SMW

Single Molecule Workstation

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 Obiettivo del progetto (Objective)

'This proposal aims at the development and application of an innovative single molecule workstation, enabling advances in the research happening throughout Europe and the rest of the world around the investigation of living cells. By combining three most advanced microscopic techniques into a single workstation, we aim at reaching a new quality level in the study of the molecular biology of living cells. The proposed single molecule workstation will be composed of three key elements: (i) inverted light microscope (ILM); (ii) atomic force microscope (AFM); and (iii) optical tweezers (OT). A true virtue of this initiative is that by combining several ultra-sensitive microscopy techniques into a single workstation completely new horizons for molecular biology related studies are opened. The aim of this combined ILM-AFM-OT setup is to look at the surface topography using high-resolution AFM, to study the distribution of cellular molecules using high sensitive fluorescence and contrast enhanced light microscopy (ILM), and to measure molecular interaction forces with ultra-sensitive optical tweezers. As a complementary method, photo-thermal nano-spectroscopy (PTNS) will be used to investigate spectroscopic properties of cellular material with a spatial resolution down to sub-100 nm which will enable chemical analysis of sub-cellular components. The combined setup will provide a qualitatively new level in microscopic studies, giving unprecedented versatility in the detection and monitoring of cellular events with highest spatial and temporal resolution. The proposed single workstation will be used for the study of the correlation between structure and function of living cells with applications in immunology and cancer research.'

Introduzione (Teaser)

Over recent years a number of imaging techniques have been developed that enable the visualisation and study of cells. A combination of the three most advanced microscopic techniques into a single platform would facilitate the simultaneous study of the structure and function of living cells.

Descrizione progetto (Article)

Understanding how cells function requires not only the characterisation of the molecular constituents of the cell, but also their spatial and temporal interplay. To facilitate this light and atomic force microscopy (AFM) techniques work in a complementary manner to study living cells.

While light microscopy allows the visualisation of the interior of cells, AFM provides resolution at the nanometre scale facilitating the study of mechanical interactions of membrane components. Optical tweezers (OTs) further enable the observation of molecular interactions inside the cell.

The EU-funded ?Single molecule workstation? (SMW) project aimed to integrate all conceivable functions of these complementary technologies into a single and unified single molecule workstation (SMW) concept. The proposed instrumentation consisted of inverted light microscope (ILM), AFM and OT technologies.

By combining these ultra-sensitive microscopy techniques into a single platform, partners wished to open up completely new horizons for molecular biology-related studies. The setup allowed for simultaneous surface topography using high-resolution AFM, study of the distribution of cellular molecules with enhanced ILM, and measurement of molecular interaction forces with ultra-sensitive OT. As a complementary method, photo-thermal nano-spectroscopy (PTNS) was used to investigate spectroscopic properties of cellular material with a high spatial resolution, enabling chemical analysis of sub-cellular structures.

The workstation was designed to integrate AFM and ILM from a mechanical, optical and electronic perspective. Additionally, software for the combination of these technologies coupled with a user interface was developed. Project partners generated a prototype of a combined AFM-ILM instrument and tested its applicability for cancer detection and immunological T cell activation.

The optimised SMW platform offers the unique opportunity to simultaneously study the interior and exterior of cells with high resolution. Such a tool is expected to have a major impact on the biology community.