We report on the design and characterization of a low-temperature external cavity diode laser (ECDL) system for broad wavelength tuning. The performance achieved with multiple diode models addresses the scarcity of commercial red laser diodes below 633 nm, which is a wavelength range relevant to spectroscopy of many molecules and ions. Using a combination of multiple-stage thermoelectric cooling and water cooling, the operating temperature of a laser diode is lowered to −64 • C, more than 85 • C below the ambient temperature. The laser system integrates temperature and diffraction grating feedback tunability for coarse and fine wavelength adjustments, respectively. For two different diode models, single-mode operation was achieved with 38 mW output power at 616.8 nm and 69 mW at 622.6 nm, more than 15 nm below their ambient temperature free-running wavelengths. The ECDL design can be used for diodes of any available wavelength, allowing individual diodes to be tuned continuously over tens of nanometers and extending the wavelength coverage of commercial laser diodes.
Gene expression is tightly regulated by RNA-binding proteins (RBPs) to facilitate cell survival, differentiation, and migration. Previous reports have shown the importance of the Insulin-like Growth Factor II mRNA-Binding Protein (IGF2BP1/IMP1/ZBP1) in regulating RNA fate, including localization, transport, and translation. Here, we generated and characterized a knockout mouse to study RBP regulation. We report that IGF2BP1 is essential for proper brain development and neonatal survival. Specifically, these mice display disorganization in the developing neocortex, and further investigation revealed a loss of cortical marginal cell density at E17.5. We also investigated migratory cell populations in the IGF2BP1 − / − mice, using BrdU labeling, and detected fewer mitotically active cells in the cortical plate. Since RNA localization is important for cellular migration and directionality, we investigated the regulation of β -actin messenger RNA (mRNA), a well-characterized target with established roles in cell motility and development. To aid in our understanding of RBP and target mRNA regulation, we generated mice with endogenously labeled β -actin mRNA (IGF2BP1 − / − ; β -actin-MS2 + / + ). Using endogenously labeled β -actin transcripts, we report IGF2BP1 − / − neurons have increased transcription rates and total β -actin protein content. In addition, we found decreased transport and anchoring in knockout neurons. Overall, we present an important model for understanding RBP regulation of target mRNA.
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