suggestions for cutting through plastic

[Jerry (Gerald) Sedgewick]

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Hello all,

I'm doing a demonstration for live cell imaging using an inverted,
fluorescence microscope.  I'd like to do a fairly low tech method for
the first demo.  My thought is to use a plastic culture dish, cut a
square in the bottom, and then glue a cover slip to the bottom.  I will
be filling it with pond water since I should find plenty of
autofluorescent organisms.

I recall having tried to cut through a plastic culture dish and it
wasn't easy.  I'm wondering if any of you have ever tried this and have
had success at cutting through this sort of dish.  How did you do it
while retaining smooth edges?

Any other means for accomplishing the same end would be appreciated for
a project without a budget.

Thanks!

Jerry Sedgewick

[Paul Herzmark]

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Drill a hole and glue on your cover slip. Easy!

Paul Herzmark


On Tue, Jun 4, 2013 at 7:10 PM, jerry sedgewick <[hidden email]>wrote:

> *****
> 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>
> *****
>
> Hello all,
>
> I'm doing a demonstration for live cell imaging using an inverted,
> fluorescence microscope.  I'd like to do a fairly low tech method for the
> first demo.  My thought is to use a plastic culture dish, cut a square in
> the bottom, and then glue a cover slip to the bottom.  I will be filling it
> with pond water since I should find plenty of autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it wasn't
> easy.  I'm wondering if any of you have ever tried this and have had
> success at cutting through this sort of dish.  How did you do it while
> retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated for a
> project without a budget.
>
> Thanks!
>
> Jerry Sedgewick
>

[Matthew Nicholas-2]

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Hi Jerry,


On 6/5/2013 12:45 AM, Paul Herzmark wrote:
>
> Drill a hole and glue on your cover slip. Easy!
>
> Paul Herzmark
I agree with Paul's suggestion. Tip: to avoid cracking the dish, use a
sharp bit and a wooden backing. Hold or clamp the dish firmly to the
wood. Drill through the dish and into the wood.

>
>
> On Tue, Jun 4, 2013 at 7:10 PM, jerry sedgewick <[hidden email]>wrote:
>
>> I recall having tried to cut through a plastic culture dish and it wasn't
>> easy.  I'm wondering if any of you have ever tried this and have had
>> success at cutting through this sort of dish.  How did you do it while
>> retaining smooth edges?

Other things I've tried in the past to cut plastic with decent success
(though not specifically culture dishes, and usually with worse results
than drilling):

a) dremel tool with a rotary cutting blade (use a relatively slow speed
to avoid melting the plastic as you cut. The edge can be smoothed
somewhat (if needed) by very carefully and sparingly running a flame
over it (obviously you should work in a fume hood, away from any
flammable materials/solvents, and be extremely careful). Alternatively,
you can sand the edges with a very fine-grit sandpaper, and then clean
the dish well to remove any dust.

b) a hot blade. In the past I've used an xacto blade attached to a
soldering iron (I believe you can buy these; otherwise you'll have to
cobble something together to attach the blade). This works fairly well,
as long as the iron is fairly powerful and can transfer quite a bit of
heat to the blade. In a pinch I have used a hot wire, but with worse
results.

c) if you have an appropriate metal shape (e.g. a square pipe) of the
size and shape you want to cut out, you can carefully heat the end (hold
it with a pliers) to very high temperature and then press it through the
plastic.

[Jeremy Adler-4]

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Heat the end of a metal tube in a Bunsen burnfer and then getnly push  
the end thru the base of the petri dish and then pull it out. The hole  
on one side has a clean smooth edge - stick on the glass coverslip.  
Cutters used for making holes in rubber bungs are effective.



