Optical projection tomography (OPT) is the direct optical equivalent of X-ray computed tomography (CT). To obtain a larger depth of field, traditional OPT usually decreases the numerical aperture (NA) of the objective lens to decrease the resolution of the image. So, there is a trade-off between sample size and resolution. Commercial microfluidic systems can observe a sample in flow mode. In this paper, an OPT instrument is constructed to observe samples. The OPT instrument is combined with commercial microfluidic systems to obtain a three-dimensional and time (3D + T)/four-dimensional (4D) video of the sample. “Focal plane scanning” is also used to increase the images’ depth of field. A series of two-dimensional (2D) images in different focal planes was observed and compared with images simulated using our program. Our work dynamically monitors 3D OPT images. Commercial microfluidic systems simulate blood flow, which has potential application in blood monitoring and intelligent drug delivery platforms. We design an OPT adaptor to perform OPT on a commercial wide-field inverted microscope (Olympusix81). Images in different focal planes are observed and analyzed. Using a commercial microfluidic system, a video is also acquired to record motion pictures of samples at different flow rates. To our knowledge, this is the first time an OPT setup has been combined with a microfluidic system.

Optical projection tomography (OPT) is an advanced three-dimensional (3D) imaging technology, which uses the filtered backprojection (FBP) algorithm to recover the 3D volume with sufficient number of projections. As to in vivo imaging, it is urgent to reduce the number of projections because the acquisition time could be minimized. However, reconstructing from undersampled OPT data can lead to artifacts and the decline in image quality. The simultaneous iterative reconstruction technique (SIRT) is introduced to remove artifacts and improve the image quality. The image qualities reconstructed from FBP and SIRT separately are compared, and the structural similarity and peak signal-to-noise ratio are calculated. Through simulated phantoms and OPT data of in vivo zebrafish embryo, SIRT consistently outperforms FBP in terms of reduced artifacts and enhanced image contrast especially when the projection numbers are reduced. SIRT method can provide high-quality reconstruction with 50 or fewer projections, thereby significantly reducing the minimum acquisition time and light dose while maintaining reconstruction quality. Through optimization and GPU acceleration, the SIRT algorithm can converge faster so as to reduce the image processing time. To our knowledge, this is the first time one SIRT algorithm is used for reconstruction of sparse OPT data. The experimental results show that SIRT algorithm outperform FBP especially when the number of projections is reduced. In addition, SIRT performs better in the preservation of vascular signal, which is significant for the monitoring of angiogenesis.