Learn & Share: TIRF Microscopy
Total internal reflection fluorescence microscopy (TIRFM) is a highly sensitive technique to perform functional investigations in living cells. The high signal to noise ratio and a resolution in z direction of usually 70-250 nm above the coverslip/water interface allows to visualize and to analyse vesicles transport and signalling events, as well as kinetic studies and single molecules detection. Physical background is the total reflection of a laser beam at the interface of glass and water and the resulting electromagnetic wave, the so-called evanescent field. The energy of the evanescent field decreases exponentially with the distance of the interface coverslip/water and allows exciting fluorochromes.
Where to go? Cellular Migration requires coordinated Transitions of Actin Cortex
Plants, Bacteria, and Fungi possess a rigid cell wall that protects the cell and gives it shape. Animal cells, such as mammalian cells, have no outer wall, which exposes their plasma membrane to the…Read article
TIRF Publication List
This monthly updated references list presents current papers using Leica AM TIRF in the major application fields for TIRF microscopy.Read article
Co-Orientation: Quantifying Simultaneous Co-Localization and Orientational Alignment of Filaments in Light Microscopy
Co-localization analysis is a widely used tool to seek evidence for functional interactions between molecules in different color channels in microscopic images. Here we extend the basic…Read article
Video Talk by Daniel Axelrod: Total Internal Reflection Fluorescence (TIRF) Microscopy
Total Internal Reflection Fluorescence (TIRF) Microscopy is a technique that only illuminates dye molecules near a surface. In this video, the pioneer of TIRF Microscopy describes what this technique…Read article
Video Interview with William Hughes
William Hughes works at the Garvan Institute of Medical Research, Sydney (Australia). In his Lab Head position he is interested in the causes of diabetes particularly looking at changes in exocytic…Read article
Controlling the TIRF Penetration Depth is Mandatory for Reproducible Results
The main feature of total internal reflection fluorescence (TIRF) microscopy is the employment of an evanescent wave for the excitation of fluorophores instead of using direct light. A property of the…Read article
Total Internal Reflection Fluorescence (TIRF) Microscopy
Total internal reflection fluorescence (TIRF) is a special technique in fluorescence microscopy developed by Daniel Axelrod at the University of Michigan, Ann Arbor in the early 1980s. TIRF microscopy…Read article
Applications of TIRF Microscopy in Life Science Research
The special feature of TIRF microscopy is the employment of an evanescent field for fluorophore excitation. Unlike standard widefield fluorescence illumination procedures with arc lamps, LEDs or…Read article
Protein Transport Processes at the Apical Membrane of Polarized Epithelial Cells
Due to their special role in organ function and the exchange of biological components some body cells developed certain polarization characteristics. These are reflected in differences of their plasma…Read article
The New Repository on the Block
The need for data validation and accessibility has never been greater than it is today. We are inundated with information from a multitude of resources, but how can we easily evaluate the accuracy of…Read article
TIRF Microscopy of the Apical Membrane of Polarized Epithelial Cells
Application of TIRF microscopy (Total Internal Reflection Fluorescence) allows the visualization of structures at the apical surface of polarized epithelial cells that have been hidden in conventional…Read article
Exploring Cell Logistics
Using TIRF microscopy, scientists have been able to take a closer look at intracellular transport processes with the example of the galactose-binding protein Galectin-3, which has been identified as a…Read article