Simple animal demo specimen ideas

[John Runions]

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Dear microscopists,

Each year at our university Science Bazaar we demonstrate our imaging
'prowess' and equipment to the community at large (adults and kids).  This
event is always well received and the biggest attraction of all is usually
the dissecting microscopes because we let the kids look at their disgusting
fingernails and they like that.  SEMs work well because we can show head
lice and rats tongue and... well you are probably starting to see a theme.

The problem is giving a really good demo with the confocal microscopes - we
show GFP labelled organelles moving around inside living cells and demos of
3D reconstruction but this doesn't really seem to capture the kids
imaginations.

Does anybody have a favorite organism or staining technique that is simple
and effective as a demo in a situation where lots of people are moving
through fairly quickly.

I appreciate your suggestions.

John

--
John Runions, PhD
Senior Lecturer in Cell Biology
Oxford Brookes University
School of Life Sciences
Gipsy Lane, Oxford
OX3 0BP
UK

01865 483 964
http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

[kspencer007]

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I've used pollen for just this type of demo. A variety of your local flora makes it more real, and the confocal is absolutely necessary to reveal the structure, instead of being just a bright blob. Autofluorescent = no prep time. I just took some sticky tape out to some plants around the building, put mounting media on it, and covered with a coverslip. If you can show a Widefield, unresolved blob side-by-side, it is effective. Experiment beforehand to find the most interesting and diverse grains in your area, and bring in a cutting from that plant.
        Kathy
The Scripps Research Institute
La Jolla, CA


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Runions
Sent: Friday, January 28, 2011 9:11 AM
To: [hidden email]
Subject: Simple animal demo specimen ideas

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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Dear microscopists,

Each year at our university Science Bazaar we demonstrate our imaging
'prowess' and equipment to the community at large (adults and kids).  This
event is always well received and the biggest attraction of all is usually
the dissecting microscopes because we let the kids look at their disgusting
fingernails and they like that.  SEMs work well because we can show head
lice and rats tongue and... well you are probably starting to see a theme.

The problem is giving a really good demo with the confocal microscopes - we
show GFP labelled organelles moving around inside living cells and demos of
3D reconstruction but this doesn't really seem to capture the kids
imaginations.

Does anybody have a favorite organism or staining technique that is simple
and effective as a demo in a situation where lots of people are moving
through fairly quickly.

I appreciate your suggestions.

John

--
John Runions, PhD
Senior Lecturer in Cell Biology
Oxford Brookes University
School of Life Sciences
Gipsy Lane, Oxford
OX3 0BP
UK

01865 483 964
http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

[Boswell, Carl A - (cboswell)]

Primary culture of neonatal rat cardiomyocytes (or chick embryos, etc.) will beat spontaneously, either individually, or as sheets if dense enough.  GFP labeling of the right structural protein will illustrate the striations in the live cells.  Very bizarre to see under the microscope.  
C


Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
Univ. of Arizona
520-954-7053
FAX [phone]


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Runions
Sent: Friday, January 28, 2011 10:11 AM
To: [hidden email]
Subject: Simple animal demo specimen ideas

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear microscopists,

Each year at our university Science Bazaar we demonstrate our imaging 'prowess' and equipment to the community at large (adults and kids).  This event is always well received and the biggest attraction of all is usually the dissecting microscopes because we let the kids look at their disgusting fingernails and they like that.  SEMs work well because we can show head lice and rats tongue and... well you are probably starting to see a theme.

The problem is giving a really good demo with the confocal microscopes - we show GFP labelled organelles moving around inside living cells and demos of 3D reconstruction but this doesn't really seem to capture the kids imaginations.

Does anybody have a favorite organism or staining technique that is simple and effective as a demo in a situation where lots of people are moving through fairly quickly.

I appreciate your suggestions.

