ABSTRACT
Holographic microscopy has emerged as a tool for in situ imaging of microscopic organisms and other particles in the marine environment: appealing because of the relatively larger sampling volume and simpler optical configuration compared to other imaging systems. However, its quantitative capabilities have so far remained uncertain, in part because hologram reconstruction and image recognition have required manual operation. Here, we assess the quantitative skill of our automated hologram processing pipeline (CCV Pipeline), to evaluate the size and concentration measurements of environmental and cultured assemblages of marine plankton particles, and microspheres. Over 1 million particles, ranging from 10 to 200 µm in equivalent spherical diameter, imaged by the 4-Deep HoloSea digital inline holographic microscope (DIHM) are analyzed. These measurements were collected in parallel with a FlowCam (FC), Imaging FlowCytobot (IFCB), and manual microscope identification. Once corrections for particle location and nonuniform illumination were developed and applied, the DIHM showed an underestimate in ESD of about 3% to 10%, but successfully reproduced the size spectral slope from environmental samples, and the size distribution of cultures (Dunaliella tertiolecta, Heterosigma akashiwo, and Prorocentrum micans) and microspheres. DIHM concentrations (order 1 to 1000 particles ml-1) showed a linear agreement (r 2 = 0.73) with the other instruments, but individual comparisons at times had large uncertainty. Overall, we found the DIHM and the CCV Pipeline required extensive manual correction, but once corrected, provided concentration and size estimates comparable to the other imaging systems assessed in this study. Holographic cameras are mechanically simple, autonomous, can operate at very high pressures, and provide a larger sampling volume than comparable lens-based tools. Thus, we anticipate that these characterization efforts will be rewarded with novel discovery in new oceanic environments.
ABSTRACT
Bioabsorbable plating systems play an integral role in cranial vault remodeling. After experiencing a case of plate failure requiring emergent reexploration, we investigated the potential causes. We hypothesize that extended submersion in the molding bath during plate preparation might advance the rate of hydrolysis and compromise plate structural integrity. Using an absorbable poly-D/L-lactic acid plating system, we assessed the effect of extended submersion on plate strength and stiffness when loaded in a cantilever fashion and with pure tension. We assessed these differences with the Student t test and linear regression modeling. We also generated a computer model of the plates for finite element analysis. When left in the molding bath for extended periods, the plates changed color and lost strength. After 5 minutes, 30% of maximum plate load capacity was lost in a cantilever beam test (P < 0.001) consistent with use of a 15% thinner plate. Tensile testing revealed the initial elastic modulus of 6.42 +/- 0.13 GPa decreased 16% to 5.41 +/- 0.50 GPa after 5 minutes of submersion (P = 0.027). The changes in plate strength and elastic modulus both worsened with increased submersion times. Finite element analysis of the plates also predicted clinically significant increases in plate deviation under normal loading conditions. Our study demonstrates that extended submersion of absorbable plates during molding results in a significant loss of plate strength and stiffness. Further, our computer model predicts that these changes could result in an unacceptable plate deviation under normal loading conditions. Together, these data caution against overmolding of plates to avoid compromising their structural integrity.
