Kam Z, Hanser B, Gustafsson M G, Agard D A, Sedat J W
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Proc Natl Acad Sci U S A. 2001 Mar 27;98(7):3790-5. doi: 10.1073/pnas.071275698.
Light microscopy of thick biological samples, such as tissues, is often limited by aberrations caused by refractive index variations within the sample itself. This problem is particularly severe for live imaging, a field of great current excitement due to the development of inherently fluorescent proteins. We describe a method of removing such aberrations computationally by mapping the refractive index of the sample using differential interference contrast microscopy, modeling the aberrations by ray tracing through this index map, and using space-variant deconvolution to remove aberrations. This approach will open possibilities to study weakly labeled molecules in difficult-to-image live specimens.
对诸如组织等厚生物样本进行光学显微镜观察时，往往会受到样本自身折射率变化所引起的像差限制。对于实时成像而言，这个问题尤为严重，由于固有荧光蛋白的发展，实时成像目前是一个备受关注的领域。我们描述了一种通过以下方式在计算上消除此类像差的方法：使用微分干涉对比显微镜对样本的折射率进行映射，通过该折射率图进行光线追踪来模拟像差，并使用空间可变去卷积来消除像差。这种方法将为研究难以成像的活样本中弱标记分子开辟可能性。
