Applicability, usability, and limitations of murine embryonic imaging with optical coherence tomography and optical projection tomography

Abstract

Abstract:We present an analysis of imaging murine embryos at various embryonic developmental stages (embryonic day 9.5, 11.5, and 13.5) by optical coherence tomography (OCT) and optical projection tomography (OPT). We demonstrate that while OCT was capable of rapid highresolution live 3D imaging, its limited penetration depth prevented visualization of deeper structures, particularly in later stage embryos. In contrast, OPT was able to image the whole embryos, but could not be used in vivo because the embryos must be fixed and cleared. Moreover, the fixation process significantly altered the embryo morphology, which was quantified by the volume of the eye-globes before and after fixation. All of these factors should be weighed when determining which imaging modality one should use to achieve particular goals of a study. References and links1. J. Beckers, W. Wurst, and M. H. de Angelis, "Towards better mouse models: enhanced genotypes, systemic phenotyping and envirotype modelling," Nat. Rev. Genet. 10(6), 371-380 (2009 440-443 (2000). 4. K. Paigen and J. T. Eppig, "A mouse phenome project," Mamm. Genome 11(9), 715-717 (2000). Proc. Natl. Acad. Sci. U.S.A. 91(9), 3530-3533 (1994). 9. M. Dhenain, S. W. Ruffins, and R. E. Jacobs, "Three-dimensional digital mouse atlas using high-resolution MRI," Dev. Biol. 232(2), 458-470 (2001 95-105 (1999). 33. U. Morgner, W. Drexler, F. X. Kärtner, X. D. Li, C. Pitris, E. P. Ippen, and J. G. Fujimoto, "Spectroscopic optical coherence tomography," Opt. Lett. 25(2), 111-113 (2000). 34. S. Rugonyi, C. Shaut, A. Liu, K. Thornburg, and R. K. Wang, "Changes in wall motion and blood flow in the outflow tract of chick embryonic hearts observed with optical coherence tomography after outflow tract banding and vitelline-vein ligation," Phys.

Optical coherence elastography of cold cataract in porcine lens

“… 35 To image deep inside the lens and visualize the formation of the cold cataract, we utilized a commercial swept source OCT (SS-OCT) system (OCS1310V2, Thorlabs Inc., New Jersey) with a central wavelength of 1300 nm, bandwidth of , and sweep rate of 200 kHz. 36 …”

Comparison and combination of rotational imaging optical coherence tomography and selective plane illumination microscopy for embryonic study

“…Nevertheless, for early stage embryos (e.g. E9.5), a depth of field of a few millimeters is sufficient for whole-body embryo imaging [12,39].…”

“…We have previously compared the performance of OCT and OPT for imaging murine embryos, where we observed similar results that OCT was unable to wholly image embryos beyond E9.5. OPT was able to image entire later stage embryos (up to E13.5) but requires a lengthy clearing and fixation procedure, while OCT can perform live imaging [39]. Although live OPT imaging of murine embryos has been demonstrated, only a limb bud was successfully imaged [42].…”

OptiJ: Open-source optical projection tomography of large organ samples

“…OPT can operate using either absorption/scattering of the sample (transmission OPT, tOPT) or fluorescence (emission OPT, eOPT) to generate image contrast. The use of OPT has been reported widely, and applications include the visualization of the 3D anatomy of mouse embryos (16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27), zebrafish (21,24,(28)(29)(30)(31)(32)(33)(34), drosophila (35)(36)(37)(38), plants (39,40), C.elegans (41), animal organs (22,27,(42)(43)(44) and other mesoscopic samples (45)(46)(47). Although major improvements in resolution (48,49), acquisition time (31), field of view (FOV) (21,40) and compatibility with other imaging techniques (22,28,50) have been made, most OPT applications require advanced technical expertise, expensive equipment, and bespoke software for reconstruction.…”