Study of ss-DNA Adsorption and Nano-mechanical Properties on Mica Substrate with Surface Forces Apparatus

Many DNA?based devices need to build stable and controllable DNA films on surfaces. However, the most com?monly used method of film characterization, namely, the probe?like microscopes which may destroy the sample and substrate. Surface Forces Apparatus(SFA) technique, specializing in surface interaction studies, is introduced to investigate the e ects of DNA concentration on the formation of single?stranded DNA(ss?DNA) film. The result demonstrates that 50 ng/μL is the lowest concentration that ss?DNA construct a dense layer on mica. Besides, it is also indicated that at di erent DNA concentrations, ss?DNA exhibit diverse morphology: lying flat on surface at 50 ng/μL while forming bilayer or cross?link at 100 ng/μL, and these ss?DNA structures are stable enough due to the repeatabil?ity even under the load of 15 mN/m. At the same time, an obvious adhesion force is measured:/m at 100 ng/μL, respectively, which is attributed to the ion?correlation e ect. M-6.5 mN/m at 50 ng/μL and-5.3 mNoreover, the atomic force microscopy(AFM) images reveal the entire surface is covered with wormlike ss?DNA and the measured surface roughness(1.8±0.2 nm) also matches well with the film thickness by SFA. The desorption behaviors of ss?DNA layer from mica surface occur by adding sodium salt into gap bu er, which is mainly ascribed to the decreased ion?ion cor?relation force. This paper employing SFA and AFM techniques to characterize the DNA film with flexibility and stable mechanical ability achieved by ion bridging method, is helpful to fabricate the DNA?based devices in nanoscale.

Noise exposure induced cochlear hair cell death pathways in guinea pig

Objective To understand the mechanism of noise exposure induced outer hair cells(OHCs) death pathways. Methods Thirty two guinea pigs were used in this study. The animals were either exposed for 4 h/day to broadband noise at 122 dB SPL (A-weighted) for 2 consecutive days or perfused with MNNG. After auditory test, the cochleae of animals were dissected. Propidium iodide (PI), a DNA intercalating fluorescent probe, was used to trace morphological changes in OHC nuclei. F-actin staining was used to determine missing OHCs. Caspase-3 was detected in living organ of Corti whole mounts using the fluorescent probe. The single strand DNA (ssDNA) in apoptotic OHCs in guinea pigs and apoptosis inducing factor (AIF) in hair cells in guinea pigs were examined by immunohistology method. Whole mounts of organ of Corti were prepared. Morphological and fluorescent changes were examined under a confocal microscope. Results (1) Both apoptotic and necrotic hair cells appeared following noise exposure. (2) Noise exposure induced single strand DNA in apoptotic OHCs but not in the normal OHCs. (3) Either after noise exposure or after MNNG perfusion, apoptotic OHCs were featured by nuclear condensation or fragmentation with caspase-3 activation, whereas necrotic OHCs were characterized by nuclear swelling without caspase-3 activation. (4) In normal organ of Corti, AIF was located in the mitochondria areas. After noise exposure, AIF was translocated from mitochondria in apoptotic and necrotic OHCs. Conclusion These findings indicate that noise exposure damages DNA in the OHC, which triggers action of Caspase-3. Subsequently, AIF is translocated to the nucleus, leading to DNA damage and OHCs death.