Excelon evolve 512 emccd camera

Immediately after preparation, cells were imaged on a Zeiss Axio Observer Z1 inverted microscope with an EC Plan-Neofluar 100 $\times$ 1.45 N.A. oil-immersion objective. Images were collected with a Photometrics eXcelon Evolve 512 EMCCD camera controlled through Zeiss Zen software. Hoechst was imaged using 100 W halogen lamp excitation with a 365 nm excitation filter and 445/50 nm emission filter; CAL Fluor® Red 590 Dye was imaged using 561 nm laser excitation and a 600/37 nm bandpass emission filter. QD_13.3 and commercial QDs were imaged using 488 nm laser excitation and a 600/37 nm bandpass emission filter. QD_17.4 and QD_9.2 were imaged using 488 nm laser excitation and a 585 nm long-pass emission filter. Z-stack images of entire cells were collected in 0.22 μm increments. For each sample, 20 areas on the coverglass were selected at random for imaging. To obtain single-molecule fluorescence intensity values, dye-probes and QD-SAv conjugates dispersed in PBS were adsorbed on glass coverslips and imaged via epifluorescence microscopy using identical conditions to those used for FISH images. For each sample, videos during continuous excitation were acquired to identify single molecules by their distinct intensity time-traces using MATLAB algorithms^{14 (link)}.

Fluorescence microscopy of isolated QDs and cells was performed using wide-field illumination on a Zeiss Axio Observer Z1 inverted microscope with a ×100 1.45NA alpha Plan-Fluar oil immersion objective, 100 W halogen lamp illumination, 488 nm/100 mW OPSL laser, and 561 nm/40 mW diode laser units. Images were acquired using a Photometrics eXcelon Evolve 512 EMCCD camera through the Zeiss ZEN software. Excitation light was filtered using Semrock and Zeiss filters (G 365, BP 470 nm/40 nm, BP 482/18, BP 561/14 nm). Emission signals were filtered using Semrock bandpass filters (445/50, 525/50, 562/40, 600/37, and 732/68 nm). Brightfield images were acquired using transmitted-light illumination (12 V, 100 W Halogen lamp) with DIC prism III/0.55.