Multifocal Optical Coherence Microscope

描述(申请人提供)：本项目的目的是开发一种多焦点光学相干显微镜(OCM)。建议的OCM将允许对未染色的生物样本进行非侵入性成像，具有毫秒的时间分辨率和三维的亚细胞空间分辨率。在这个项目中，我们希望实现以下三个具体目标：1)全视场OCM的建设：我们计划开发一款基于数码相机的全视场OCM。电光相位调制器将用于对参考光束进行快速、无振动的相位调制。将采用近红外光，以确保OCM成像的高穿透深度。四步相移算法将用于同时重建强度敏感和相敏的OCM图像。将增加偏振敏感成像能力，以实现对具有双折射特性的生物样品的高对比度成像。2)多焦OCM的研制：为了保证采样量的稳定性，我们将采用多焦点照明和虚拟针孔共焦成像策略来进一步改进全场OCM。虚拟针孔共焦成像和低相干选通机制的组合将提供一个两阶段的“过滤”系统，用于有效地抑制不需要的噪声光：1)计算机合成的虚拟针孔将被用来抑制焦外光；2)低相干选通机制将被用来抑制相邻采样体之间的串扰噪声光。使用带宽为100 nm的近红外光(中心波长：~800 nm)，我们预计深度分辨率优于3？m，横向(垂直于光轴)分辨率将受到光学衍射的限制。3)多焦OCM的功能测试：将使用一系列生物组织/细胞来评估所提出的OCM的强度敏感、相位敏感和偏振敏感的成像能力。还将对人体皮肤进行多模式OCM成像，初步测试该仪器的成像稳定性，用于活体应用。尽管拟议的OCM承诺为各种生物医学应用提供一个通用的成像平台，但该项目将重点放在成像系统的概念验证上。 公共卫生相关性：建议的多焦点光学相干显微镜可以提供快速、非侵入性的三维亚细胞空间分辨率成像。我们预计，多焦OCM将在细胞生物学、神经生物学、组织工程、皮肤科和眼科等领域获得广泛的应用。

DESCRIPTION (provided by applicant): The purpose of this project is to develop a multifocal optical coherence microscope (OCM). The proposed OCM will allow noninvasive imaging of unstained biological specimens, with millisecond temporal resolution and sub-cellular spatial resolution in three dimensions. During this project, we expect to achieve three specific aims as follows: 1) Construction of a full-field OCM: We plan to develop a digital camera based full-filed OCM. An electro- optic phase modulator will be used for rapid, vibration-free phase modulation of the reference beam. A near infrared light will be employed to ensure high penetrating depth of OCM imaging. A four-step phase shifting algorithm will be used for concurrent reconstruction of intensity-sensitive and phase-sensitive OCM images. Polarization-sensitive imaging capability will be added to achieve high contrast imaging of biological specimens with birefringence characteristics. 2) Development of a multifocal OCM: In order to ensure the stability of sampling volume, we will employ a multifocal illumination and virtual pinhole confocal imaging strategy for further improvement of the full-field OCM. The combination of the virtual pinhole confocal imaging and low coherence gating mechanism will provide a two-stage `filtering' system for effective rejection of undesired noise light: 1) Computer- synthesized virtual pinholes will be used to reject out-of-focus light; 2) Low coherence gating mechanism will be used to reject cross-talk noise light between adjacent sampling volumes. Using a near infrared light (center wavelength: ~800 nm) with 100 nm bandwidth, we anticipate a depth resolution better than 3 ¿m. The lateral (perpendicular to the light axis) resolution will be optical diffraction limited. 3) Functional test of the multifocal OCM: A series of biological tissues/cells will be used to assess the intensity-sensitive, phase-sensitive, and polarization-sensitive imaging capabilities of the proposed OCM. Multi-modal OCM imaging of human skin will be also implemented for preliminary test of imaging stability of the proposed instrument for in vivo application. Although the proposed OCM promises a versatile imaging platform for a variety of biomedical applications, this project will focus on proof-of-concept of the imaging system. PUBLIC HEALTH RELEVANCE: The proposed multifocal optical coherence microscope can provide fast, noninvasive imaging with sub-cellular spatial resolution in three dimensions. We anticipate that the multifocal OCM will find a wide range of applications in cell biology, neurobiology, tissue engineering, dermatology, and ophthalmology.