Balancing the Initiation and Molecular Recognition Capabilities of Eosin Macroinitiators of Polymerization-Based Signal Amplification Reactions

Coupling polymerization initiators to molecular recognition events provide the ability to amplify these events and detect them using the formation of a cross-linked polymer as an inexpensive readout that is visible to the unaided eye. The eosin-tertiary amine co-initiation system, activated by visible light, has proven utility in this context when an average of three eosin molecules are coupled to a protein detection reagent. The present work addresses the question of how detection sensitivity is impacted when the number of eosin molecules per binding event increases in the range of two to fifteen. Unlike in other initiation systems, a non-monotonic relationship is observed between the number of initiators per binding event and the observed detection sensitivity.Burroughs Wellcome Fund (Career Award at the Scientific Interface)Massachusetts Institute of Technology. James H. Ferry Fund for Innovation in Research Educatio

Evaluating the sensitivity of hybridization-based epigenotyping using a methyl binding domain protein

Hypermethylation of CpG islands in gene promoter regions has been shown to be a predictive biomarker for certain diseases. Most current methods for methylation profiling are not well-suited for clinical analysis. Here, we report the development of an inexpensive device and an epigenotyping assay with a format conducive to multiplexed analysis.David H. Koch Institute for Integrative Cancer Research at MIT (First-year Graduate Fellowship)National Science Foundation (U.S.). Graduate Research FellowshipBurroughs Wellcome Fund (Career Award at the Scientific Interface)National Institute of Environmental Health Sciences (Grant P30-ES002109)Massachusetts Institute of Technology. James H. Ferry Fund for Innovation in Research Educatio

Systematic Study of Fluorescein-Functionalized Macrophotoinitiators for Colorimetric Bioassays

We report a systematic investigation of a set of photoreducible macrophotoinitiators for use in polymerization-based signal amplification. To test the dependence of photopolymerization responses on the number of photoinitiators localized per molecular recognition event, we gradually increased the number of photoinitiator molecules coupled to a constant scaffold macromolecule from an average of 7 per polymer to an average of 168 per polymer. To evaluate the capacity of the macrophotoinitiators to detect molecular recognition, we coupled neutravidin to these molecules to recognize biotin-labeled DNA immobilized on biochip test surfaces. Fluorescein macroinitiators were found to be useful in detecting molecular recognition above a threshold number of initiators per polymer. Above this threshold, increasing the number of initiators per macroinitiator resulted in increased signal strength. These findings demonstrate the feasibility of increasing the number of photoreducible initiators per binding event beyond three, the number used in previous studies, that the initiation reaction remains limiting in the range we investigated, and that the number of initiators per binding event in this system has a clear impact on assay sensitivity and signal strength

Thermal Structural Transitions and Carbon Dioxide Adsorption Properties of Zeolitic Imidazolate Framework‑7 (ZIF-7)

As a subset of the metal–organic frameworks, zeolitic imidazolate frameworks (ZIFs) have potential use in practical separations as a result of flexible yet reliable control over their pore sizes along with their chemical and thermal stabilities. Among many ZIF materials, we explored the effect of thermal treatments on the ZIF-7 structure, known for its promising characteristics toward H₂ separations; the pore sizes of ZIF-7 (0.29 nm) are desirable for molecular sieving, favoring H₂ (0.289 nm) over CO₂ (0.33 nm). Although thermogravimetric analysis indicated that ZIF-7 is thermally stabile up to ∼400 °C, the structural transition of ZIF-7 to an intermediate phase (as indicated by X-ray analysis) was observed under air as guest molecules were removed. The transition was further continued at higher temperatures, eventually leading toward the zinc oxide phase. Three types of ZIF-7 with differing shapes and sizes (∼100 nm spherical, ∼400 nm rhombic-dodecahedral, and ∼1300 nm rod-shaped) were employed to elucidate (1) thermal structural transitions while considering kinetically relevant processes and (2) discrepancies in the N₂ physisorption and CO₂ adsorption isotherms. The largest rod-shaped ZIF-7 particles showed a delayed thermal structural transition toward the stable zinc oxide phase. The CO₂ adsorption behaviors of the three ZIF-7s, despite their identical crystal structures, suggested minute differences in the pore structures; in particular, the smaller spherical ZIF-7 particles provided reversible CO₂ adsorption isotherms at ∼30–75 °C, a typical temperature range of flue gases from coal-fired power plants, in contrast to the larger rhombic-dodecahedral and rod-shaped ZIF-7 particles, which exhibited hysteretic CO₂ adsorption/desorption behavior

Thermosensitive Structural Changes and Adsorption Properties of Zeolitic Imidazolate Framework‑8 (ZIF-8)

We compared four types of ZIF-8 with varying sizes and shapes to determine their thermal-structural stability and derive appropriate thermal activation conditions and correlation between structural characteristics and adsorption properties. Under air, the ZIF-8 phase for all the samples was converted completely into the zinc oxide phase above ∼300 °C, though thermalgravimetric analysis (TGA) indicated that the original structure was stable to ∼300–350 °C. Longer exposures (∼30 d) suggested that thermal activation at ∼200 °C was appropriate for the removal of guest and/or solvent molecules under air without structural damage. Despite no noticeable change in X-ray diffraction (XRD) patterns after activation at 250 °C under air, the resulting BET surface areas and CO₂ adsorption amounts (at 1 bar and 30 °C) of ZIF-8s were reduced to ∼44–54 and ∼72–87%, respectively, as compared to those of appropriately activated ZIF-8s. It appears that after the activation at 250 °C under air, some Zn and N atoms were dissociated and converted to ZnOH and NOH, respectively, causing the partial structural damage of ZIF-8s