I have technical difficulties in using ultramicrotome to cut my hydrogel beads. However, I did read about the use of nitrogen liquid treatment and followed by cutting using razor blade.In short, I hope to obtain some suggestions on cutting a SMOOTH (not rough) cross-sectional surface for my bead samples
How big are the beads and what do you want to observe in them? try the freezing and cutting, it is a 5 minute job, then you'll know.
Agreed. There is cryostat section as an option, depending on the size of your sample.
If you changed the solvent for acohol, or acetone you can embed, and cut. If you can paint it, you can study by SEM too.
Hi sir and madam, thanks for your replies.
To Kenton Arkill, my nearly-round bead sample has an average diameter of 2.8 mm or 0.28 cm. I want to see the inner pore structures and its distributions, How did you cut your sample after freezing them? Is there any equipment you use to facilitate the cutting process?
To Branka Vulesevic , I have checked with my university's research facilities and found that cryostat is not available (not being bought yet). Thanks for your suggestion.
To Chintan Dalwadi , I think you can use CLSM diffusional studies of hydrogel. If you really need a details of morphological studies, i suggest you can use AFM.
To Jozsef Bako , my bead samples are not like a layer of cell tissues. My bead samples have higher height, length and width than tissue cells for sure. In this case, can I still fix my beads using alcohol or acetone onto a solid support? If they can be done, I can use them more easily then. Right now, i do not fix my bead sample even using simple cutting.
To Clarissa Villinger , you do understand my difficulties already (laugh). My research lab actually does have the ultramicrotome, but there is no technician around to neither prepare the diamond knife nor buy those necessary chemicals and raw materials to properly use the equipment. I am a bit disappointed actually.
Your experience in using the focused ion beam really valuable here but my research lab does not have the FIB right now. Do you have any suggestion on cutting my bead sample so fast and rapid that it provide a soft surface texture cross-sensationally? You can have a look at my SEM image which shows the rough (not smooth) cutting of cross-section bead.
To Kevin Yong, Thank you for your reply. But the problem is that i cant formulate film and for AFM film is required so i want to use CLSM and CRYO-SEM option is there but it is not available in india so
If you have an ultramicrotome, use a glass knife. Alternatively you could use a razor blade cooled liquid nitrogen to fracture the beads.
Hi Kevin
Ultratome: if you have a glass knife cutter that would suffice, but your beads are to big and would need to be cut down anyway.
I think a normal fresh razor blade would be good enough: frozen or not. Then use SEM, either still frozen (if you have the facilities). I assume you have checked your Universities EM failities:
http://www.monash.edu.au/research/infrastructure/platforms/electron.html
Somebody there will have a way... (there is a FIB SEM on the website but they probably don't have the bio embedding), try a pathology lab in big hospitals near by.
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