Quoting jerry sedgewick <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello all,
>
> I'm doing a demonstration for live cell imaging using an inverted,  
> fluorescence microscope.  I'd like to do a fairly low tech method  
> for the first demo.  My thought is to use a plastic culture dish,  
> cut a square in the bottom, and then glue a cover slip to the  
> bottom.  I will be filling it with pond water since I should find  
> plenty of autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it  
> wasn't easy.  I'm wondering if any of you have ever tried this and  
> have had success at cutting through this sort of dish.  How did you  
> do it while retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated  
> for a project without a budget.
>
> Thanks!
>
> Jerry Sedgewick
>

Jeremy Adler
IGP
Rudbeckslaboratoriet
Daghammersköljdsväg 20
751 85 Uppsala
Sweden

0046 (0)18 471 4607

[Mark Cannell-2]

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Sharp drill and finish with a Dremel barrel sanding bit.

HTH

Mark

On 5/06/2013, at 8:48 AM, Jeremy Adler <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Heat the end of a metal tube in a Bunsen burnfer and then getnly push the end thru the base of the petri dish and then pull it out. The hole on one side has a clean smooth edge - stick on the glass coverslip. Cutters used for making holes in rubber bungs are effective.
>
>
>
> Quoting jerry sedgewick <[hidden email]>:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hello all,
>>
>> I'm doing a demonstration for live cell imaging using an inverted, fluorescence microscope.  I'd like to do a fairly low tech method for the first demo.  My thought is to use a plastic culture dish, cut a square in the bottom, and then glue a cover slip to the bottom.  I will be filling it with pond water since I should find plenty of autofluorescent organisms.
>>
>> I recall having tried to cut through a plastic culture dish and it wasn't easy.  I'm wondering if any of you have ever tried this and have had success at cutting through this sort of dish.  How did you do it while retaining smooth edges?
>>
>> Any other means for accomplishing the same end would be appreciated for a project without a budget.
>>
>> Thanks!
>>
>> Jerry Sedgewick
>>
>
>
>
> Jeremy Adler
> IGP
> Rudbeckslaboratoriet
> Daghammersköljdsväg 20
> 751 85 Uppsala
> Sweden
>
> 0046 (0)18 471 4607

Mark  B. Cannell Ph.D. FRSNZ
Professor of Cardiac Cell Biology
School of Physiology &  Pharmacology
Medical Sciences Building
University of Bristol
Bristol
BS8 1TD UK

[hidden email]

[Martin Wessendorf-2]

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Hey Jerry--

On 6/4/2013 9:10 PM, jerry sedgewick wrote:

> I'm doing a demonstration for live cell imaging using an inverted,
> fluorescence microscope. I'd like to do a fairly low tech method for the
> first demo. My thought is to use a plastic culture dish, cut a square in
> the bottom, and then glue a cover slip to the bottom. I will be filling
> it with pond water since I should find plenty of autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it
> wasn't easy. I'm wondering if any of you have ever tried this and have
> had success at cutting through this sort of dish. How did you do it
> while retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated for
> a project without a budget.

Even lower-tech solution: super-glue (or Scotch-tape) a longer (24 x 50
maybe) cover slip cross-wise to a microscope slide, so that one end
sticks way out from the middle of the slide like a diving board.  Then
just put a drop or two of pond-water on it.  Keep the water from seeping
out over the edges by drawing a circle of petroleum jelly on the cover
slip beforehand with a cotton applicator.  Stop evaporation, if that's
important, by sticking a second (small) cover slip on top.  --Might not
work so well with an oil condenser but it should be fine for a dry one,
as long as your mechanical stage has sufficient front-to-back travel.
May also work as a pretty good seismograph, so minimizing vibration will
be important.

Good luck!

Martin
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Dan Focht]

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Jerry


Here is another approach.
I have done this many times.
Make a retainer to support a coverslip of what ever size you want to use
or have available.
This you only have to do once because you will re-use it.
Place your coverslip in your retainer then place one of our glass
Culture Cylinders in the middle of the coverslip then add your specimen.
The botom surface of the Culture Cylinder is optically flat and
pollished so that it forms a hydrostatic seal onto the coverslip without
needing grease/wax or any other adhesive.
Link:
http://www.bioptechs.com/Products/Delta_T/Options/options.html#Culture%20Cylinders
If you want to try one, email me I'll send you a sample.
They are available with an ID from 1 to 20mm.


