Abstract
We report on the development of fluorescence Gabor domain optical coherence microscopy (Fluo GD-OCM), a combination of GD-OCM with laser scanning confocal fluorescence microscopy (LSCFM) for synchronous micro-structural and fluorescence imaging. The dynamic focusing capability of GD-OCM provided the adaptive illumination environment for both modalities without any mechanical movement. Using Fluo GD-OCM, we imaged ex vivo DsRed-expressing cells in the brain of a transgenic mouse, as well as Cy3-labeled ganglion cells and Cy3-labeled astrocytes from a mouse retina. The self-registration of images taken by the two different imaging modalities showed the potential for a correlative study of subjects and double identification of the target.
Highlights
Since its introduction in 1991 as a noninvasive biomedical imaging modality based on lowcoherence interferometry [1], optical coherence tomography (OCT) underwent numerous technical innovations aimed at improving the imaging performance in terms of sensitivity, speed, resolution and field of view
We report on the first combination of Gabor domain OCM (GD-OCM) [36] and laser scanning confocal fluorescence microscopy (LSCFM) with dual-source illumination, which we named Fluo GD-OCM
In this first integration of fluorescence imaging with GD-OCM, it is natural to plan custom optics at a later stage of the research, which will help resolve these limitations
Summary
Since its introduction in 1991 as a noninvasive biomedical imaging modality based on lowcoherence interferometry [1], optical coherence tomography (OCT) underwent numerous technical innovations aimed at improving the imaging performance in terms of sensitivity, speed, resolution and field of view. Approaches to merge the OCT and fluorescence optical systems using either a dual-cladding fiber in fiber-based interferometry [18,19,20,21,22] or a dichroic mirror in free-space interferometry [23,24] were proposed. These methods enabled synchronous - not parallel - dual imaging. Dual-source illumination has two benefits: firstly, various fluorophores, i.e., in the visible (VIS) spectrum, can be selected for fluorescence imaging, while the near-infrared (NIR) spectrum remains available for conventional OCT imaging; secondly, the optical signal of each imaging modality can be detected without any interference
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