Micro axial tomography: A miniaturized, versatile stage device to overcome resolution anisotropy in fluorescence light microscopy
| Authors | |
|---|---|
| Year of publication | 2011 |
| Type | Article in Periodical |
| Magazine / Source | Rev. Sci. Instrum. |
| MU Faculty or unit | |
| Citation | |
| Field | Use of computers, robotics and its application |
| Keywords | fluorescence; glass fibres; image reconstruction; image resolution; optical microscopes; optical microscopy; optical tomography; stepping motors |
| Description | To overcome anisotropic optical resolution or point localization precision in 3D fluorescence microscopy, micro axial tomography was suggested which allows object tilting on the microscopic stage. Here, we present a miniaturized device which can be implemented in a motor driven microscope stage. The footprint of this device corresponds to a standard microscope slide. A special glass fiber can manually be adjusted in the object space of the microscope lens. A stepwise fiber rotation can be controlled by a miniaturized stepping motor incorporated into the device. Test particles were fixed onto glass fibers, optically localized with high precision, and automatically rotated to obtain views from different perspective angles under which distances of corresponding pairs of objects were determined. From these angle dependent distance values, the real 3D distance was calculated with a precision in the ten nanometer range using standard microscopic equipment. Only very few images registered under different rotation angles are sufficient for full 3D reconstruction. |
| Related projects: |