Dan



On 6/4/2013 10:10 PM, jerry sedgewick wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello all,
>
> I'm doing a demonstration for live cell imaging using an inverted,
> fluorescence microscope.  I'd like to do a fairly low tech method for
> the first demo.  My thought is to use a plastic culture dish, cut a
> square in the bottom, and then glue a cover slip to the bottom.  I
> will be filling it with pond water since I should find plenty of
> autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it
> wasn't easy.  I'm wondering if any of you have ever tried this and
> have had success at cutting through this sort of dish.  How did you do
> it while retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated
> for a project without a budget.
>
> Thanks!
>
> Jerry Sedgewick
>
>

--
Dan Focht Bioptechs Inc. V: [phone] www.bioptechs.com

[Travis, Jeffrey L]

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All of the previous suggestions are good and will work, and many have the advantage of requiring the use of power tools.  However, there is a simpler way:  I would suggest that you simply put a drop of your pond water in a slide and cover it with a coverslip on which you have placed small (0.05 - 0.1 mm) plasticene (modeling clay) feet to hold the slip up.  You can easily then wick in enough fluid to fill the  "chamber."  Seal the edges with molten valap or other sealant and you have a preparation that can last for days.

Regards, Jeff Travis

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of jerry sedgewick
Sent: Tuesday, June 04, 2013 10:10 PM
To: [hidden email]
Subject: suggestions for cutting through plastic

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Hello all,

I'm doing a demonstration for live cell imaging using an inverted, fluorescence microscope.  I'd like to do a fairly low tech method for the first demo.  My thought is to use a plastic culture dish, cut a square in the bottom, and then glue a cover slip to the bottom.  I will be filling it with pond water since I should find plenty of autofluorescent organisms.

I recall having tried to cut through a plastic culture dish and it wasn't easy.  I'm wondering if any of you have ever tried this and have had success at cutting through this sort of dish.  How did you do it while retaining smooth edges?

Any other means for accomplishing the same end would be appreciated for a project without a budget.

Thanks!

Jerry Sedgewick

[Eric Shelden]

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We do it this way in my lab: To drill a clean hole in the bottom of a plastic
dish, figure out some way to hold it safely on a drill press (or hold it very, very
careful). I made a holder by drilling a hole the size of the dish near one end of
a long(ish) piece of wood, and then cutting the piece of wood lengthwise
down the middle as far as one side of the hole. Then, you can hold the dish by
using a clamp to tighten across the piece of wood (making the slit you've cut
smaller) and the whole thing gets clamped to the drill press in the right
position. But for a small number of dishes, I sometimes just hold the dish.
Watch your fingers. Chuck a sharp router bit of the appropriate diameter in
the drill press and use it to create the hole by slowly lowering the bit into the
dish. Alternatively, if you have access to a metal lath, you can drill holes using
that. The point is to scrape the plastic away in a hole shape. Standard drill
bits dont work well in my experience. We use Dow corning sylgard (comes as
two liquids that you mix together) and dab some around the hole. Then put a
coverslip over  the hole and put in an oven at 50-60 degrees C until the
sylgard hardens.

[Mike Ignatius-2]

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An easy way to secure a glass (only works with glass) cover slip to the
bottom of a tissue culture (cine) dish is by cutting a parafilm 'donut',
placing it between the dish and coverslip, then sealing it with heat from a
standard soldering gun.

The parafilm makes a water tight, optically clear, long lasting seal.  You
know it is done when the parafilm turns clear from the heat - takes less
than 30 seconds.  

