Systematic Review and Meta-analysis of Methodological Quality in In Vivo Animal Studies of Subarachnoid Hemorrhage.

As a result of increased awareness of wide-spread methodological bias and obvious translational roadblocks in subarachnoid hemorrhage (SAH) research, various checklists and guidelines were developed over the past decades. This systematic review assesses the overall methodological quality of preclinical SAH research. An electronic search for preclinical studies on SAH revealed 3415 potential articles. Of these, 765 original research papers conducted in vivo in mice, rats, rabbits, cats, dogs, pigs, goats, and non-human primates with a focus on brain damage related to delayed cerebral vasospasm and early brain injury met the inclusion criteria. We found methodological shortcomings still to prevail in preclinical SAH research. In addition, basic animal characteristics were typically well described but important technical parameters of SAH induction were often underreported. None of the species, models, or techniques used in preclinical SAH research was methodologically superior to the others. Methodological quality of preclinical SAH research was independent of the number of citations or impact factor of a publication. Consequently, we suggest the SAH research community should consider strategies to improve preclinical research quality in their field, such as public platforms to (pre)register preclinical experiments, consequent support of open science policies, stricter editorial (and reviewer) control of (pre)existing guidelines, and increased efforts in education and training of good laboratory practice for the next generation of researchers.

Low-Cost 2D Index and Straightness Measurement System Based on a CMOS Image Sensor.

Accurate traceable measurement systems often use laser interferometers for position measurements in one or more dimensions. Since interferometers provide only incremental information, they are often combined with index sensors to provide a stable reference starting point. Straightness measurements are important for machine axis correction and for systems having several degrees of freedom. In this paper, we investigate the accuracy of an optical two-dimensional (2D) index sensor, which can also be used in a straightness measurement system, based on a fiber-coupled, collimated laser beam pointing onto an image sensor. Additionally, the sensor can directly determine a 2D position over a range of a few millimeters. The device is based on a simple and low-cost complementary metal-oxide-semiconductor (CMOS) image sensor chip and provides sub-micrometer accuracy. The system is an interesting alternative to standard techniques and can even be implemented on machines for real-time corrections. This paper presents the developed sensor properties for various applications and introduces a novel error separation method for straightness measurements.

Array based real-time measurement of fluid viscosities and mass-densities to monitor biological filament formation.

Liquid mass density and viscosity are fundamental characteristics of fluids. Their quantification by means of classical viscosity and density meters has several drawbacks: (i) the liquid-density and the viscosity cannot be measured simultaneously, (ii) sample volumes in the mL-range are consumed, (iii) the measurements cannot be multiplexed, and, (iv) the quantifications are time-consuming (minutes). Nano-mechanical transducers promise to overcome these limitations. We use fully clamped, gold coated silicon-nitride membranes with a thickness of 200 nm to measure liquid viscosity and density of samples of 1 µL volumes residing above the membrane in a miniature well. Photo-thermal actuation is used to excite the membrane, and an optical deflection system measures the response. From the response spectra, the eigenfrequency (f) and the quality (Q) factor are extracted and used to determine liquid density and viscosity by applying a three-point calibrated, simplified lumped model. We tested the system using calibrated solutions with viscosities in the range of 1-219 mPa s and mass densities between 998 kg m-3 and 1235 kg m-3. Real-time measurements were performed that characterize the polymerization of G-actin to F-actin filaments. The method presented promises to overcome the aforementioned limitations and thereby enables the real-time characterization of sub-µL sample volumes in a multiplexed manner.

Systematic Review of In Vivo Animal Models of Subarachnoid Hemorrhage: Species, Standard Parameters, and Outcomes.

In preclinical models, modification of experimental parameters associated with techniques of inducing subarachnoid hemorrhage (SAH) can greatly affect outcomes. To analyze how parameter choice affects the relevance and comparability of findings, we systematically reviewed 765 experimental studies of in vivo animal SAH models (2000-2014). During the last decade, we found marked increases in publications using smaller species and models for simulating acute events after SAH. Overall, the fewer types of species and models used did not correlate with an increased standardization in the experimental characteristics and procedures. However, by species, commonly applied, reliable parameters for each experimental SAH technique were identified in mouse, rat, rabbit, and dog models. Our findings can serve as a starting point for discussion toward a more uniform performance of SAH experiments, development of preclinical SAH common data elements, and establishment of standardized protocols for multicenter preclinical trials.

Real-time viscosity and mass density sensors requiring microliter sample volume based on nanomechanical resonators.

A microcantilever based method for fluid viscosity and mass density measurements with high temporal resolution and microliter sample consumption is presented. Nanomechanical cantilever vibration is driven by photothermal excitation and detected by an optical beam deflection system using two laser beams of different wavelengths. The theoretical framework relating cantilever response to the viscosity and mass density of the surrounding fluid was extended to consider higher flexural modes vibrating at high Reynolds numbers. The performance of the developed sensor and extended theory was validated over a viscosity range of 1-20 mPa·s and a corresponding mass density range of 998-1176 kg/m(3) using reference fluids. Separating sample plugs from the carrier fluid by a two-phase configuration in combination with a microfluidic flow cell, allowed samples of 5 µL to be sequentially measured under continuous flow, opening the method to fast and reliable screening applications. To demonstrate the study of dynamic processes, the viscosity and mass density changes occurring during the free radical polymerization of acrylamide were monitored and compared to published data. Shear-thinning was observed in the viscosity data at higher flexural modes, which vibrate at elevated frequencies. Rheokinetic models allowed the monomer-to-polymer conversion to be tracked in spite of the shear-thinning behavior, and could be applied to study the kinetics of unknown processes.

Single-cell lysis for visual analysis by electron microscopy.

The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.