Vertico spatially modulated illumination

Vertico spatially modulated illumination (Vertico-SMI) is the fastest[citation needed] light microscope for the 3D analysis of complete cells in the nanometer range.

A particularity of this technology compared with focusing techniques such as 4Pi microscopy, is the wide field exposures which allow entire cells to be depicted at the nano scale.

These are processes which modify the point spread function (PSF) of a microscope in a suitable manner to either increase the optical resolution, to maximize the precision of distance measurements of fluorescent objects that are small relative to the wavelength of the illuminating light, or to extract other structural parameters in the nanometer range.

[3][4] SMI can be combined with other super resolution technologies, for instance with 3D LIMON or LSI-TIRF as a total internal reflection interferometer with laterally structured illumination.

This SMI technique allowed to acquire light-optical images of autofluorophore distributions in the sections from human eye tissue with previously unmatched optical resolution.

By applying suitable laser optical precision processes, position and distances significantly smaller than the half-width of the point spread function (conventionally 200–250 nm) can be measured with nanometer accuracy between targets with different spectral signatures.

[12] The GFP gene has been introduced and expressed in many procaryotic and eucaryotic cells and the Nobel Prize in Chemistry 2008 was awarded to Martin Chalfie, Osamu Shimomura, and Roger Y. Tsien for their discovery and development of the green fluorescent protein.

The finding that these standard fluorescent molecules can be used extends the applicability of the SPMD method to numerous research fields in biophysics, cell biology and medicine.

At present, SPDM achieves 16 frames/sec with an effective resolution of 10 nm in 2D (object plane); approximately 2000 such frames are combined with SMI data (ca.

Using suitable dyes, even higher effective optical 3D resolutions should be possible[13] By combining SPDMphymod with SMI (both invented in Christoph Cremer´s lab in 1996) a 3D dual colour reconstruction of the spatial arrangements of Her2/neu and Her3 clusters was achieved.

For example, biomolecular machines (BMM) are highly complex nanostructures consisting of several large molecules and which are responsible for basic functions in the body cells.

Examples of biomolecular machines are nucleosomes which enable the DNA, a two meter long carrier of genetic information, to fold in the body cells in a space of a few millionth of a millimeter in diameter only.