Mike Ignatius
Marker Gene Tech

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Eric Shelden
Sent: Wednesday, June 05, 2013 8:15 AM
To: [hidden email]
Subject: Re: suggestions for cutting through plastic

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We do it this way in my lab: To drill a clean hole in the bottom of a
plastic dish, figure out some way to hold it safely on a drill press (or
hold it very, very careful). I made a holder by drilling a hole the size of
the dish near one end of a long(ish) piece of wood, and then cutting the
piece of wood lengthwise down the middle as far as one side of the hole.
Then, you can hold the dish by using a clamp to tighten across the piece of
wood (making the slit you've cut
smaller) and the whole thing gets clamped to the drill press in the right
position. But for a small number of dishes, I sometimes just hold the dish.
Watch your fingers. Chuck a sharp router bit of the appropriate diameter in
the drill press and use it to create the hole by slowly lowering the bit
into the dish. Alternatively, if you have access to a metal lath, you can
drill holes using that. The point is to scrape the plastic away in a hole
shape. Standard drill bits dont work well in my experience. We use Dow
corning sylgard (comes as two liquids that you mix together) and dab some
around the hole. Then put a coverslip over  the hole and put in an oven at
50-60 degrees C until the sylgard hardens.

[Julio Vazquez]

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Just a few more ideas:

We use a metal slide with a hole in it, drilled by a machine shop (or a friend with a drill... I have one who does copper siding and has all that's needed...). we just smear a bit of oil around the hole underneath the slide and apply the coverslip. Coverslip is firmly sealed, and creates a small well that has enough volume for many applications (I imaged live Drosophila spermatocytes for up to 12 hours using this). Good thing is the metal slide will last forever... you only need coverslips and oil. You may be able to find a suitable pre-made piece of metal at your local hardware store that will do the job, as long as it's flat, can fit on your stage, and has a hole of a suitable size in it.

Alternatively, Wold Precision Instruments sells nice coverslip bottom dishes (cost about $ 1 / dish)... obviously, you'll need to buy a whole box, but you'll have a supply for a long time; we use a lot of these:

http://www.wpiinc.com/index.php/FD35-100.html 

Alternatively, you may be able to get free samples of microscopy vessels from the major vendors...


Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109

http://www.fhcrc.org/en.html





On Jun 4, 2013, at 7:10 PM, jerry sedgewick wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello all,
>
> I'm doing a demonstration for live cell imaging using an inverted, fluorescence microscope.  I'd like to do a fairly low tech method for the first demo.  My thought is to use a plastic culture dish, cut a square in the bottom, and then glue a cover slip to the bottom.  I will be filling it with pond water since I should find plenty of autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it wasn't easy.  I'm wondering if any of you have ever tried this and have had success at cutting through this sort of dish.  How did you do it while retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated for a project without a budget.
>
> Thanks!
>
> Jerry Sedgewick

[Craig Brideau]

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Ibidi or Mattek are often willing to send free samples, so if you only need
one or two this might be easiest.  The first one is free... @:-)

Craig
 On 2013-06-05 10:24 AM, "Mike Ignatius" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> An easy way to secure a glass (only works with glass) cover slip to the
> bottom of a tissue culture (cine) dish is by cutting a parafilm 'donut',
> placing it between the dish and coverslip, then sealing it with heat from a
> standard soldering gun.
>
> The parafilm makes a water tight, optically clear, long lasting seal.  You
> know it is done when the parafilm turns clear from the heat - takes less
> than 30 seconds.
>
> Mike Ignatius
> Marker Gene Tech
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On
> Behalf Of Eric Shelden
> Sent: Wednesday, June 05, 2013 8:15 AM
> To: [hidden email]
> Subject: Re: suggestions for cutting through plastic
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> We do it this way in my lab: To drill a clean hole in the bottom of a
> plastic dish, figure out some way to hold it safely on a drill press (or
> hold it very, very careful). I made a holder by drilling a hole the size of
> the dish near one end of a long(ish) piece of wood, and then cutting the
> piece of wood lengthwise down the middle as far as one side of the hole.
> Then, you can hold the dish by using a clamp to tighten across the piece of
> wood (making the slit you've cut
> smaller) and the whole thing gets clamped to the drill press in the right
> position. But for a small number of dishes, I sometimes just hold the dish.
> Watch your fingers. Chuck a sharp router bit of the appropriate diameter in
> the drill press and use it to create the hole by slowly lowering the bit
> into the dish. Alternatively, if you have access to a metal lath, you can
> drill holes using that. The point is to scrape the plastic away in a hole
> shape. Standard drill bits dont work well in my experience. We use Dow
> corning sylgard (comes as two liquids that you mix together) and dab some
> around the hole. Then put a coverslip over  the hole and put in an oven at
> 50-60 degrees C until the sylgard hardens.
>

