Re: Jellyfish for SEM

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From:Scott Whittaker <>
To:Rena Krol <>
Date:Wed, 11 Aug 1999 14:59:12 +0100
Content-Type:text/plain; charset="us-ascii"

Hi rena. I am new to the list, but primarily do SEM and TEM. Although I
have not actually worked with jellyfish, here is how I would proceed.

Fix in 2-2.5% glutaraldehyde in filtered seawater pH=7.6-8 (assuming
samples are marine) for 1hr. Otherwise use PBS buffer pH=7.2ish
Wash in seawater 3x 5 min.

Post-fix 1% buffered osmium tetroxide 1hr. This step is very optional
especially if you are not going to fracture the sample but the deposition
of heavy metal can help charging problems.

Wash dH2O 3x 5 min.
Graded Ethanol series 25-50-75-95-100-100% 5 min in each step.

>From here you need to either critical point dry or solvent dry with
HexaMethylDiSilizane (HMDS by far the fastest route but doesn't work for

mount, sputter, view.

If this doesn't work you may need to try some of the more exotic procedures
like OTOTO or a mercuric chloride/osmium fixation or even freezedrying from
LN2. Let me know if you need references or details on any of this. I will
be happy to pass them along. 

I have also posted this message to the microscopy listserver dealing
primarily in SEM and TEM. Good luck

At 10:37 AM 8/11/1999 -0500, you wrote:
>Hello Histonetters,
>I just received some small jellyfish (3/4 inch diameter) with attached
>Vibrio.  Does anyone have any experience with processing such a
>gelatinous critter
>for scanning electron microscopy?  Your help is appreciated!
>Rena Krol

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