- Unraveling Intermediate Filaments
- The super resolution solution
- Award date
- 24 February 2017
- Number of pages
- Document type
- PhD thesis
- Faculty of Science (FNWI)
- Swammerdam Institute for Life Sciences (SILS)
Intermediate Filaments (IFs) carry out major functions in cells. Several diseases have been associated with malfunctioning IFs in the cells and among them are certain sub types of cancer. To determine the structure and organization of IFs, we have used Single Molecule Localization Microscopy (SMLM) . We have shown that keratin IF plays a key role in Hemidesmosomes (HDs) organization in cultured keratinocytes.
In addition, we have studied the orientational alignment between vimentin, another abundant IF protein, and MT. With SR, we have observed spatial proximity between single vimentin and MT filaments and we have quantitatively shown that interaction between vimentin and MT are cell-type dependent.
Furthermore, we have addressed several critical issues in SMLM imaging. First, we have developed a new buffer which supports multi-color imaging. OxEA, an Oxyrase based imaging buffer, elevates localization precision for commonly used dyes and enhances blinking of other dyes. Second we have improved a dedicated chamber for SR imaging, so called Oxygen Tight Chamber (OTC). In the OTC, we have minimized drift by introducing an alternative curing procedure that reduces stress in the bond between glass and plastic.
In brief, in this thesis we have optimized imaging condition and post processing analysis for SMLM microscopy. With the resulting high quality SR images and precise quantification methods, we have contributed to the biological knowledge in the IF field.
Thesis (complete) (Embargo until 24 February 2019)
Chapter 4: Studies of keratin filament structure at super resolution (Embargo until 24 February 2019)
- Supplementary materials
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