John

--
John Runions, PhD
Senior Lecturer in Cell Biology
Oxford Brookes University
School of Life Sciences
Gipsy Lane, Oxford
OX3 0BP
UK

01865 483 964
http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

[Knecht, David]

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I always aim for cells doing something comprehensible as a process in these demos.  I use Dictyostelium because it can be done at ambient temperature and there are so many behaviors to look at.
1.  Dictyostelium cells crawling randomly or moving in a chemotactic gradient, labeled with a GFP-actin binding protein allows you to see protrusions form as the cells move.  
2.  The cells in #1 will also macropinocytose constitutively, and the actin probe labels the forming macropinosome transiently.  If you put fluorscent dextran or quantum dots in the medium, you can watch uptake, processing and movement of macropinosomes.
3.  If you add yeast, you can watch the cells phagocytose the yeast.  That engulfment/eating is one kids particularly like because they can relate to eating.
4.  Streaming aggregation with GFP labeled cells mixed with unlabeled cells is an easy way to show multicellular behavior.  The pulsatile nature of the movement, the spiral galaxy arms of the aggregation process and the crawling slugs all seem to stimulate interest.  
You can see examples of some of these on my class website: http://homepages.uconn.edu/~mb2225vc/MCB_2225/

On Jan 28, 2011, at 12:10 PM, John Runions wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear microscopists,
>
> Each year at our university Science Bazaar we demonstrate our imaging
> 'prowess' and equipment to the community at large (adults and kids).  This
> event is always well received and the biggest attraction of all is usually
> the dissecting microscopes because we let the kids look at their disgusting
> fingernails and they like that.  SEMs work well because we can show head
> lice and rats tongue and... well you are probably starting to see a theme.
>
> The problem is giving a really good demo with the confocal microscopes - we
> show GFP labelled organelles moving around inside living cells and demos of
> 3D reconstruction but this doesn't really seem to capture the kids
> imaginations.
>
> Does anybody have a favorite organism or staining technique that is simple
> and effective as a demo in a situation where lots of people are moving
> through fairly quickly.
>
> I appreciate your suggestions.
>
> John
>
> --
> John Runions, PhD
> Senior Lecturer in Cell Biology
> Oxford Brookes University
> School of Life Sciences
> Gipsy Lane, Oxford
> OX3 0BP
> UK
>
> 01865 483 964
> http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

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)

[Glen MacDonald-2]

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Cardiomyocytes will also load with Fluo-4 AM or Fluo-3 AM, in addition to the usual organelle suspects from the Molecular Probes catalog, such as Mitotrackers.
An anesthetized zebrafish larva or xenopus tadpole in a blob of 1-2% agarose will show blood flow, heart, then try reflection or autofluorescence.  You might find transgenics.  I can give you a list of labels specific for neuromast sensory organs, inner ear and nasal epithelium.  

Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923  USA
[phone]
[hidden email]








On Jan 28, 2011, at 9:29 AM, Boswell, Carl A - (cboswell) wrote:

> Primary culture of neonatal rat cardiomyocytes (or chick embryos, etc.) will beat spontaneously, either individually, or as sheets if dense enough.  GFP labeling of the right structural protein will illustrate the striations in the live cells.  Very bizarre to see under the microscope.  
> C
>
>
> Carl A. Boswell, Ph.D.
> Molecular and Cellular Biology
> Univ. of Arizona
> 520-954-7053
> FAX [phone]
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Runions
> Sent: Friday, January 28, 2011 10:11 AM
> To: [hidden email]
> Subject: Simple animal demo specimen ideas
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear microscopists,
>
> Each year at our university Science Bazaar we demonstrate our imaging 'prowess' and equipment to the community at large (adults and kids).  This event is always well received and the biggest attraction of all is usually the dissecting microscopes because we let the kids look at their disgusting fingernails and they like that.  SEMs work well because we can show head lice and rats tongue and... well you are probably starting to see a theme.
>
> The problem is giving a really good demo with the confocal microscopes - we show GFP labelled organelles moving around inside living cells and demos of 3D reconstruction but this doesn't really seem to capture the kids imaginations.
>
> Does anybody have a favorite organism or staining technique that is simple and effective as a demo in a situation where lots of people are moving through fairly quickly.
>
> I appreciate your suggestions.
>
> John
>
> --
> John Runions, PhD
> Senior Lecturer in Cell Biology
> Oxford Brookes University
> School of Life Sciences
> Gipsy Lane, Oxford
> OX3 0BP
> UK
>
> 01865 483 964
> http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!
>

[Casey Laris]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I've done this kind of demo with zerbrafish. We did it with transfected live
animals expressing GFP in the heart. It was fun watching the beating heart
under video rate fluorescence at low power - maybe 4X - 10X. UV excitation
should work for autofluorescence too. Confocal isn't exactly required but
this is a definite crowd pleaser if you have access to something similar.

I agree that cardiomyocytes with Fluo-4 would be a good idea too. A process
connected to a beating heart seems an accessible concept for the local
community at large. Here is a link to a video showing transients in action.

http://youtu.be/4Gi7HS3rmwc?hd=1  <http://youtu.be/4Gi7HS3rmwc?hd=1>

I apologize that the tone of the video is on the commercial side for this
forum - and I do have personal interest in the company - but the first 45
seconds show nice examples of calcium transients from cardiomyocytes. The
preps aren't trivial here either but it's something a heart lab should be
able to pull together for you.

Best,

Casey Laris | Vala Sciences
858.461.6862 | www.valasciences.com
[hidden email]


On Fri, Jan 28, 2011 at 10:06 AM, Glen MacDonald
<[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Cardiomyocytes will also load with Fluo-4 AM or Fluo-3 AM, in addition to
> the usual organelle suspects from the Molecular Probes catalog, such as
> Mitotrackers.
> An anesthetized zebrafish larva or xenopus tadpole in a blob of 1-2%
> agarose will show blood flow, heart, then try reflection or
> autofluorescence.  You might find transgenics.  I can give you a list of
> labels specific for neuromast sensory organs, inner ear and nasal
> epithelium.
>
> Regards,
> Glen
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923  USA
> [phone]
> [hidden email]
>
>
>
>
>
>
>
>
> On Jan 28, 2011, at 9:29 AM, Boswell, Carl A - (cboswell) wrote:
>
> > Primary culture of neonatal rat cardiomyocytes (or chick embryos, etc.)
> will beat spontaneously, either individually, or as sheets if dense enough.
>  GFP labeling of the right structural protein will illustrate the striations
> in the live cells.  Very bizarre to see under the microscope.
> > C
> >
> >
> > Carl A. Boswell, Ph.D.
> > Molecular and Cellular Biology
> > Univ. of Arizona
> > 520-954-7053
> > FAX [phone]
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of John Runions
> > Sent: Friday, January 28, 2011 10:11 AM
> > To: [hidden email]
> > Subject: Simple animal demo specimen ideas
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Dear microscopists,
> >
> > Each year at our university Science Bazaar we demonstrate our imaging
> 'prowess' and equipment to the community at large (adults and kids).  This
> event is always well received and the biggest attraction of all is usually
> the dissecting microscopes because we let the kids look at their disgusting
> fingernails and they like that.  SEMs work well because we can show head
> lice and rats tongue and... well you are probably starting to see a theme.
> >
> > The problem is giving a really good demo with the confocal microscopes -
> we show GFP labelled organelles moving around inside living cells and demos
> of 3D reconstruction but this doesn't really seem to capture the kids
> imaginations.
> >
> > Does anybody have a favorite organism or staining technique that is
> simple and effective as a demo in a situation where lots of people are
> moving through fairly quickly.
> >
> > I appreciate your suggestions.
> >
> > John
> >
> > --
> > John Runions, PhD
> > Senior Lecturer in Cell Biology
> > Oxford Brookes University
> > School of Life Sciences
> > Gipsy Lane, Oxford
> > OX3 0BP
> > UK
> >
> > 01865 483 964
> > http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!
> >
>

[Ignatius, Mike-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

If you can a prepared slide of an insect pupa, the autofluorescence allows fun 3D viewing.  With wide field you just see the outside of the pupa, but with Confocal sectioning the insect inside appears.   Then you can reconstruct the animal with max projection are really sense the whole body structure.  And no dyes or bleaching or such to be concerned with.  

This site has 200 slides on sale for $89 with a few examples of larval forms but many more that could be fun to play with.  http://store.amscope.com/ps200.html   Can't beat the price.

Mike Ignatius
Molecular Probes, Life Technologies





-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Casey Laris
Sent: Friday, January 28, 2011 11:39 AM
To: [hidden email]
Subject: Re: Simple animal demo specimen ideas

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I've done this kind of demo with zerbrafish. We did it with transfected live
animals expressing GFP in the heart. It was fun watching the beating heart
under video rate fluorescence at low power - maybe 4X - 10X. UV excitation
should work for autofluorescence too. Confocal isn't exactly required but
this is a definite crowd pleaser if you have access to something similar.

I agree that cardiomyocytes with Fluo-4 would be a good idea too. A process
connected to a beating heart seems an accessible concept for the local
community at large. Here is a link to a video showing transients in action.

http://youtu.be/4Gi7HS3rmwc?hd=1  <http://youtu.be/4Gi7HS3rmwc?hd=1>

I apologize that the tone of the video is on the commercial side for this
forum - and I do have personal interest in the company - but the first 45
seconds show nice examples of calcium transients from cardiomyocytes. The
preps aren't trivial here either but it's something a heart lab should be
able to pull together for you.

