Scratching the Cell Surface

Most biological microscopes delve deep into the cell, imaging optical slices that can be put together into a three-dimensional rendering of what lies beneath the cell membrane.

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Adapted from Nikon Microscopy U (http://www.microscopyu.com)

TIRF microscopy uses the principle of total internal reflection to selectively activate fluorophores near the slide-sample interface while ignoring more deeply embedded ones. That makes the technique useful for probing membrane events, cell motility, and single-molecule biochemistry – anything that occurs near the slide surface.

Most biological microscopes delve deep into the cell, imaging optical slices that can be put together into a three-dimensional rendering of what lies beneath the cell membrane. But a lot of biology takes place at the cell surface. Vesicles and receptors cycle between the membrane and the cytoplasm, propagating cellular signals by way of transient interactions between cellular proteins, macromolecular assemblies, and organelles. To catch a glimpse of these events, a growing number of researchers use total internal reflection fluorescence (TIRF) microscopy, also known as evanescent wave microscopy.

Unlike confocal microscopy, which takes optical slices deep within cells at ...

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