TEM Tomography

We use TEM Tomography for cellular organelles

TEI Talos 200k VFE-TEM with FEI CMOS Detector

Samples:

1) Cells cultured on Quantifoil grids.

2) Tissue embedded in plastic resin.

3) Plasma membrane replicated by platinum/carbon.

 

 

Workflow:

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:

50R15

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