Best,

Casey Laris | Vala Sciences
858.461.6862 | www.valasciences.com
[hidden email]


On Fri, Jan 28, 2011 at 10:06 AM, Glen MacDonald
<[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Cardiomyocytes will also load with Fluo-4 AM or Fluo-3 AM, in addition to
> the usual organelle suspects from the Molecular Probes catalog, such as
> Mitotrackers.
> An anesthetized zebrafish larva or xenopus tadpole in a blob of 1-2%
> agarose will show blood flow, heart, then try reflection or
> autofluorescence.  You might find transgenics.  I can give you a list of
> labels specific for neuromast sensory organs, inner ear and nasal
> epithelium.
>
> Regards,
> Glen
> Glen MacDonald
> Core for Communication Research
> Virginia Merrill Bloedel Hearing Research Center
> Box 357923
> University of Washington
> Seattle, WA 98195-7923  USA
> [phone]
> [hidden email]
>
>
>
>
>
>
>
>
> On Jan 28, 2011, at 9:29 AM, Boswell, Carl A - (cboswell) wrote:
>
> > Primary culture of neonatal rat cardiomyocytes (or chick embryos, etc.)
> will beat spontaneously, either individually, or as sheets if dense enough.
>  GFP labeling of the right structural protein will illustrate the striations
> in the live cells.  Very bizarre to see under the microscope.
> > C
> >
> >
> > Carl A. Boswell, Ph.D.
> > Molecular and Cellular Biology
> > Univ. of Arizona
> > 520-954-7053
> > FAX [phone]
> >
> >
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of John Runions
> > Sent: Friday, January 28, 2011 10:11 AM
> > To: [hidden email]
> > Subject: Simple animal demo specimen ideas
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Dear microscopists,
> >
> > Each year at our university Science Bazaar we demonstrate our imaging
> 'prowess' and equipment to the community at large (adults and kids).  This
> event is always well received and the biggest attraction of all is usually
> the dissecting microscopes because we let the kids look at their disgusting
> fingernails and they like that.  SEMs work well because we can show head
> lice and rats tongue and... well you are probably starting to see a theme.
> >
> > The problem is giving a really good demo with the confocal microscopes -
> we show GFP labelled organelles moving around inside living cells and demos
> of 3D reconstruction but this doesn't really seem to capture the kids
> imaginations.
> >
> > Does anybody have a favorite organism or staining technique that is
> simple and effective as a demo in a situation where lots of people are
> moving through fairly quickly.
> >
> > I appreciate your suggestions.
> >
> > John
> >
> > --
> > John Runions, PhD
> > Senior Lecturer in Cell Biology
> > Oxford Brookes University
> > School of Life Sciences
> > Gipsy Lane, Oxford
> > OX3 0BP
> > UK
> >
> > 01865 483 964
> > http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!
> >
>

[Guy Cox-2]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

John,

 

        Most high school students will know about cell division, so a
metaphase cell (tubulin & DAPI), can be pretty good.   Makes a good 3D
rendering too.

 

                                                                Guy

 

Optical Imaging Techniques in Cell Biology

by Guy Cox    CRC Press / Taylor & Francis

     http://www.guycox.com/optical.htm
<http://www.guycox.com/optical.htm>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

 

Phone [phone]     Fax [phone]

             Mobile [phone]

______________________________________________

      http://www.guycox.net <http://www.guycox.net>

 

 

From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of John Runions
Sent: Saturday, 29 January 2011 4:11 AM
To: [hidden email]
Subject: Simple animal demo specimen ideas

 

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear microscopists,

Each year at our university Science Bazaar we demonstrate our imaging
'prowess' and equipment to the community at large (adults and kids).
This
event is always well received and the biggest attraction of all is
usually
the dissecting microscopes because we let the kids look at their
disgusting
fingernails and they like that.  SEMs work well because we can show head
lice and rats tongue and... well you are probably starting to see a
theme.

The problem is giving a really good demo with the confocal microscopes -
we
show GFP labelled organelles moving around inside living cells and demos
of
3D reconstruction but this doesn't really seem to capture the kids
imaginations.

Does anybody have a favorite organism or staining technique that is
simple
and effective as a demo in a situation where lots of people are moving
through fairly quickly.

