Department of Physics, University of Wisconsin, Milwaukee, WI 53201, USA.
Proc Natl Acad Sci U S A. 2010 Nov 23;107(47):20335-9. doi: 10.1073/pnas.1012432107. Epub 2010 Nov 8.
Achieving label-free, molecular-specific imaging with high spatial resolution in deep tissue is often considered the grand challenge of optical imaging. To accomplish this goal, significant optical scattering in tissues has to be overcome while achieving molecular specificity without resorting to extrinsic labeling. We demonstrate the feasibility of developing such an optical imaging modality by combining the molecularly specific stimulated Raman excitation with the photoacoustic detection. By employing two ultrashort excitation laser pulses, separated in frequency by the vibrational frequency of a targeted molecule, only the specific vibrational level of the target molecules in the illuminated tissue volume is excited. This targeted optical absorption generates ultrasonic waves (referred to as stimulated Raman photoacoustic waves) which are detected using a traditional ultrasonic transducer to form an image following the design of the established photoacoustic microscopy.
实现在深层组织中进行无标记、分子特异性、高空间分辨率成像通常被认为是光学成象的重大挑战。为了实现这一目标,必须克服组织中的显著光散射,同时在不依赖于外源标记的情况下实现分子特异性。我们通过将分子特异性受激拉曼激发与光声检测相结合,展示了开发这种光学成象模式的可行性。通过使用两个在频率上通过目标分子的振动频率分离的超短激发激光脉冲,只有被照亮的组织体积中的目标分子的特定振动能级被激发。这种靶向光吸收产生超声波(称为受激拉曼光声波),然后使用传统超声换能器进行检测,根据已建立的光声显微镜的设计形成图像。