[Sathya]

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Hi,
One of our facility users uses small plastic petri dishes with holes covered with cover slips for microscopy imaging and they make their own dishes. They use Vari-bit step drill bits which makes a clean hole (LENOX VARI-BIT STEP DRILL BITS- http://www.lenoxtools.com/Pages/Product.aspx?productId=VaribitStepDrillBits#ltImg[mixed]/1/). You can make smallest to large hole covering the whole dish depending on your requirement. They even create multiple small holes in the petri dish to have different samples separated from one another for different conditions within the same dish. They use the clear silicone waterproof sealant to glue the cover slips to the bottom. These sealants are non-toxic and safe to living organisms. The silicone sealant can be purchased from any aquarium or pet shop. Good luck!
 
Sathya Srinivasan
Manager
RUN Microscopy Facility
(www.ucalgary.ca/runcore)
University of Calgary
Calgary, Alberta T3L3C5
Canada
 

> Date: Tue, 4 Jun 2013 21:10:16 -0500
> From: [hidden email]
> Subject: suggestions for cutting through plastic
> To: [hidden email]
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello all,
>
> I'm doing a demonstration for live cell imaging using an inverted,
> fluorescence microscope. I'd like to do a fairly low tech method for
> the first demo. My thought is to use a plastic culture dish, cut a
> square in the bottom, and then glue a cover slip to the bottom. I will
> be filling it with pond water since I should find plenty of
> autofluorescent organisms.
>
> I recall having tried to cut through a plastic culture dish and it
> wasn't easy. I'm wondering if any of you have ever tried this and have
> had success at cutting through this sort of dish. How did you do it
> while retaining smooth edges?
>
> Any other means for accomplishing the same end would be appreciated for
> a project without a budget.
>
> Thanks!
>
> Jerry Sedgewick    

[Jerry (Gerald) Sedgewick]

*****
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Thanks everyone for your great suggestions!  Little did I know that so
many have done so much with so little!

Jerry


On 6/5/2013 4:19 PM, Sathya Srinivasan wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi,
> One of our facility users uses small plastic petri dishes with holes covered with cover slips for microscopy imaging and they make their own dishes. They use Vari-bit step drill bits which makes a clean hole (LENOX VARI-BIT STEP DRILL BITS- http://www.lenoxtools.com/Pages/Product.aspx?productId=VaribitStepDrillBits#ltImg[mixed]/1/). You can make smallest to large hole covering the whole dish depending on your requirement. They even create multiple small holes in the petri dish to have different samples separated from one another for different conditions within the same dish. They use the clear silicone waterproof sealant to glue the cover slips to the bottom. These sealants are non-toxic and safe to living organisms. The silicone sealant can be purchased from any aquarium or pet shop. Good luck!
>  
> Sathya Srinivasan
> Manager
> RUN Microscopy Facility
> (www.ucalgary.ca/runcore)
> University of Calgary
> Calgary, Alberta T3L3C5
> Canada
>  
>> Date: Tue, 4 Jun 2013 21:10:16 -0500
>> From: [hidden email]
>> Subject: suggestions for cutting through plastic
>> To: [hidden email]
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hello all,
>>
>> I'm doing a demonstration for live cell imaging using an inverted,
>> fluorescence microscope. I'd like to do a fairly low tech method for
>> the first demo. My thought is to use a plastic culture dish, cut a
>> square in the bottom, and then glue a cover slip to the bottom. I will
>> be filling it with pond water since I should find plenty of
>> autofluorescent organisms.
>>
>> I recall having tried to cut through a plastic culture dish and it
>> wasn't easy. I'm wondering if any of you have ever tried this and have
>> had success at cutting through this sort of dish. How did you do it
>> while retaining smooth edges?
>>
>> Any other means for accomplishing the same end would be appreciated for
>> a project without a budget.
>>
>> Thanks!
>>
>> Jerry Sedgewick