I appreciate your suggestions.

John

--
John Runions, PhD
Senior Lecturer in Cell Biology
Oxford Brookes University
School of Life Sciences
Gipsy Lane, Oxford
OX3 0BP
UK

01865 483 964
http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

________________________________

No virus found in this message.
Checked by AVG - www.avg.com
Version: 10.0.1204 / Virus Database: 1435/3407 - Release Date: 01/27/11

[George McNamara]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Little kids love the pollen that looks like mickey mouse's head.
Available on Carolina Biological Supply's mixed pollen slide.

On 1/28/2011 12:26 PM, Kathryn Spencer wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I've used pollen for just this type of demo. A variety of your local flora makes it more real, and the confocal is absolutely necessary to reveal the structure, instead of being just a bright blob. Autofluorescent = no prep time. I just took some sticky tape out to some plants around the building, put mounting media on it, and covered with a coverslip. If you can show a Widefield, unresolved blob side-by-side, it is effective. Experiment beforehand to find the most interesting and diverse grains in your area, and bring in a cutting from that plant.
> Kathy
> The Scripps Research Institute
> La Jolla, CA
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of John Runions
> Sent: Friday, January 28, 2011 9:11 AM
> To: [hidden email]
> Subject: Simple animal demo specimen ideas
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear microscopists,
>
> Each year at our university Science Bazaar we demonstrate our imaging
> 'prowess' and equipment to the community at large (adults and kids).  This
> event is always well received and the biggest attraction of all is usually
> the dissecting microscopes because we let the kids look at their disgusting
> fingernails and they like that.  SEMs work well because we can show head
> lice and rats tongue and... well you are probably starting to see a theme.
>
> The problem is giving a really good demo with the confocal microscopes - we
> show GFP labelled organelles moving around inside living cells and demos of
> 3D reconstruction but this doesn't really seem to capture the kids
> imaginations.
>
> Does anybody have a favorite organism or staining technique that is simple
> and effective as a demo in a situation where lots of people are moving
> through fairly quickly.
>
> I appreciate your suggestions.
>
> John
>
>    

--


George McNamara, PhD
Analytical Imaging Core Facility
University of Miami

[Rosemary.White]

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi John,

What about some small pond creatures slowed down in the usual goop - I've
forgotten what it's called, probably based on glycerol.  Something
recognisable as an organism - pollen is good, too, as suggested, and doesn't
need to be slowed down.  Pond life usually has some autofluorescence, or can
be stained lightly, and people can compare the dissector image (often a
small swimming dot) with details on the confocal.

Cleared and stained insect heads are pretty good on the confocal too.  Some
spectacular images from groups here are the nervous systems of various
creatures - even the nematode nervous system is beautiful.  These have
various FP-expression to highlight different systems, mostly olfactory,
given the interests of our colleagues in entomology.

I've found that people are interested that chlorophyll fluoresces red - and
if you also have a GFP torch and yellow glasses and can show them red (and
GFP-green) fluorescing plants in a dark alcove, that's usually pretty
interesting.  You do sometimes get the anti-GM questions, though, and if
your regulatory environment is anything like it is here, you'd have to do it
in a PC2 facility....  Alternatively, you can show the brighter red patches
in green and white variegated leaves.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E [hidden email]


On 29/01/11 4:10 AM, "John Runions" <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear microscopists,
>
> Each year at our university Science Bazaar we demonstrate our imaging
> 'prowess' and equipment to the community at large (adults and kids).  This
> event is always well received and the biggest attraction of all is usually
> the dissecting microscopes because we let the kids look at their disgusting
> fingernails and they like that.  SEMs work well because we can show head
> lice and rats tongue and... well you are probably starting to see a theme.
>
> The problem is giving a really good demo with the confocal microscopes - we
> show GFP labelled organelles moving around inside living cells and demos of
> 3D reconstruction but this doesn't really seem to capture the kids
> imaginations.
>
> Does anybody have a favorite organism or staining technique that is simple
> and effective as a demo in a situation where lots of people are moving
> through fairly quickly.
>
> I appreciate your suggestions.
>
> John

[Christian-103]

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A three or four channel z series through a fruit fly often gives some great stuff to work with and without any staining.  Most insects heads, eyes and antennae have enough autofluorescence to wow them, and you can see the organism with the naked eye.

