Stably transfected cell lines are widely used in drug discovery and biological research to produce recombinant proteins. Generation of these cell lines requires the isolation of multiple clones, using time-consuming dilution methods, to evaluate the expression levels of the gene of interest. A new and efficient method is described for the generation of monoclonal cell lines, without the need for dilution cloning. In this new method, arrays of patterned cell colonies and single cell transfection are employed to deliver a plasmid coding for a reporter gene and conferring resistance to an antibiotic. Using a nanofountain probe electroporation system, probe positioning is achieved through a micromanipulator with sub-micron resolution and resistance-based feedback control. The array of patterned cell colonies allows for rapid selection of numerous stably transfected clonal cell lines located on the same culture well, conferring a significant advantage over slower and labor-intensive traditional methods. In addition to plasmid integration, this methodology can be seamlessly combined with CRISPR/Cas9 gene editing, paving the way for advanced cell engineering.
High performance long-wavelength infrared nBn photodetectors based on InAs/InAs1−xSbx type-II superlattices on GaSb substrate have been demonstrated. The photodetector's 50% cut-off wavelength was ∼10 μm at 77 K. The photodetector with a 6 μm-thick absorption region exhibited a peak responsivity of 4.47 A/W at 7.9 μm, corresponding to a quantum efficiency of 54% at −90 mV bias voltage under front-side illumination and without any anti-reflection coating. With an R × A of 119 Ω·cm2 and a dark current density of 4.4 × 10−4 A/cm2 under −90 mV applied bias at 77 K, the photodetector exhibited a specific detectivity of 2.8 × 1011 cm. Hz/W.
The dominant dark current mechanisms are identified and suppressed to improve the performance of midwave infrared InAs/GaSb type-II superlattice photodiodes at high temperatures. The optimized heterojunction photodiode exhibits a quantum efficiency of 50% for 2 μm thick active region without any bias dependence. At 150 K, R0A of 5100 Ω cm2 and specific detectivity of 1.05×1012 cm Hz0.5/W are demonstrated for a 50% cutoff wavelength of 4.2μm. Assuming 300 K background temperature and 2π field of view, the performance of the detector is background limited up to 180 K, which is improved by 25 °C compared to the homojunction photodiode. Infrared imaging using f/2.3 optics and an integration time of 10.02 ms demonstrates a noise equivalent temperature difference of 11 mK at operating temperatures below 120 K.
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