Lasers and bubbles:Thermocavitation for needle-free jet injections

This thesis describes thermocavitation in microfluidic confinement, which is the generation of a vapor bubble by laser-heating of the liquid. During the growth of this bubble, it pushes the remaining liquid in the microfluidic channel out, creating a fast jet which can penetrate the skin. Therefore, this technique can be used to inject medication or other liquids, such as tattoo ink, without the use of a needle. Good control of the jet velocity requires understanding of the laser-liquid interaction, which is the focus of this thesis.The two most commonly used lasers, pulsed and continuous-wave (CW) lasers were compared. Both lasers were found to generate similar bubble dynamics, but the CW laser is slightly less efficient and reproducible. However, CW lasers are more affordable and smaller in size, which could increase their potential to be used in a commercial handheld device. To further investigate thermocavitation, numerical simulations of the laser heating were compared with experimental results of the CW-generated bubbles. These simulations suggest that nucleation always occurred when the maximum temperature was around 237 °C. Therefore, varying the laser beam size and power allows for control over the bubble dynamics.Absorption of the optical energy by a thin gold layer on the channel wall was investigated as an alternative to volumetric absorption by the liquid itself. Although this heating through the gold layer could generate bubbles and jets, it was less efficient and degradation of the layer occurred. Improvement through simulations and different choice of materials may overcome these problems.Finally, surface modifications were applied to the microfluidic channel in order to improve the jet dynamics. Alternating hydrophobic and hydrophilic stripes along the jet axis enable further control over the jet formation, through two influences: shaping the initial meniscus, as well as keeping the jet straight along the hydrophilic stripe. Overall, the results in this thesis contribute to the understanding and control of laser-generated bubbles in microfluidic confinement and the resulting jet. These results are vital for further development of laser-actuated jet injection.<br/

Laser beam properties and microfluidic confinement control thermocavitation

Thermocavitation, the creation of a vapor bubble by heating a liquid with a continuous-wave laser, has been studied for a wide range of applications. Examples include the development of an actuator for needle-free jet injectors, as the pumping mechanism in microfluidic channels and crystallization or nanoparticle synthesis. Optimal use in these applications require control over the dynamics of the laser-generated bubble through the laser power and beam radius. In contrast to pulsed lasers, for continuous-wave lasers the influence of the laser beam radius on the bubble characteristics is not fully understood. Here, we present a novel way to control the size of the beam from an optical fiber by changing the distance from the glass-liquid interface. We show that the increase in beam size results in a longer nucleation time. Numerical simulations of the experiment show that the maximum temperature at the moment of nucleation is 237$\pm$5{\deg}C and independent of laser parameters. Due to delayed nucleation for larger beam sizes, more energy is absorbed by the liquid at the nucleation instant. Consequently, a larger beam size results in a faster growing bubble, producing the same effect as reducing the laser power. We conclude that the total bubble energy only depends on the amount of absorbed optical energy and it is independent of the beam radius and laser power for any amount of absorbed energy. This effect contrasts with pulsed lasers, where an increase in beam radius results in a reduction of bubble energy. Our results are of relevance for the use of continuous-wave laser-actuated cavitation in needle-free jet injectors as well as other applications of thermocavitation in microfluidic confinement

Cavitation induced by pulsed and continuous-wave fiber lasers in confinement

Bubbles generated with lasers under confinement have been investigated for their potential use as the driving mechanism for liquid micro-jets in various microfluidic devices, such as needle free jet injectors. Here, we report on the study of bubble formation by a continuous-wave (CW) and a pulsed laser inside an open-ended microfluidic capillary. This results in a direct comparison between bubbles generated by laser sources emitting light in different time scales (ms and ns). The bubble kinetics represents an important parameter because it determines the available kinetic energy for a subsequent liquid jet. We show that the bubble growth rate increases linearly with the delivered energy for both the CW and the pulsed laser. Experiments show that at equal absorption coefficient, the bubble growth for both lasers is similar, which indicates that they can be used interchangeably for a jet generation. However, bubbles generated by a CW laser require more optical energy, which is due to heat dissipation. Furthermore, the bubbles generated by the CW laser show a slightly larger variation in size and growth rate for identical initial conditions, which we attribute to the stochastic nature of thermocavitation

Laser beam properties and microfluidic confinement control thermocavitation

Thermocavitation, the creation of a vapor bubble by heating a liquid with a continuous-wave laser, has been studied for a wide range of applications. Examples include the development of an actuator for needle-free jet injectors, as the pumping mechanism in microfluidic channels and nanoparticle synthesis. Optimal use in these applications requires control over the bubble dynamics through the laser power and beam radius. However, the influence of the laser beam radius on the bubble characteristics is not fully understood. Here, we present a way to control the beam radius from an optical fiber by changing the distance from the glass-liquid interface. We show that the increase in the beam size results in a longer nucleation time. Numerical simulations of the experiment show that the maximum temperature at nucleation is 237 ± 5 °C and independent of laser parameters. Delayed nucleation for larger beam sizes results in more absorbed energy by the liquid at the nucleation instant. Consequently, a larger beam size results in a faster growing bubble, producing the same effect as reducing the laser power. We conclude that the bubble energy only depends on the amount of absorbed optical energy and it is independent of the beam radius and laser power for any amount of absorbed energy. This effect contrasts with pulsed lasers, where an increase in the beam radius results in a reduction of bubble energy. Our results are of relevance for the use of continuous-wave laser-actuated cavitation in needle-free jet injectors as well as other applications of thermocavitation in microfluidic confinement.</p

Puzzling subunits of mitochondrial cytochrome reductase

The ubiquinol-cytochrome c reductase complex, like the other proton-pumping respiratory complexes of mitochondria, is an assembly of many different subunits. However, only a few of these subunits participate directly in the electron transfer and proton translocation. The roles of the other subunits are largely unknown. We discuss here some intriguing features of two of these subunits

Spherical probes for simultaneous measurement of rotational and translational diffusion in 3 dimensions

Real time visualization and tracking of colloidal particles with 3D resolution is essential for probing the local structure and dynamics in complex fluids. Although tracking translational motion of spherical colloids is well-known, accessing rotational dynamics of such particles remains a great challenge. Here, we report a novel approach of using fluorescently labeled raspberry-like colloids with an optical anisotropy to concurrently track translational and rotational dynamics in 3 dimensions. The raspberry-like particles are coated by a silica layer of adjustable thickness, which allows tuning the surface roughness. The synthesis and applicability of the proposed method is demonstrated by two types of probes: rough and smoothened. The accuracy of measuring Mean Squared (Angular) Displacements are also demonstrated by using these 2 probes dispersed in 2 different solvents. The presented 3D trackable colloids offer a high potential for wide range of applications and studies, such as probing crystallization dynamics, phase transitions and the effect of surface roughness on diffusion

The Acidic Tail of the Cdc34 Ubiquitin-conjugating Enzyme Functions in Both Binding to and Catalysis with Ubiquitin Ligase SCFC^(dc4*)

Ubiquitin ligases, together with their cognate ubiquitin-conjugating enzymes, are responsible for the ubiquitylation of proteins, a process that regulates a myriad of eukaryotic cellular functions. The first cullin-RING ligase discovered, yeast SCF^(Cdc4), functions with the conjugating enzyme Cdc34 to regulate the cell cycle. Cdc34 orthologs are notable for their highly acidic C-terminal extension. Here we confirm that the Cdc34 acidic C-terminal tail has a role in Cdc34 binding to SCF^(Cdc4) and makes a major contribution to the submicromolar K_m of Cdc34 for SCF^(Cdc4). Moreover, we demonstrate that a key functional property of the tail is its acidity. Our analysis also uncovers an unexpected new function for the acidic tail in promoting catalysis. We demonstrate that SCF is functional when Cdc34 is fused to the C terminus of Cul1 and that this fusion retains partial function even when the acidic tail has been deleted. The Cdc34-SCF fusion proteins that lack the acidic tail must interact in a fundamentally different manner than unfused SCF and wild type Cdc34, demonstrating that distinct mechanisms of E2 recruitment to E3, as is seen in nature, can sustain substrate ubiquitylation. Finally, a search of the yeast proteome uncovered scores of proteins containing highly acidic stretches of amino acids, hinting that electrostatic interactions may be a common mechanism for facilitating protein assembly

Identification of additional homologues of subunits VII and VIII of the ubiquinol-cytochrome <i>c</i> oxidoreductase enables definition of consensus sequences

AbstractThe Candida utilis QCR7 gene encoding subunit VII of the ubiquinol-cytochrome c oxidoreductase was isolated by functional complementation of the Saccharomyces cerevisiae subunit VII-null mutant. Several other subunit VII homologues as well as homologues for subunit VIII were identified by screening the GenBank database. Some of these homologues for subunit VII could only be identified as such using a consensus sequence that was derived from the multiple sequence alignment. Definition of the consensus should facilitate further analysis of structure/function relationships in this protein

