Phase Relief Imaging with Confocal Laser Scanning System

摘要

Confocal laser scanning microscopy (CLSM) has become one of the most important biomedical research tools today due to its noninvasive and 3-D abilities. It enables imaging in living tissue with better resolution and contrast, and plays a growing role among microscopic techniques utilized for investigating numerous biological problems. In some cases, the sample was phase-sensitive, thus we introduce a novel method named laser oblique scanning optical microscopy (LOSOM) which could obtain a relief image in transparent sample directly. Through the LOSOM system, mouse kidney and HeLa cells sample were imaged and 10x, 20x and 40x magnify objective imaging results were realized respectively. Also, we compared the variation of pinhole size versus imaging result. One major parameters of LOSOM is the distance between fluorescence medium and the sample. Previously, this distance was set to 1.2 mm, which is the thickness of the slide. The experiment result showed that decreasing d can increase the signal level for LOSOM phase-relief imaging. We have also demonstrated the application of LOSOM in absorption imaging modality, when the specimen is non-transparent.