Study On The Relationship Between The AD-related Appsw Overexpressing Cell And DNA Folding Compression By STORM Technology

Alzheimer’s disease(AD)is a degenerative disease of the elderly.With the aging of society,ad has become a worldwide research hotspot and difficulty.The β-amyloid precursor protein(APP)gene,a mutation gene,is the earliest to be found in AD.Abnormal regulation(apparent)of APP results in a series of pathological changes.Overexpression of APP gene promotes neurons to enter the cell cycle and induces apoptosis,which leads to the occurrence and development of AD.The folding and compression of deoxyribonucleic acid(DNA)plays an important role in the regulation of gene expression.However,no evidence has been found in studies on DNA folding compression of Alzheimer’s disease.Existing optical microscopy techniques are limited by spatial resolution,it is difficult to study the pathogenesis of AD from the perspective of DNA folding compression.Stochastic optical reconstruction microscopy(STORM)is one of several super-resolution fluorescence microscopes that break through the diffraction of light in recent years.Its spatial resolution reaches 40 nm,which provides the possibility for us to study the relationship between gene and DNA folding compression and explore the pathogenesis of AD.In this paper,the experimental methods and researches were carried out around the folding and compression of DNA.The main research contents are as follows.1.The APPsw-overexpressing cells and the non-overexpressing cell cycle were synchronized by thymidine double-blocking method,and the release cycle analysis was carried out to analyze the gene replication transcription level and determine the degree of DNA folding and compression.2.Built STORM super-resolution imaging system,optimized imaging and calibrate resolution,and described STORM imaging simulation and algorithm.3.For the nano-scale imaging of DNA folding compression,we optimized the experimental conditions,determined the best experimental conditions such as dye,imaging buffer,imaging exposure time and laser power;after drift correction,nanoscale superresolution imaging of DNA folding and compression was realized.4.Imaging studies were performed on the DNA folding compression of APPsw overexpressing cells,and the differences were discussed in combination with flow cytometry analysis.Thedegree of DNA folding and compression is consistent with the results of flow cytometry.The change of DNA folding and compression results in overexpression of APP gene,which may be related to the occurrence and development of AD.In this study,DNA nano-scale super-resolution imaging was realized,and their nano-scale structural features were obtained.The differential characteristics of DNA folding compression of APPsw over-expressing cells were analyzed,which was consistent with the results of flow cytometry experiments.New clues are of great value for future research on early diagnosis and gene intervention therapy of AD.