Fringe optimization for structured illumination super-resolution microscope with digital micromirror device

Structured illumination microscopy(SIM)is a promising super-resolution technique for imaging subcellular structures and dynamics due to its compatibility with most commonly usedffuorescent labeling methods.Structured illumination can be obtained by either laser interference or projection of fringe patterns.Here,we proposed a fringe projector composed of a compact multiwavelength LEDs module and a digital micromirror device(DMD)which can be directly attached to most commercial invertedffuorescent microscopes and update it into a SIM system.The effects of the period and duty cycle of fringe patterns on the modulation depth of the structured lightfield were studied.With the optimized fringe pattern,1:6×resolution improvement could be obtained with high-end oil objectives.Multicolor imaging and dynamics of subcellular organelles in live cells were also demonstrated.Our method provides a low-cost solution for SIM setup to expand its wide range of applications to most research labs in thefield of life science and medicine.

Characterization and functional analysis of chitinase family genes involved in nymph-adult transition of Sogatella furcifera

Chitinase degrades chitin in the old epidermis or peritrophic matrix of insects,which ensures normal development and metamorphosis.In our previous work,we comprehensively studied the function of SfCht7 in Sogatella furcifera.However,the number and function of chitinase genes in S.furcifera remain unknown.Here,we identified 12 full-length chitinase transcripts from S.furcifera,which included nine chitinase(Cht),two imaginal disc growth factor(IDGF),and one endo-β-N-acetylglucosaminidase(ENGase)genes.Expression analysis results revealed that the expression levels of eight genes(SfCht3,SfCht5,SfCht6-1,SfCht6-2,SfCht7,SfCht8,SfCht10,and SfIDGF2)with similar transcript levels peaked prior to molting of each nymph and were highly expressed in the integument.Based on RNA interference(RNAi),description of the functions of each chitinase gene indicated that the silencing of SfCht5,SfCht10,and SfIDGF2 led to molting defects and lethality.RNAi inhibited the expressions of SfCht5,SfCht7,SfCht10,and SfIDGF2,which led to downregulated expressions of chitin synthase 1(SfCHS1,SfCHS1a,and SfCHS1b)and four chitin deacetylase genes(SfCDA1,SfCDA2,SfCDA3,and SfCDA4),and caused a change in the expression level of two trehalase genes(TRE1 and TRE2).Furthermore,silencing of SfCht7 induced a significant decrease in the expression levels of three wing development-related genes(SfWG,SfDpp,and SfHh).In conclusion,SfCht5,SfCht7,SfCht10,and SfIDGF2 play vital roles in nymph–adult transition and are involved in the regulation of chitin metabolism,and SfCht7 is also involved in wing development;therefore,these genes are potential targets for control of S.furcifera.