Measurement of single cell refractive index, dry mass, volume, and density using a transillumination microscope.
Measurement of single cell refractive index, dry mass, volume, and density using a transillumination microscope.
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DOI：
10.1103/physrevlett.109.118105
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发表时间：
2012-09-14
影响因子：
8.6
通讯作者：
McCarty OJ
中科院分区：
文献类型：
--
作者：
Phillips KG;Jacques SL;McCarty OJ
相差显微镜在生物学领域已经变得无处不在，特别是在细胞形态的定性研究中。然而，定量相位恢复方法的使用及其与细胞折射率和干质量密度的联系仍然没有得到充分利用。这部分是由于相位和细胞质量测定对定制仪器的限制，涉及数学分析和禁止的样品扰动。我们介绍断层明场成像，一个访问的光学成像技术，使三维测量的细胞折射率和干质量密度使用标准的透照光学显微镜。该技术的有效性证明聚苯乙烯球。然后将该技术应用于测量红细胞的折射率、干质量、体积和密度。这种光学技术使一个简单而强大的手段进行定量调查的工程和生物标本在三维使用标准光学显微镜。
Phase contrast microscopy has become ubiquitous in the field of biology, particularly in qualitative investigations of cellular morphology. However, the use of quantitative phase retrieval methods and their connection to cellular refractive index and dry mass density remain under utilized. This is due in part to the restriction of phase and cellular mass determination to custom built instruments, involved mathematical analysis, and prohibitive sample perturbations. We introduce tomographic bright field imaging, an accessible optical imaging technique enabling the three dimensional measurement of cellular refractive index and dry mass density using a standard transillumination optical microscope. The validity of the technique is demonstrated on polystyrene spheres. The technique is then applied to the measurement of the refractive index, dry mass, volume, and density of red blood cells. This optical technique enables a simple and robust means to perform quantitative investigations of engineered and biological specimens in three dimensions using standard optical microscopes.