We use TEM Tomography for cellular organelles
1) Cells cultured on Quantifoil grids.
2) Tissue embedded in plastic resin.
3) Plasma membrane replicated by platinum/carbon.
1) General alignments.
2) Adjust auto eucentric height & auto focus at optimised position.
3) Calibrate the sample holder.
4) Calibrate the dynamic focus for STEM-tomography.
5) Prepare the acquisition positions for cryo-tomography.
6) Prepare step conditions before starting a tilt series.
7) Acquire tilt series.
8) Reconstruct live tilt series.
9) 3D reconstruct by IMOD or FEI Inspect3D & AMIRA.
Tilted series of Actin-based membrane skeleton with pits & caveolae in normal kidney – video: