Blood Cell Separation Using Polypropylene-Based Microfluidic Devices Based on Deterministic Lateral Displacement.

Mammalian blood cell separation methods contribute to improving the diagnosis and treatment of animal and human diseases. Microfluidic deterministic lateral displacement (DLD) devices can sort cells based on their particle diameter. We developed microfluidic DLD devices with poly(propylene)-based resin and used them to separate bovine and human red blood cells (RBCs) and white blood cells (WBCs) without electric devices. To determine the critical cut-off diameter (Dc) of these devices, we used immunobeads with a diameter of 1-20 µm. The Dc values of the microfluidic DLD devices for the immunobeads in the experiments were similar to the calculated Dc values (8-10 µm). Results from bovine blood cell separation experiments suggest that lymphocytes and neutrophils can be separated from diluted, whole blood. Human RBCs were occasionally observed in the left outlet where larger particles with diameters closer to the Dc value were collected. Based on the Dc values, human neutrophils were sorted to the left outlet, whereas lymphocytes were observed in both outlets. Although microfluidic channel optimization is required for the concentration of sorted cells, the microfluidic DLD device prepared with a poly(propylene)-based resin has the potential for clinical use.

Relationship between Porcine Sperm Motility and Sperm Enzymatic Activity using Paper-based Devices.

Mammalian sperm motility has traditionally been analyzed to determine fertility using computer-assisted semen analysis (CASA) systems. To develop low-cost and robust male fertility diagnostics, we created a paper-based MTT assay and used it to estimate motile sperm concentration. When porcine sperm motility was inhibited using sperm enzyme inhibitors for sperm enzymes related to mitochondrial activity and glycolysis, we simultaneously recorded sperm motility and enzymatic reactivity using a portable motility analysis system (iSperm) and a paper-based MTT assay, respectively. When using our paper-based MTT-assay, we calculated the area mean value signal intensity (AMV) to evaluate enzymatic reactivity. Both sperm motility and AMV decreased following treatment with iodoacetamide (IODO) and 3-bromopyruvic acid (3BP), both of which are inhibitors of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found a correlation between recorded motility using iSperm and AMV from our paper-based assay (P < 0.05), suggesting that a sperm-related enzymatic reaction is involved in sperm motility. Under this protocol, MTT reduction was coupled with catalysis of GAPDH and was promoted by electron transfer from NADH. Based on this inhibitor study, sperm motility can be estimated using our paper-based MTT-assay.

Preservation of the auditory nerve function after translabyrinthine removal of vestibular schwannoma.

OBJECTIVE: To evaluate the function of the postoperative auditory nerve preserved after translabyrinthine (TL) vestibular schwannoma (VS) removal. METHODS: Fifteen patients, who underwent unilateral VS resection via a TL approach, were preserved auditory nerve anatomically. The size and location of VS were measured on MRI preoperatively. After surgery, the electrical stimulation test (EST) or electrically evoked auditory brainstem response (EABR) was performed. RESULTS: Four cases (27%) out of fifteen patients were retained the functional integrity of the auditory nerve after surgery. The maximum tumor size in the group with a positive response to EST or EABR was significantly smaller than that in the group with a negative response to EST. It seems to be difficult to preserve auditory nerve function in cases where a tumor extends to the fundus of internal auditory canal. CONCLUSION: These results suggest that size of tumor and/or extension of tumor to the fundus might be important factors in preserving the auditory nerve function even if using a TL approach.