Integrated Super-Resolution Fluorescence Microscope in a TEM
In this project, we will develop and apply methods and techniques to combine high resolution resolution light microscopy with transmission electron microscopy (TEM). Three groups will work closely together focusing on different aspects. The Koster team will focus on the development of super-resolution correlative microscopy approaches in general, and their implementation and optimization for integrated light and transmission electron microscopy in particular. The Gerritsen team will focus on the technical realization and characterization of a concise super-resolution optical system that will be built in a TEM and that is based on a the commercial iCorr module by FEI company. The Klumperman team will focus on the development of optimal specimen preparation procedures for integrated light and electron microscopy for the Koster and Gerritsen teams.
Integration of a 3D-Confocal Fluorescence Microscope in a FIB-SEM
This project is focused on the development of 3-D correlative microscopy using a dual beam system. The commercial dual beam FIB/SEM is capable of producing 3-D high resolution images by repetitively removing thin layers by Gallium ion-beam milling and recording SEM images, also known as “slice and view.” The resolution of only a few nanometres, large data sets (tens to hundreds of gigabytes) are generated that require long measurement times (several days) even for relatively small volumes.
We will integrate a confocal fluorescence microscope in the dual beam system (“3D CLEM”) to rapidly locate regions of interest in 3-D. This will result in a significantly sped-up the 3-D correlative imaging work flow.
- “Fluorescently Labeled Silica Coated Gold Nanoparticles as Fiducials for Correlative Microscopy“, (Jantina Fokkema, poster, P1-D/9, Monday)
- “Simultaneous STED-FLIM Imaging of Actin and EGFR” (Hans Gerritsen, talk, TU-AF1-PAR-B, Tuesday at 14:20)
- “Looking through a Mismatched Sample: When the Emission and Excitation PSF Are No Longer the Same” (Sergey Loginov, talk, WE-MO2-PAR-C, Wednesday at 12:00)
In addition, may I wholeheartly recommend a talk by one of our colleagues from the Soft Condensed Matter Group:
- “Single Sample Calibration and Alignment of Clsm and (3D) STED” (Ernest van der Wee, talk, WE-MO2-PAR-A, Wednesday at 12:20)