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4 March 2019 Pseudo-volumetric fluorescence endomicroscopy with a fiber bundle (Conference Presentation)
Jean-Marc Tsang, Jason Ritt, Jerome Mertz
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Proceedings Volume 10854, Endoscopic Microscopy XIV; 1085411 (2019) https://doi.org/10.1117/12.2510407
Event: SPIE BiOS, 2019, San Francisco, California, United States
Abstract
The development of high resolution, 3D imaging endomicroscopy has recently been gaining interest due to needs in in vivo studies of neuronal dynamics and interactions in deep brain tissue. However, most types of endomicroscopy techniques adopting micro-objectives can hardly perform 3D imaging due to the lack of minimally invasive scanning mechanism. Here, we present a GRIN-lens based fluorescence endomicroscopy capable of obtaining pseudo 3D image of a fluorescent sample without scanning the probe. We use a confocal laser scanning microscope (CLSM) to scan the proximal end of a coherent fiber bundle and obtain an extended depth of focus at the distal end. Preliminary results using fluorescent beads showed that a single bead can be detected throughout hundreds of microns depth. In the detection path, we use two reflecting pinholes of different size to form two confocal and one non-confocal images of the proximal end of the fiber bundle - one pinhole conjugates to the single core being illuminated and the other pinhole conjugates to the surrounding cores. The intensity ratio of an object in these three images almost explicitly depends on its distance from the distal end of the fiber bundle. Thus, we can determine the relative depths of objects in a volume sample in a single acquisition. Future works will include calibrating the setup to obtain the true depth instead of relative depth and applying this technique to brain samples.
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Jean-Marc Tsang, Jason Ritt, and Jerome Mertz "Pseudo-volumetric fluorescence endomicroscopy with a fiber bundle (Conference Presentation)", Proc. SPIE 10854, Endoscopic Microscopy XIV, 1085411 (4 March 2019); https://doi.org/10.1117/12.2510407
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KEYWORDS
Endomicroscopy
Luminescence
Brain
Confocal microscopy
Stereoscopy
3D image processing
Image resolution
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