If you wanted to use dissecting, compound, confocal and SEM you could simply use eye lashes.  A little DAPI or PI for the confocal and nothing special for the other 'scopes might work.  I prefer to take something they've seen thousands of time and really show them what those things look like.  It makes it more real, in my opinion.

[Caroline Bass]

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Do let us know what you decide upon and how it goes with the kids. I've
been following the discussion with great interest.



On 1/28/11 12:10 PM, "John Runions" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Dear microscopists,
>
>Each year at our university Science Bazaar we demonstrate our imaging
>'prowess' and equipment to the community at large (adults and kids).  This
>event is always well received and the biggest attraction of all is usually
>the dissecting microscopes because we let the kids look at their
>disgusting
>fingernails and they like that.  SEMs work well because we can show head
>lice and rats tongue and... well you are probably starting to see a theme.
>
>The problem is giving a really good demo with the confocal microscopes -
>we
>show GFP labelled organelles moving around inside living cells and demos
>of
>3D reconstruction but this doesn't really seem to capture the kids
>imaginations.
>
>Does anybody have a favorite organism or staining technique that is simple
>and effective as a demo in a situation where lots of people are moving
>through fairly quickly.
>
>I appreciate your suggestions.
>
>John
>
>--
>John Runions, PhD
>Senior Lecturer in Cell Biology
>Oxford Brookes University
>School of Life Sciences
>Gipsy Lane, Oxford
>OX3 0BP
>UK
>
>01865 483 964
>http://www.brookes.ac.uk/lifesci/runions/HTMLpages/index.html!

[Jeffrey L. Travis]

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Many typical zoological specimens are stained such that they fluoresce.
I have used whole-mount slides of /Obelia/ polyps and tapeworm scolex
with spectacular results.  There is nothing quite like a 3-D projection
of some critter that has tentacles, or hooks and suckers to grab the
attention of young and old visitors alike.



On 1/29/2011 7:32 AM, George McNamara wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Little kids love the pollen that looks like mickey mouse's head.
> Available on Carolina Biological Supply's mixed pollen slide.
>
> On 1/28/2011 12:26 PM, Kathryn Spencer wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> I've used pollen for just this type of demo. A variety of your local
>> flora makes it more real, and the confocal is absolutely necessary to
>> reveal the structure, instead of being just a bright blob.
>> Autofluorescent = no prep time. I just took some sticky tape out to
>> some plants around the building, put mounting media on it, and
>> covered with a coverslip. If you can show a Widefield, unresolved
>> blob side-by-side, it is effective. Experiment beforehand to find the
>> most interesting and diverse grains in your area, and bring in a
>> cutting from that plant.
>>     Kathy
>> The Scripps Research Institute
>> La Jolla, CA
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]] On Behalf Of John Runions
>> Sent: Friday, January 28, 2011 9:11 AM
>> To: [hidden email]
>> Subject: Simple animal demo specimen ideas
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Dear microscopists,
>>
>> Each year at our university Science Bazaar we demonstrate our imaging
>> 'prowess' and equipment to the community at large (adults and kids).  
>> This
>> event is always well received and the biggest attraction of all is
>> usually
>> the dissecting microscopes because we let the kids look at their
>> disgusting
>> fingernails and they like that.  SEMs work well because we can show head
>> lice and rats tongue and... well you are probably starting to see a
>> theme.
>>
>> The problem is giving a really good demo with the confocal
>> microscopes - we
>> show GFP labelled organelles moving around inside living cells and
>> demos of
>> 3D reconstruction but this doesn't really seem to capture the kids
>> imaginations.
>>
>> Does anybody have a favorite organism or staining technique that is
>> simple
>> and effective as a demo in a situation where lots of people are moving
>> through fairly quickly.
>>
>> I appreciate your suggestions.
>>
>> John
>>
>
>

[Jurkevic, Aleksandr]

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Hello,

Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?


Aleksandr Jurkevic, PhD
Associate Director
Molecular Cytology Core
University of Missouri-Columbia
120 Life Science Center
1201 E. Rollins St.
Columbia, MO 65211-7310

[George McNamara]

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Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a
25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses
to 2x. NA is what matters.

On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello,
>
> Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?
>
>
> Aleksandr Jurkevic, PhD
> Associate Director
> Molecular Cytology Core
> University of Missouri-Columbia
> 120 Life Science Center
> 1201 E. Rollins St.
> Columbia, MO 65211-7310
>
>    

--


George McNamara, PhD
Analytical Imaging Core Facility
University of Miami

[Martin Seem]

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Hi Aleksandr,

we were told the same thing before buying our Leica SP5 confocal
microscope about 4 years ago. Hopefully there will be sufficient demand
for such an objective that Leica decides to produce it.

