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网友留言-Dvanced microscopy tactics are described separately. Photos and colocalization profiles were-纽带成品网站超市-专业建站99元全包-送域名-免费招代理-纽带建站超市
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Dvanced microscopy tactics are described separately. Photos and colocalization profiles were
All measurements have been performed at room temperature. Sequences of pictures have been acquired consecutively till the number of events per frame was too low for SR imaging, thus a standard imaging experiment consisted of ca. 30,000 frames.Dvanced microscopy techniques are described separately. Photos and colocalization profiles had been analyzed applying ImageJ (NIH, Bethesda).GSDIM superresolution microscopyImage acquisition. Single molecule pictures were acquired on a home-built wide field microscope with EMCCD (IXON-DU-897; Andor Technology) detection. The samples have been continuously excited with 532nm laser diode (SDL-532-200T; Shanghai Dream Lasers Technology) and energy density was controlled with a neutral density filter in between 1kW/cm2 to 30kW/cm2. In order to achieve an optimal occasion quantity per frame, a 405nm PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21992055 laser (SDL405-LM-20-T; Shanghai Dream Lasers Technology) was employed to depopulate the dark states at a energy density of 0.6kW/cm2. This laser was switched on and off using a homemade pulse generation unit connected towards the fire signal of your EMCCD along with the blanking signal with the lasers. In this manner, the sample was irradiated for short periods, preferable between frames. Each laser beams were collinearly overlapped (DCLP 425 (DCXR) AHF, and focused (Achromat f200 lens) onto the back focal plane of a Leica Plan one hundred?objective, 1.25 NA, oil immersion, to achieve wide-field illumination. Single molecule fluorescence emission was collected by the identical objective, and separated from PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/23387799 the excitation beams by signifies of a dichroic mirror (Z488532rpc; Chroma Technologies). Emission filters have been selected for Alexa532 (562?0, Semrock Inc.) and Alexa568 (605?0, Chroma Technology) and placed in front on the detector. Excitation residual light was additional cleaned using a notch filter (NF01-532U-25; Semrock Inc.). The pixel size of 83?3nm2 around the focal plane was calibrated using a resolution test pattern (USAF 1951 RES TARGET 2IN, Edmund Optics). Normally, sequences of photos were recorded onto a 70?0 pixel region with the EMCCD camera (ca. six?m).Dvanced microscopy methods are described separately. Photos and colocalization profiles have been analyzed utilizing ImageJ (NIH, Bethesda).GSDIM superresolution microscopyImage acquisition. Single molecule photos were acquired on a home-built wide field microscope with EMCCD (IXON-DU-897; Andor Technologies) detection. The samples were constantly excited with 532nm laser diode (SDL-532-200T; Shanghai Dream Lasers Technology) and energy density was controlled using a neutral density filter amongst 1kW/cm2 to 30kW/cm2. So that you can obtain an optimal event quantity per frame, a 405nm PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/21992055 laser (SDL405-LM-20-T; Shanghai Dream Lasers Technologies) was used to depopulate the dark states at a power density of 0.6kW/cm2. This laser was switched on and off utilizing a homemade pulse generation unit connected towards the fire signal of your EMCCD and the blanking signal of the lasers. In this manner, the sample was irradiated for quick periods, preferable among frames. Sequences of photos had been acquired consecutively till the amount of events per frame was too low for SR imaging, therefore a Dimethocainemedchemexpress common imaging experiment consisted of ca.
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