Labs for graduate level microscopy course?

[Kurt Thorn]

Hi All -

I'm putting together some labs for a graduate level microscopy course
I'll be teaching in the next few months and I was wondering if people
here had ideas or materials for confocal microscopy labs that they could
share.  I'm particularly interested in doing 1 or 2 3 labs comparing
widefield / laser-scanning confocal / spinning-disk confocal, with
respect to both sensitivity and rejection of out-of-focus light.  Labs
are 3 hours long and targeted and mid-level graduate students who may or
may not have microscopy experience.

I've got some ideas on how to do some of this but I'm curious to hear of
exercises that have worked well for you and would love write ups or
other materials you could share.

Thanks!
Kurt

[Paul Herzmark]

Steve Ruzin at UC Berkeley has a good one for rejection of out of focus light.

Focus on the stomata on a leaf epidermis. There are lots of fluorescent chloroplasts in the guard cells surrounding the stomata. The confocal will block the fluorescence from the deeper mesophyl layer and all the chloroplasts there.



Paul Herzmark
Specialist
[hidden email]

Department of Molecular and Cell Biology
479 Life Science Addition
University of California, Berkeley
Berkeley, CA  94720-3200
[phone]
[phone] fax

On Tue, Mar 30, 2010 at 1:30 PM, Kurt Thorn <[hidden email]> wrote:

Hi All -

I'm putting together some labs for a graduate level microscopy course I'll be teaching in the next few months and I was wondering if people here had ideas or materials for confocal microscopy labs that they could share.  I'm particularly interested in doing 1 or 2 3 labs comparing widefield / laser-scanning confocal / spinning-disk confocal, with respect to both sensitivity and rejection of out-of-focus light.  Labs are 3 hours long and targeted and mid-level graduate students who may or may not have microscopy experience.

I've got some ideas on how to do some of this but I'm curious to hear of exercises that have worked well for you and would love write ups or other materials you could share.

Thanks!
Kurt

[mmodel]

Kurt,

I suggest, keep it simple - one little step at a time. If your students haven't done microscopy before you may be surprised what they do NOT know...

Best wishes -

Mike

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kurt Thorn
Sent: Tuesday, March 30, 2010 4:31 PM
To: [hidden email]
Subject: Labs for graduate level microscopy course?

Hi All -

I'm putting together some labs for a graduate level microscopy course
I'll be teaching in the next few months and I was wondering if people
here had ideas or materials for confocal microscopy labs that they could
share.  I'm particularly interested in doing 1 or 2 3 labs comparing
widefield / laser-scanning confocal / spinning-disk confocal, with
respect to both sensitivity and rejection of out-of-focus light.  Labs
are 3 hours long and targeted and mid-level graduate students who may or
may not have microscopy experience.

I've got some ideas on how to do some of this but I'm curious to hear of
exercises that have worked well for you and would love write ups or
other materials you could share.

Thanks!
Kurt

[Patrick Van Oostveldt]

Dear,

Plant material is easy to handle.
Fruits like apple of cherry tomato's can be images as whole. The  
epidermal cells contain fluorescent chloroplasts and the tha vacuoles  
are frequently filled with antocyaan dyes.
Pollen grains are always succesfull and can be obtained from lily or dafodill

Bye

Patrick


Also in reflection they generate nice images of surface.

Patrick

Quoting "Paul Herzmark" <[hidden email]>:

> Steve Ruzin at UC Berkeley has a good one for rejection of out of focus
> light.
>
> Focus on the stomata on a leaf epidermis. There are lots of fluorescent
> chloroplasts in the guard cells surrounding the stomata. The confocal will
> block the fluorescence from the deeper mesophyl layer and all the
> chloroplasts there.
>
>
>
> Paul Herzmark
> Specialist
> [hidden email]
>
> Department of Molecular and Cell Biology
> 479 Life Science Addition
> University of California, Berkeley
> Berkeley, CA  94720-3200
> [phone]
> [phone] fax
>
>
> On Tue, Mar 30, 2010 at 1:30 PM, Kurt Thorn <[hidden email]> wrote:
>
>> Hi All -
>>
>> I'm putting together some labs for a graduate level microscopy course I'll
>> be teaching in the next few months and I was wondering if people here had
>> ideas or materials for confocal microscopy labs that they could share.  I'm
>> particularly interested in doing 1 or 2 3 labs comparing widefield /
>> laser-scanning confocal / spinning-disk confocal, with respect to both
>> sensitivity and rejection of out-of-focus light.  Labs are 3 hours long and
>> targeted and mid-level graduate students who may or may not have microscopy
>> experience.
>>
>> I've got some ideas on how to do some of this but I'm curious to hear of
>> exercises that have worked well for you and would love write ups or other
>> materials you could share.
>>
>> Thanks!
>> Kurt
>>
>

--
Dep. Moleculaire Biotechnologie
Coupure links 653
B 9000 GENT

tel [phone]
fax [phone]

[gradice]

Douglas Murphy's book "Fundamentals of Light Microscopy and Electronic  
Imaging" has some exercises and demos that I have used. Among these he  
includes a "build your own confocal" demo that, although I have not  
used it myself, would be a lot of fun and illustrate the pinhole effect.


On Mar 30, 2010, at 4:30 PM, Kurt Thorn wrote:

> Hi All -
>
> I'm putting together some labs for a graduate level microscopy  
> course I'll be teaching in the next few months and I was wondering  
> if people here had ideas or materials for confocal microscopy labs  
> that they could share.

Gary Radice
Department of Biology
University of Richmond
Richmond VA 23173
804-289-8107

[Littlejohn, George]

Hi Gary and list,

On a similar note, Jim Haseloff's cheaposcope pages (not to mention the rest of his pages) must be well worth a look for eager microscopists!

http://www.plantsci.cam.ac.uk/Haseloff/imaging/cheaposcope/cheaposcope.htm

This is, of course, not a confocal microscope. But is great fun!

George Littlejohn
School of Biosciences,
University of Exeter,
Mezzanine Laboratory,
Geoffrey Pope Building,
Stocker Road,
EX4 4QD, UK
******************************
Tel:  [phone] (Lab.)
       [phone] (Office)
Fax:  [phone]
E-mail:  [hidden email]

On 31 Mar 2010, at 17:17, gradice wrote:

Douglas Murphy's book "Fundamentals of Light Microscopy and Electronic  
Imaging" has some exercises and demos that I have used. Among these he  
includes a "build your own confocal" demo that, although I have not  
used it myself, would be a lot of fun and illustrate the pinhole effect.


On Mar 30, 2010, at 4:30 PM, Kurt Thorn wrote:

Hi All -

I'm putting together some labs for a graduate level microscopy  

course I'll be teaching in the next few months and I was wondering  

if people here had ideas or materials for confocal microscopy labs  

that they could share.

Gary Radice
Department of Biology
University of Richmond
Richmond VA 23173
804-289-8107