Development of a Microfluidic Device for CD4+ T Cell Isolation and Automated Enumeration from Whole Blood

The development of point-of-care, cost-effective, and easy-to-use assays for the accurate counting of CD4+ T cells remains an important focus for HIV-1 disease management. The CD4+ T cell count provides an indication regarding the overall success of HIV-1 treatments. The CD4+ T count information is equally important for both resource-constrained regions and areas with extensive resources. Hospitals and other allied facilities may be overwhelmed by epidemics or other disasters. An assay for a physician’s office or other home-based setting is becoming increasingly popular. We have developed a technology for the rapid quantification of CD4+ T cells. A double antibody selection process, utilizing anti-CD4 and anti-CD3 antibodies, is tested and provides a high specificity. The assay utilizes a microfluidic chip coated with the anti-CD3 antibody, having an improved antibody avidity. As a result of enhanced binding, a higher flow rate can be applied that enables an improved channel washing to reduce non-specific bindings. A wide-field optical imaging system is also developed that provides the rapid quantification of cells. The designed optical setup is portable and low-cost. An ImageJ-based program is developed for the automatic counting of CD4+ T cells. We have successfully isolated and counted CD4+ T cells with high specificity and efficiency greater than 90%.

Rapid quantification of rice (Oryza sativa) qualities based on adaptive near infrared spectroscopy

Determination of rice quality parameters is the key factor affecting sustainable agriculture practices. The main purpose of this present study is to develop prediction models based on adaptive near infrared spectroscopy (NIRS) for rapid quantification of rice qualities in form of protein content. Rice samples were obtained from several paddy field in Aceh province with different cultivars. Near infrared spectral data of rice samples were acquired and in wavelength range from 1000 to 2500 nm and recorded as diffuse reflectance spectrum. Prediction models were established using principal component analysis (PCA), principal component analysis (PCR) and partial least square regression (PLSR). The results showed that NIRS combined with PCA can classify rice samples based on their cultivars. Moreover, this approach with PCR and PLSR can also predicted and determined protein contents with satisfactory performance achieving maximum correlation coefficient (r) of 0.81 and ratio prediction to deviation (RPD) index of 2.84 for PCR and r of 0.90 and RPD of 3.19 for PLSR respectively. Based on achieved results, it may conclude that adaptive NIRS approach can be used to quantify rice qualities rapidly and non-destructively.

Rapid quantification of Psilocybin with reversed-phase HPLC and single-wavelength detection

The alkaloid psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine) and the neurologically active psilocin (4-hydroxy-N,N-dimethyltryptamine) are the foremost compounds of pharmaceutical interest in Psilocybe mushrooms. As these compounds are infrequently analyzed in analytical labs, validated methods for rapid purity analysis are lacking. Newfound therapeutic use has invigorated academic and commercial interests in the molecules and new methods of production and available products are expanding. As a result, high-throughput methods of analysis for psilocybin must be improved to promptly determine chemical differences between mushroom genera or other sources of psilocybin and psilocin, as well as refined product purity. To address this, we developed an inexpensive HPLC technique for the efficient quantification of psilocybin and psilocin by using readily available equipment and dilute reagents. Aqueous ammonium formate (0.143 mM) was found to be preferable over techniques with much higher buffer concentrations or stronger acids for controlling psilocybin Zwitterion resolution. The chromatographic run time satisfied high-throughput analytical requirements with an efficient total runtime under 2 minutes. A standard octadecyl silica (C18) column provided excellent resolution between psilocybin and psilocin signals. The quality of the method was validated using certified analytical reference standards and was found to be accurate (3.5% bias, Psilocybin), reliable (0.32% RSD), and efficient (Psilocybin k’ = 1.78).