The EML staff will perform TEM sample preparation, ultramicrotomy, post-staining and TEM imaging for your project.
We have both morphological (conventional) TEM and Correlative Light and Electron Microscopy (CLEM; as in: https://rdcu.be/d2h50) pipelines available.
Basic protocol for the TEM Preparation as follows:
https://em-lab.berkeley.edu/EML/tem-generic-protocol
Ultramicrotomy with:
The EML has Leica ultramicrotomes (UC6 and UC7), also equipped with cryo-ultramicrotomy mount, for sectioning resin blocks. 50nm – 100nm sections can be collected onto a variety of grid types as needed (slot or mesh). For projects that need correlation back to whole tissues, we employ toluidine blue staining of “thick” sections collected onto glass slides, light microscopy images are taken for reference.
Serial Sectioning can be accommodated as needed and is standard with our CLEM pipeline.
Grid post-staining with:
We employ aqueous Uranyl Acetate and Reynold’s Lead Citrate to post-stain grids for TEM contrast.
TEM imaging with:
The Tecnai 12 120kV TEM is an advanced transmission electron microscope that offers magnification ranges from 120X to 300,000X. The microscope has a lens designed to optimize imaging low contrast samples for biological or polymer material specimens.
Images are collected with a state-of-the-art Gatan Rio16 CMOS 16MB camera.
