I need to prepare my biological samples for SEM and I don't have this buffer. Is there any chance I can skip this step of using cacodylate buffer?
I asked the question why using an other poisonous and carcinogenic buffer if the results are the same with PBS.
I found this:
"Author: ProSciTech Pty Ltd
Last Updated: 2014-05-12
Sodium cacodylate trihydrate is the most favoured buffering vehicle used with fixatives in specimen preparation for electron microscopy. Also called cacodylate or cacodylic acid, this chemical avoids the microprecipitation on thin sections that can occur with phosphate buffers if the specimen is not well rinsed between pre- and post-fixation. The chemical does not form a precipitate in sea water. Ultrastructural preservation is excellent."
(http://laboratoryresource.com.au/?navaction=getitem&id=45)
So it depends how sadisfying your results will be.
Hi,
I faced the same issue and yes you can avoid cacodylate buffer. Glutaraldehyde can be diluted in PBS.
Thank you your response, but did you had the same good results?
Yes, It did not affect the quality of the samples preparation, but progressive dehydration and final drying are still important to get good samples.
I second that suggestion. PBS is a good substitute for cacodylate buffer for SEM.
I asked the question why using an other poisonous and carcinogenic buffer if the results are the same with PBS.
I found this:
"Author: ProSciTech Pty Ltd
Last Updated: 2014-05-12
Sodium cacodylate trihydrate is the most favoured buffering vehicle used with fixatives in specimen preparation for electron microscopy. Also called cacodylate or cacodylic acid, this chemical avoids the microprecipitation on thin sections that can occur with phosphate buffers if the specimen is not well rinsed between pre- and post-fixation. The chemical does not form a precipitate in sea water. Ultrastructural preservation is excellent."
(http://laboratoryresource.com.au/?navaction=getitem&id=45)
So it depends how sadisfying your results will be.
I agree with Bing Ham...to greatly preserve ultra-structural details for electron microscopy sodium cacodylate buffer appears to be my best option. That is not the only determiner though, you might want to carefully do your dehydration steps too.
you can use another method such fix samples with deposit that on to clean glass slides and dry it. But its can`t give you clearly image so I suggested to use 2.5% (wt/vol) glutaraldehyde in 0.05 M sodium cacodylate buffer for 2 hr, and washed three times for 5 min each with the cacodylate buffer to minimized structure changes.
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