Atto-chamber that
Molecular Probes/ Invitrogen sells is a fantastic investment. I switched to this from coverslip bottom dishes and found it very easy to use and it will last forever. No commercial interest. Best Farid On 1/8/09, Rosemary White <[hidden email]> wrote: > Yes, glass is good because you can recycle a number of times. Haven't done > this for a while but we used to use large glass slides, about 4 cm by 8 cm > by about 1.5 mm deep, and had ones with different-sized holes in them to > make wells for use in different applications. You can get lots of different > sizes of coverslips to make the base, and we'd seal them on with valap > (1:1:1 vaseline:lanolin:paraffin), because our plant tissues don't like any > nail polish near them (even the following day after overnight soaking of the > nail polish-sealed wells in buffer), and so we could re-use the slides > easily. We weren't growing cells on the coverslips, of course, just using > them for microinjection and observation of tissues that needed to stay > immersed. > cheers, > Rosemary > > > Rosemary White > CSIRO Plant Industry > GPO Box 1600 > Canberra, ACT 2601 > Australia > > ph 61 2 6246 5475 > fx 61 2 6246 5334 > > On 9/01/09 12:33 PM, "Melinda Larsen" <[hidden email]> wrote: > >> Interesting -- I hadn't thought of using glass dishes. Thanks! >> >> On Wed, Jan 7, 2009 at 10:53 PM, David Knecht ATT >> <[hidden email]> wrote: >>> We used to do this (now using Willco Wells). We found it easier to use >>> Glass Petri dishes as they could be easily cut by someone who knows how >>> to >>> drill glass (our shop did this). Then the coverslips were glued onto the >>> bottom forming a small well in the bottom of the dish. I don't remember >>> what glue we used but it was not hard to reglue broken ones. Dave >>> On Jan 7, 2009, at 9:49 PM, Melinda Larsen wrote: >>> >>> I have been ordering custom-made glass-bottomed culture dishes from >>> MatTek, but they have changed their manufacturing process such that it >>> is fully automated and is no longer amenable to customization. Does >>> anyone make their own glass-bottomed dishes by punching holes in the >>> bottom of plastic culture dishes and gluing coverslips on the bottom? >>> If so, could you give me some tips on how you do it? Thanks! >>> >>> Melinda Larsen, Ph.D., Assistant Professor >>> University at Albany, SUNY >>> Department of Biological Sciences >>> >>> Dr. David Knecht >>> Department of Molecular and Cell Biology >>> Co-head Flow Cytometry and Confocal Microscopy Facility >>> U-3125 >>> 91 N. Eagleville Rd. >>> University of Connecticut >>> Storrs, CT 06269 >>> 860-486-2200 >>> 860-486-4331 (fax) >>> >>> > -- Sent from my mobile device |
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Chris Wood-5 |
Re: home-made glass-bottomed cell culture dishes
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In reply to this post by Melinda Larsen
Cheaper alternatives to the Attofluor chamber are made by Harvard
instruments (cat # 640370), but not autoclavable. We tried sterilising them with alchohol and UV exposure but this never reliably worked - we'd end up losing too many experiments with contamination (won't be too problemantic for short experiments, but we routinely image cells for 24hrs+) for it to be worth the cost savings. We also tried the drilling and sticking 100% homemade approach but our ES cells didn't grow the same, we suspected some toxicity from the glue (we tested a few). We now use the Fluorodish from WPI which has a large working area available on the coverslip (23mm) and the ES cells seem happy to grow there. The boss is starting to grumble about the cost though, so I might try the Attofluor approach with an all metal (autoclavable) chamber, but 270 USD for each one isn't very friendly, four or five would be the minimum required for us. I think I'll just buy the nitrile o-rings and get a local machine shop to copy the design instead. I like the parafilm-soldering iron idea as well, will definitely give it a spin. Saludos Chris Instituto de Biotecnología Universidad Nacional Autónoma de Mexico |
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