Reproducibility of postural control measurement during unstable sitting in low back pain patients

<p>Abstract</p> <p>Background</p> <p>Postural control tests like standing and sitting stabilometry are widely used to evaluate neuromuscular control related to trunk balance in low back pain patients. Chronic low back pain patients have lesser postural control compared to healthy subjects. Few studies have assessed the reproducibility of the centre of pressure deviations and to our knowledge no studies have investigated the reproducibility of three-dimensional kinematics of postural control tests in a low back pain population. Therefore the aim of this study was to assess the test-retest reproducibility of a seated postural control test in low back pain patients.</p> <p>Methods</p> <p>Postural control in low back pain patients was registered by a three dimensional motion analysis system combined with a force plate. Sixteen chronic low back pain patients having complaints for at least six months, were included based on specific clinical criteria. Every subject performed 4 postural control tests. Every test was repeated 4 times and lasted 40 seconds. The force plate registered the deviations of the centre of pressure. A Vicon-612-datastation, equipped with 7 infra-red M1 camera's, was used to track 13 markers attached to the torso and pelvis in order to estimate their angular displacement in the 3 cardinal planes.</p> <p>Results</p> <p>All Intraclass Correlation Coefficients (ICC) calculated for the force plate variables did not exceed 0.73 (ranging between 0.11 and 0.73). As for the torso, ICC's of the mean flexion-extension and rotation angles ranged from 0.65 to 0.93 and of the mean lateral flexion angle from 0.50 to 0.67. For the pelvis the ICC of the mean flexion-extension angle varied between 0.66 and 0.83, the mean lateral flexion angle between 0.16 and 0.81 and the mean rotation angle between 0.40 and 0.62.</p> <p>Consecutive data suggest that the low test-retest reproducibility is probably due to a learning effect.</p> <p>Conclusion</p> <p>The test-retest reproducibility of these postural control tests in an unstable sitting position can globally be considered as rather moderate. In order to improve the test-retest reproducibility, a learning period may be advisable at the beginning of the test.</p

Computed tomographic analysis of the quality of trunk muscles in asymptomatic and symptomatic lumbar discectomy patients

Background: No consensus exists on how rehabilitation programs for lumbar discectomy patients with persistent complaints after surgery should be composed. A better understanding of normal and abnormal postoperative trunk muscle condition might help direct the treatment goals. Methods: A three-dimensional CT scan of the lumbar spine was obtained in 18 symptomatic and 18 asymptomatic patients who had undergone a lumbar discectomy 42 months to 83 months (median 63 months) previously. The psoas muscle (PS), the paraspinal muscle mass (PA) and the multifidus muscle (MF) were outlined at the L3, L4 and L5 level. Of these muscles, fat free Cross Sectional Area (CSA) and fat CSA were determined. CSA of the lumbar erector spinae (LES = longissimus thoracis + iliocostalis lumborum) was calculated by subtracting MF CSA from PA CSA. Mean muscle CSA of the left and right sides was calculated at each level. To normalize the data for interpersonal comparison, the mean CSA was divided by the CSA of the L3 vertebral body (mCSA = normalized fat-free muscle CSA; fCSA = normalized fat CSA). Differences in CSA between the pain group and the pain free group were examined using a General Linear Model (GLM). Three levels were examined to investigate the possible role of the level of operation. Results: In lumbar discectomy patients with pain, the mCSA of the MF was significantly smaller than in pain-free subjects (p = 0.009) independently of the level. The mCSA of the LES was significantly smaller in pain patients, but only on the L3 slice (p = 0.018). No significant difference in mCSA of the PS was found between pain patients and pain-free patients (p = 0.462). The fCSA of the MF (p = 0.186) and of the LES (p = 0.256) were not significantly different between both populations. However, the fCSA of the PS was significantly larger in pain patients than in pain-free patients. (p = 0.012). The level of operation was never a significant factor. Conclusions: CT comparison of MF, LES and PS muscle condition between lumbar discectomy patients without pain and patients with protracted postoperative pain showed a smaller fat-free muscle CSA of the MF at all levels examined, a smaller fat-free muscle CSA of the LES at the L3 level, and more fat in the PS in patients with pain. The level of operation was not found to be of importance. The present results suggest a general lumbar muscle dysfunction in the pain group, in particular of the deep stabilizing muscle system