Re: Practice samples

Posted by Rosemary.White on
URL: http://confocal-microscopy-list.275.s1.nabble.com/ATTO647N-and-FCS-tp1499210p1507227.html

Hi Jon,
As Tobias says, plants are great for their variety of autofluorescence.  I
cut hand sections (from a couple of bushes just outside the building) in
front of school groups and put them straight under the fluorescence or
confocal microscope.  UV/405nm illumination will give a big variety of
fluorescence from cell walls and cell contents.  There are also cheap stains
like rhodamine B, berberine sulfate, Schiff's reagent, calcofluor white (the
brightener in washing powder) that you can use, which mainly stain cell
walls.
Another cool thing to do is get a cube of either uranium glass or
fluorescent plastic and put this under the objective to show the light path
and explain the focal plane - or fill a cuvette with fluorescent solution.
Does a much better job than diagrams on a piece of paper!
cheers,
Rosemary

On 15/11/08 5:56 PM, "Tobias Baskin" <[hidden email]> wrote:

> Jon,
>         Plants are full of autofluorescence. If you cut a section (a
> careful hand section is fine) of a stem or root, most times you will
> see lovely fluorescence. What is nice about this is that you can have
> students collect their own matieral, try different places on the
> plant, aged material, etc etc. It takes a little knack to cut nice
> smooth hand sections so they coverslip up nice and show good images
> near the coverslip. You don't have to stain at all, just mount and
> observe. There can also be nice differences at different wavelengths.
>         Have fun,
>                 Tobias
>
>
> At 8:45 AM -0800 11/14/08, Jonathan M Krupp wrote:
>> Hi:
>>
>> This is a pretty basic and simple question. I need some advice about
>> rounding up some samples to use in an elementary light microscopy
>> class that includes basic fluorescence.
>>
>> I am at a community college and my background is in EM and
>> brightfield LM. I have a passing familiarity with confocal et al but
>> not enough to know much. I maintained a confocal, but never ran
>> experiments or prepared much in the way of samples. Now I have to
>> get something together to demonstrate the fundamentals of
>> fluorescence to students in a new job.
>>
>> I have been relying on chlorophyll autofluorescence up til now, but
>> would like to add anything that would be easy to do. We have a
>> simple scope with filters for FITC, rhodamine, and DAPI, I think.
>> What would be some fool proof, easy to get samples to try?
>>
>> In addition, I would eventually like to add some kind of confocal
>> experience to this class, any ideas about where to find an
>> inexpensive system would be great.
>>
>> Thanks
>>
>> Jon
>> San Joaquin Delta College
>> Stockton, CA 95207
>> [hidden email]
>
>
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