Protein Microarray: "Theory" to "Real Practice"

Fueled by ever-growing genomic information and rapid developments of proteomics–the large scale analysis of proteins and mapping its functional role has become one of the most important disciplines for characterizing complex cell function. For building functional linkages between the biomolecules, and for providing insight into the mechanisms of biological processes, last decade witnessed the exploration of combinatorial and chip technology for the detection of bimolecules in a high throughput and spatially addressable fashion. Among the various techniques developed, the protein chip technology has been rapid. Recently we demonstrated a new platform called “Spacially addressable protein array” (SAPA) to profile the ligand receptor interactions. To optimize the platform, the present study investigated various parameters such as the surface chemistry and role of additives for achieving high density and high-throughput detection with minimal nonspecific protein adsorption. In summary the present poster will address some of the critical challenges in protein micro array technology and the process of fine tuning to achieve the optimum system for solving real biological problems.

UV Embossed Plastic Chip for Protein Separation and Identification

This report demonstrates a UV-embossed polymeric chip for protein separation and identification by Capillary Isoelectric Focusing (CIEF) and Matrix Assisted Laser Desportion/Ionization Mass Spectrometry (MALDI-MS). The polymeric chip has been fabricated by UV-embossing technique with high throughput; the issues in the fabrication have been addressed. In order to achieve high sensitivity of mass detection, five different types of UV curable polymer have been used as sample support to perform protein ionization in Mass Spectrometry (MS); the best results is compared to PMMA, which was the commonly used plastic chip for biomolecular separation. Experimental results show that signal from polyester is 12 times better than that of PMMA in terms of detection sensitivity. Finally, polyester chip is utilized to carry out CIEF to separate proteins, followed by MS identification.

Colliding and Moving Bose-Einstein Condensates: Studies of superfluidity and optical tweezers for condensate transport

In this thesis, two different sets of experiments are described. The first is an exploration of the microscopic superfluidity of dilute gaseous Bose- Einstein condensates. The second set of experiments were performed using transported condensates in a new BEC apparatus. Superfluidity was probed by moving impurities through a trapped condensate. The impurities were created using an optical Raman transition, which transferred a small fraction of the atoms into an untrapped hyperfine state. A dramatic reduction in the collisions between the moving impurities and the condensate was observed when the velocity of the impurities was close to the speed of sound of the condensate. This reduction was attributed to the superfluid properties of a BEC. In addition, we observed an increase in the collisional density as the number of impurity atoms increased. This enhancement is an indication of bosonic stimulation by the occupied final states. This stimulation was observed both at small and large velocities relative to the speed of sound. A theoretical calculation of the effect of finite temperature indicated that collision rate should be enhanced at small velocities due to thermal excitations. However, in the current experiments we were insensitive to this effect. Finally, the factor of two between the collisional rate between indistinguishable and distinguishable atoms was confirmed. A new BEC apparatus that can transport condensates using optical tweezers was constructed. Condensates containing 10-15 million sodium atoms were produced in 20 s using conventional BEC production techniques. These condensates were then transferred into an optical trap that was translated from the ‘production chamber’ into a separate vacuum chamber: the ‘science chamber’. Typically, we transferred 2-3 million condensed atoms in less than 2 s. This transport technique avoids optical and mechanical constrainsts of conventional condensate experiments and allows for the possibility of novel experiments. In the first experiments using transported BEC, we loaded condensed atoms from the optical tweezers into both macroscopic and miniaturized magnetic traps. Using microfabricated wires on a silicon chip, we observed excitation-less propagation of a BEC in a magnetic waveguide. The condensates fragmented when brought very close to the wire surface indicating that imperfections in the fabrication process might limit future experiments. Finally, we generated a continuous BEC source by periodically replenishing a condensate held in an optical reservoir trap using fresh condensates delivered using optical tweezers. More than a million condensed atoms were always present in the continuous source, raising the possibility of realizing a truly continuous atom lase.