I have been working with rice leaf samples for Transmission electron microscopy. We are using spurr resin for embedding. However, while trimming of the resin during ultramicrotomy, the rice leaf samples are coming out intact as soon as the resin is trimmed, making it very difficult to cut ultrathin sections. The leaf sample seems very hard even after all the processing steps. Any insight into the possible reasons for this problem?
I guess the Spurr resin is harder than your sample.
I got this from their website:
The introduction of Spurr1 resin provided the microscopist with a resin of exceptional penetration qualities. Unlike the other epoxies, Spurr resin with a viscosity of 60 cps will readily penetrate membrane walls, hard tissue, rocks, and other minerals.
It is more suited for woody material. I think you should use another resin or embedding material of similar strength as the plant material for embedding or no resin at all. Maybe just slice it with a hand microtome?
Hello,
The resin or one of its components did not get inside the leaf (the different components of the resin do not have the same viscosity). So no polymerization inside while outside resin correctly polymerizes. Your leaf is not embedded.
How long do you infiltrate the leaf with increasing concentration of resin versus solvent? Because of cell wall, it should be several days compared to animal tissues. In addition leaf pieces should be small.
Spurr is less viscous than Epon so it should be the best choice to get it inside the leaf.
Do you use the accelerator DMP30 in your Spurr? You should try BDMA that is much less viscous than DMP30. If DMP30 is not going inside the leaf, no polymerization.
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