High resolution optical projection tomographic microscopy for 3D tissue imaging

Qin Miao; Jon Hayenga; Michael G. Meyer; Thomas Neumann; Florence Patten; Alan C. Nelson; Eric J. Seibel

doi:10.1117/12.873485

11 February 2011 High resolution optical projection tomographic microscopy for 3D tissue imaging

Qin Miao, Jon Hayenga, Michael G. Meyer, Thomas Neumann, Florence Patten, Alan C. Nelson, Eric J. Seibel

Proceedings Volume 7904, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII; 79040L (2011) https://doi.org/10.1117/12.873485
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Optical projection tomography (OPT) requires a large depth of field (DOF) of a low numerical aperture (NA) lens resulting in low resolution. However, DOF of a high NA objective can be extended by scanning the focal plane through the sample. This extended DOF image is called pseudoprojection, which is used by optical projection tomographic microscope (OPTM) for tomographic reconstruction. The advantage of OPTM is the acquisition of relatively high resolution and large depth of field concurrently. This method requires the working distance of the lens to be larger than the size of the sample, so proper lens should be chosen for samples of different sizes. In this paper, we imaged hematoxylin stained muntjac cells inside capillary tube with two different sizes. Two objective lenses with different NA are used for these two tubes. Experimental results show that resolution improves over 10 times in OPTM compared to conventional OPT, which make it possible for OPTM technique to resolve sub-cellular features for large samples. Therefore, OPTM can be used for 3D histological analysis of hematoxylin & eosin (H&E) stained biopsy specimen with sub-cellular resolution in the future.

Citation Download Citation

Qin Miao, Jon Hayenga, Michael G. Meyer, Thomas Neumann, Florence Patten, Alan C. Nelson, and Eric J. Seibel "High resolution optical projection tomographic microscopy for 3D tissue imaging", Proc. SPIE 7904, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XVIII, 79040L (11 February 2011); https://doi.org/10.1117/12.873485

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