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A Laser Capture Microdissection Protocol That Yields High Quality RNA from Fresh-frozen Mouse Bones
摘要： RNA yield and integrity are decisive for RNA analysis. However, it is often technically challenging to maintain RNA integrity throughout the entire laser capture microdissection (LCM) procedure. Since LCM studies work with low amounts of material, concerns about limited RNA yields are also important. Therefore, an LCM protocol was developed to obtain sufficient quantity of high-quality RNA for gene expression analysis in bone cells. The effect of staining protocol, thickness of cryosections, microdissected tissue quantity, RNA extraction kit, and LCM system used on RNA yield and integrity obtained from microdissected bone cells was evaluated. Eight-μm-thick frozen bone sections were made using an adhesive film and stained using a rapid protocol for a commercial LCM stain. The sample was sandwiched between a polyethylene terephthalate (PET) membrane and the adhesive film. An LCM system that uses gravity for sample collection and a column-based RNA extraction method were used to obtain high quality RNAs of sufficient yield. The current study focusses on mouse femur sections. However, the LCM protocol reported here can be used to study in situ gene expression in cells of any hard tissue in both physiological conditions and disease processes.
关键词： Issue 151,bones,gene expression,laser capture microdissection,mouse,RNA,RNA integrity number,Biology
更新于2025-09-12 10:27:22
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A Photonic System for Generating Unconditional Polarization-Entangled Photons Based on Multiple Quantum Interference
摘要： We present a high-performance source of unconditional polarization-entangled photons that have a high-emission rate, a broadband distribution, are degenerated and postselection free. The property of the source is based on the multiple quantum interference effect with a round-trip configuration of a Sagnac interferometer. The quantum interference effects make it possible to use the high generation efficiency of the polarization-entangled photons to process parametric down-conversion, and separate degenerated photon pairs into different optical modes without a postselection requirement. The principle of the optical system was described and experimentally used to measure the fidelity and Bell parameters, and also to characterize the generated polarization-entangled photons from a minimum of six combinations of polarization correlated data. The experimentally obtained fidelity and Bell parameters exceeded the classical local correlation limit and are clear evidence of the generation of unconditional polarization-entangled photons.
关键词： type-0,round-trip configuration,Sagnac interferometer,type-II,parametric down-conversion,Polarization-entangled photons,Engineering,Issue 151,quantum interference
更新于2025-09-11 14:15:04