Studying Neurons with Electron Microscopy (FIB-SEM)

Contacts:

Kristian Mølhave, DTU-Nanotech, 2512 6672 (kristian.molhave@nanotech.dtu.dk)

http://www.nanotech.dtu.dk/Medarbejdere/Staff/Molhave_Kristian.aspx

SEM Image of dried cells grown on an an electron transparent Si-nitride thin film membrane. The cells are imaged through the 100nm membrane and a FIB-SEM cut into on of the cells to see the 3D structure of it

Introduction: We are developing a method to construct 3D nanoscale resolution images of cells and how they grown on other structures such as nanostructured surfaces that turn out to influence their behaviour in many ways. We will use SEM combined with focused ion beam milling (FIB-SEM) at the new Center for Electron Nanoscopy to cut slices into epoxy embedded cells grown on nanostructures

Project goal: This project is about making a series of SEM images as the cell is cut slice-by-slice in the FIB-SEM, that enables 3D images to be reconstructed of the cell.We will use the FIB-SEM systems at the new Center for Electron Nanoscopy (CEN).

Description: In this cross disciplinary project you will need to prepare nanostructured surfaces - either use some that we have prepared or you can help making the chips in the cleanroom. We make these nanostructures on a very thin silicon nitride membrane (50-100nm) that is transparent to the electron beam, so we can image the cells directly through the membrane in the microscope.

Then we will teach you how to culture cells, so they attach to the nanostructures, and later fix and stain the cells with chemical compounds to make them ready to be observed in electron microscopes.

Finally you will learn about electron microscopy and focused ion beam milling so you can make the image series and optimize the whole process.

Perspectives: Artificial nanostructured surfaces have been shown to influence cell behaviour in many ways and this is a new research area several groups at DTU are working on. We are for instance  studying a special kind of neuron-like cells. The methods developed in this project will be a basic tool for such investigations in the future.