Electron Microscopy for 3D reconstructions of serial sections – image and post-image processing course
TEM is generally believed to be limited to 2D images. However, a large amount of detail can be obtained when using serial TEM images. It is possible to reconstruct 3D structures from those images, giving detailed information on volume and area as well as more information on morphology.
The size (or depth) of the final 3D structure reconstructed from serial images will depend on how many images there are in a series. Using a standard TEM, we generally use 100 – 150 images per series to obtain plenty of detail about the structures.
This is a 2-day course that will teach you how to perform 3D reconstruction of a series of TEM images acquired with a standard TEM. The skills you will learn include:
- Image acquisition from series of images,
- Montaging and alignment of series of images
- Reconstruction of the selected structure
We will demonstrate you the necessary steps involved in TEM imaging and processing the images. You will then repeat each step until you feel comfortable with it so you can continue learning the next one. You will also have some time to practice obtained knowledge using an exercise, under our supervision.
At the end of the course you should be comfortable obtaining the series of TEM images and using various software packages to create a 3D reconstruction of a region of choice.
At the course completion you will receive a certificate of attendance.
In addition to the course time stated in the table there will be lunch and 2 coffee breaks provided, adding 1 h 40 min to the course time of each day. The total time at the course will therefore be 6 h 40 min each day.
|Alignment necessary for serial imaging
|Image acquisition on serial sections, operating and setting up SerialEM software.
|Practise of setting up SerialEM software for imaging
|Handling images – 8-bit transform, aberration correction, montaging, alignment + practise on another set of images
|Total time (split in 2 days)