However, the company was very service minded and delivered (on our
request) our new confocal equipped with a Zeiss 40x 1.2 NA water
objective that they fitted to the turret of the DMI6000 by using an
adapter. The solution works quite well from a usability standpoint but I
believe the tests performed by Leica, if my memory serves me correctly,
showed that chromatic aberration was not fully corrected making it less
ideal for multichannel imaging.

Martin


-------------------------------
Martin Seem
Graduate student
Department of biology
NTNU
-------------------------------


On 25/02/2011 16:10, Jurkevic, Aleksandr wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hello,
>
> Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?
>
>
> Aleksandr Jurkevic, PhD
> Associate Director
> Molecular Cytology Core
> University of Missouri-Columbia
> 120 Life Science Center
> 1201 E. Rollins St.
> Columbia, MO 65211-7310

[Craig Brideau]

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Some manufacturers partly compensate their objectives in the tube lens
inside the microscope.  If you use other brands of objectives with such a
microscope you will have poorer performance:  The off-brand objective will
not be designed to work with the internal tube lens.

Craig


On Mon, Feb 28, 2011 at 6:16 AM, Martin Seem <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Aleksandr,
>
> we were told the same thing before buying our Leica SP5 confocal microscope
> about 4 years ago. Hopefully there will be sufficient demand for such an
> objective that Leica decides to produce it.
>
> However, the company was very service minded and delivered (on our request)
> our new confocal equipped with a Zeiss 40x 1.2 NA water objective that they
> fitted to the turret of the DMI6000 by using an adapter. The solution works
> quite well from a usability standpoint but I believe the tests performed by
> Leica, if my memory serves me correctly, showed that chromatic aberration
> was not fully corrected making it less ideal for multichannel imaging.
>
> Martin
>
>
> -------------------------------
> Martin Seem
> Graduate student
> Department of biology
> NTNU
> -------------------------------
>
>
>
> On 25/02/2011 16:10, Jurkevic, Aleksandr wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hello,
>>
>> Leica doesn't currently have a 40x water immersion objective although I
>> heard that they have one in the pipeline. Has anyone used a 40x water from a
>> different manufacturer with success on this frame?
>>
>>
>> Aleksandr Jurkevic, PhD
>> Associate Director
>> Molecular Cytology Core
>> University of Missouri-Columbia
>> 120 Life Science Center
>> 1201 E. Rollins St.
>> Columbia, MO 65211-7310
>>
>

[Stephen Cody-2]

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G'day Aleksandr,

Do you require this lens for confocal and or multiphoton microscopy? If so you should follow George's advice.

If you purchase a high NA 20x (or 25x) lens. Then this will perform perfectly at zoom 1 with a wide field of view, and as described be George zoom in 2 or 3 times and have the equivalent performance of a 40 or 60x. Saving you the cost of several lenses.

If however if you need a 40x for conventional microscopy, then you may need a 40x.

Mismatching objectives from manufactures may be possible, but is not a good idea, especially if you want to use confocal microscopy to optically section deep within tissue. The loss of chromatic correction I think would result in very significant loss of signal strength as you go deeper into tissue with confocal. Maybe not so problematic if using for conventional microscopy or multiphoton with NDDs.

I wouldn't mix manufactures lenses on confocal if you were hoping for good depth penetration. Which is likely the reason why you want a water immersion lens in the first place.

Cheers Steve

Stephen H. Cody

On 26/02/2011, at 11:43 AM, George McNamara <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a 25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses to 2x. NA is what matters.
>
> On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hello,
>>
>> Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?
>>
>>
>> Aleksandr Jurkevic, PhD
>> Associate Director
>> Molecular Cytology Core
>> University of Missouri-Columbia
>> 120 Life Science Center
>> 1201 E. Rollins St.
>> Columbia, MO 65211-7310
>>
>>  
>
>
> --
>
>
> George McNamara, PhD
> Analytical Imaging Core Facility
> University of Miami

[G. Esteban Fernandez]

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Steve,

Just to clarify, you think the loss of resolution/signal is more with
chromatic aberration than by reducing the NA by ~0.2 plus air/water RI
mismatch?

Thanks,
Esteban

On Fri, Mar 4, 2011 at 2:46 PM, Stephen Cody <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> G'day Aleksandr,
>
> Do you require this lens for confocal and or multiphoton microscopy? If so
> you should follow George's advice.
>
> If you purchase a high NA 20x (or 25x) lens. Then this will perform
> perfectly at zoom 1 with a wide field of view, and as described be George
> zoom in 2 or 3 times and have the equivalent performance of a 40 or 60x.
> Saving you the cost of several lenses.
>
> If however if you need a 40x for conventional microscopy, then you may need
> a 40x.
>
> Mismatching objectives from manufactures may be possible, but is not a good
> idea, especially if you want to use confocal microscopy to optically section
> deep within tissue. The loss of chromatic correction I think would result in
> very significant loss of signal strength as you go deeper into tissue with
> confocal. Maybe not so problematic if using for conventional microscopy or
> multiphoton with NDDs.
>
> I wouldn't mix manufactures lenses on confocal if you were hoping for good
> depth penetration. Which is likely the reason why you want a water immersion
> lens in the first place.
>
> Cheers Steve
>
> Stephen H. Cody
>
> On 26/02/2011, at 11:43 AM, George McNamara <[hidden email]>
> wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a
> 25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses to
> 2x. NA is what matters.
> >
> > On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
> >> *****
> >> To join, leave or search the confocal microscopy listserv, go to:
> >> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> >> *****
> >>
> >> Hello,
> >>
> >> Leica doesn't currently have a 40x water immersion objective although I
> heard that they have one in the pipeline. Has anyone used a 40x water from a
> different manufacturer with success on this frame?
> >>
> >>
> >> Aleksandr Jurkevic, PhD
> >> Associate Director
> >> Molecular Cytology Core
> >> University of Missouri-Columbia
> >> 120 Life Science Center
> >> 1201 E. Rollins St.
> >> Columbia, MO 65211-7310
> >>
> >>
> >
> >
> > --
> >
> >
> > George McNamara, PhD
> > Analytical Imaging Core Facility
> > University of Miami
>

[Jurkevic, Aleksandr]

*****
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Steve,

The  objective I was asking is intended for confocal microscopy. I agree with yours and George's arguments that without a genuine Leica 40x WI objective on the market, our best option would be to zoom with a 20x or 25x WI Leica objective. I am not sure if one can achieve the performance of a 63x WI just by zooming with a 20x WI objective that has lower NA.

I appreciate all responses to my posting.

Aleksandr  

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Stephen Cody
Sent: Friday, March 04, 2011 4:47 PM
To: [hidden email]
Subject: Re: 40x water immersion objective on Leica miscroscope

*****
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G'day Aleksandr,

Do you require this lens for confocal and or multiphoton microscopy? If so you should follow George's advice.

If you purchase a high NA 20x (or 25x) lens. Then this will perform perfectly at zoom 1 with a wide field of view, and as described be George zoom in 2 or 3 times and have the equivalent performance of a 40 or 60x. Saving you the cost of several lenses.

If however if you need a 40x for conventional microscopy, then you may need a 40x.

Mismatching objectives from manufactures may be possible, but is not a good idea, especially if you want to use confocal microscopy to optically section deep within tissue. The loss of chromatic correction I think would result in very significant loss of signal strength as you go deeper into tissue with confocal. Maybe not so problematic if using for conventional microscopy or multiphoton with NDDs.

I wouldn't mix manufactures lenses on confocal if you were hoping for good depth penetration. Which is likely the reason why you want a water immersion lens in the first place.

Cheers Steve

Stephen H. Cody

On 26/02/2011, at 11:43 AM, George McNamara <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Leica has a 20x/1.0 NA objective lens, and, if I recall correctly, a 25x/0.95NA - if this is on a laser scanning confocal, zoom these lenses to 2x. NA is what matters.
>
> On 2/25/2011 10:10 AM, Jurkevic, Aleksandr wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hello,
>>
>> Leica doesn't currently have a 40x water immersion objective although I heard that they have one in the pipeline. Has anyone used a 40x water from a different manufacturer with success on this frame?
>>
>>
>> Aleksandr Jurkevic, PhD
>> Associate Director
>> Molecular Cytology Core
>> University of Missouri-Columbia
>> 120 Life Science Center
>> 1201 E. Rollins St.
>> Columbia, MO 65211-7310
>>
>>  
>
>
> --
>
>
> George McNamara, PhD
> Analytical Imaging Core Facility
> University of Miami