Ae31 inverted epi fluorescence microscope

Bright-field and fluorescent images were captured using an AE31 inverted Epi-fluorescence microscope (Motic, Hong Kong, China) equipped with an IS1000 eyepiece (Tucsen, Fujian, China)^{17 (link)}. Excitation filter modules of green and blue fluorescent protein (GFP, BFP) channels were set at 480/30 and 350/50 nm, respectively. Emission filter modules of GFP and BFP channels were set at 535/40 and 460/50 nm, respectively.

Fluorescent and bright-field images were recorded using an AE31 inverted Epi-fluorescence microscope (Motic, Causeway Bay, Hong Kong) with an IS1000 eyepiece (Tucsen, Fujian, China) or an Eclipse 80i Epi-fluorescence microscope (Nikon, Tokyo, Japan) with the NIS-elements documentation software (Nikon). For the AE31 fluorescent microscope, excitation filters were set at 480/30, 350/50, and 560/40 nm for GFP, blue (BFP), and red fluorescent protein (RFP) channels, respectively. Emission filters were set at 535/40, 460/50, and 630/60 nm for GFP, BFP, and RFP channels, respectively. For the Eclipse 80i Epi-fluorescent microscope, excitation filters were set at 450–490, 340–380, and 510–560 nm for GFP, BFP, and RFP channels, respectively. Emission filters were set at 520, 435–485, and 590 nm for GFP, BFP, and RFP channels, respectively. Bright-field images were used to observe cell morphology. Intensity of fluorescent images was quantified using the UN-SCAN-IT software (Silk Scientific, Orem, UT, USA).