RE: SEM inquiry

From:Philip Oshel

The staining won't affect the SEM imaging. Any embedding medium must 
be removed and the samples dried. If there is no embedding medium, 
and the samples are dry, then they're ready for coating. This could 
be the hang-up. If the samples must be left unaltered, then they 
cannot be coated. This means a field-emission SEM operating at low 
voltage (<5kV, likely around 1 - 2 kV) may be necessary. Although I 
have photographed fresh plant leaves uncoated at 10kV. It's best to 
coat (gold-coat or gold-palladium in a sputter coater) however. This 
coating can only be removed with effort and the use of methods that 
might at best destain the samples. Also, depending on the thickness 
needed, it might hide the staining, and it certainly will affect the 
colors of the stains. The coating is gray to gold, depending on 
thickness and surface texture. It is also somewhat transparent to 
opaque (if thick enough).
The size of the samples and whatever they're mounted on could also be 
a problem, but this depends on the stage.
If there is a chance of ruining things, then perhaps it's best to 
wait for your SEM person to return. Keep the samples dry and 
dust-free, and ignore them until then.


>It's a dog femur samples implanted with IM rods coated with Titanium foam.
>The slides were grind and polished up to 60 microns and stained with
>Stevenel's Blue/ Van gieson Picrofuchsin. We originally measure the porosity
>using Bioquant Image analysis and would like to compare the porosity using
>SEM, but we have stained all the slides. Yes!! we do have an expert on SEM,
>but he is on vacation...want to be safe... I can't go back and repeat all
>-----Original Message-----
>From: Philip Oshel []
>Sent: Thursday, August 15, 2002 3:15 PM
>To: Dimaano, Nena
>Subject: Re: SEM inquiry
>Any longer reply will require more information about the slides,
>preparation, what prep equipment you have access to, what you're
>after, what can be done to the slides, and so on.
>But you have access to lots of SEMs in your area, both commercial and
>university, and personnel to run them and prepare samples.
>>To all SEM experts:
>>Can you do SEM on a stained slides?
>>Nena Dimaano,MT/HT(ASCP)
>>Advanced Technology- Preclinical and Biological Affairs
>>Stryker Howmedica Osteonics
>>300 Commerce Court
>>Mahwah, NJ 07430
>>tel: 201-831-5338
>>fax: 201-831-6224
>Philip Oshel
>Supervisor, BBPIC microscopy facility
>Department of Animal Sciences
>University of Wisconsin
>1675 Observatory Drive

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