6. Label-free selective plane illumination microscopy of tissue samples

Abstract

Listen

Translate article

Abstract type Basic Science Research. Presentation type Poster Presentation. Introduction There is an increasing demand for non-invasive, label-free, three-dimensional imaging techniques for characterisation of optically turbid samples such as tissue engineered constructs, used in Regenerative Medicine, and native tissue for medical diagnosis. Selective plane provide both high temporal and spatial information, deliver rapid results, are capable of assessing quality control criteria of engineered products and are applicable to a clinical environment. Many existing optical imaging techniques are not suitable for imaging samples that are non-transparent and turbid. In this project a unique technique which is Label-free Selective Plane Microscopy will be demonstrated. The imaging capabilities of this new method will be compared with existing techniques including bright field microscopy, fluorescence microscopy and confocal microscopy. Methodology Test samples of polystyrene microbeads were prepared to simulate tissue. Red (6 micron in diameter) and clear microbeads (8 micron in diameter) were used to represent tissues that are and are not highly perfused with blood. For comparison with fluorescence imaging techniques, samples containing fluorescent beads were also prepared. The samples were imaged using bright field microscopy, Label-free Selective Plane Microscopy, fluorescence microscopy and confocal microscopy. Native tissue samples were also studied and as above samples with different optical absorption and scattering properties. Chicken tendon and sheep kidney were used. Results Label-free Selective Plane Microscopy have the capability to image highly scattering and absorbing tissue in 3D. Label-free Selective Plane Microscopy (SPIM) is a fast method for imaging tissue samples with high lateral and axial resolution and good depth penetration. Conclusion Overall this method meets the demands of the current needs for 3D imaging tissue samples in a label-free manner. Label-free Selective Plane Microscopy directly provides excellent information about the structure of the tissue samples. This work has highlighted the superiority of Label-free Selective Plane Microscopy to current approaches to label-free 3D imaging of tissue.