Abbelight
Caterina Severi
Workshop Room 7
TIRF (Total Internal Reflection Fluorescence) microscopy is often used in biology for studying the cell membrane. This technique exploits the total internal reflection of the light beam on the microscope slide, which induces the propagation of an evanescent wave through the first hundreds of nanometers of the sample, allowing the excitation of only a very thin section of the sample close to the surface
Because of its optical section of few hundreds of nanometers, TIRF microscopy allows a higher SNR comparing to classical epifluorescence microscopy techniques. In operation with ultra-fast sCMOS or EMCCD cameras, it provides a good temporal sampling which can be combined with STORM (Stochastic Optical Reconstruction Microscopy) and PALM (Photo-Activated Localization Microscopy) super-resolution techniques for an optimal spatial and temporal dynamic range.
In this workshop, you can discover our TIRF product: a hardware and software solution that provides an optically perfect TIRF imaging. Adaptable to any combination of microscopes, objectives and third parts, our product guarantees the automatically calibrated and reproducible position of the TIRF angles. Coupled with a multi-color optical module, it offers an optimal and simultaneous imaging of biological structures with the benefits of our ultra widefield ASTER technology.
Schedule :
10 min : introduction to TIRF microscopy
15 min : demonstration of our 4 channels detection module alignment
10 min : Simultaneous multi-color TIRF imaging on cells
15 min : demonstration of the automatic calibration TIRF angles software
10 min : summary and Q&A
Figure 1 : COS 7 cells (actin labelled with phalloidin-488 and mitochondria labelled with AF-647)