The analysis of transgenic calli, cells or plants were performed as previously described [62 (link)]. Fluorescence microscopy was carried out using the tdTomato filter set: 554-nm excitation and 581-nm emission wavelength with an Olympus stereo microscope model SZX12 (Olympus America, Center Valley, PA, USA) (for callus imaging) and an Olympus BX51 epifluorescence (for cell imaging). Confocal microscopy images were produced using a confocal Leica TCS SP8 microscope. The samples were excited with a 543 nm HeNe laser and fluorescence emission was collected from 590 to 610 nm for pporRFP. Fluorescence intensity was measured using a spectrofluorometry according to methods described by Millwood [69 (link)] with a Fluorolog®-3 system (Jobin-Yvon and Glen Spectra, Edison, NJ, USA). Triplicate spectra/peak emission absorbance was adjusted by removing the background signal from corresponding controls used for each sample. For each sample, the youngest fully expanded leaf from T0 lines was chosen to measure the intensity of fluorescence in non-transgenic control and putatively transgenic plants.
Bx51 epifluorescence
Manufactured by Olympus
Sourced in United States
The BX51 epifluorescence is a microscope system designed for fluorescence imaging. It provides high-resolution imaging capabilities for a wide range of fluorescent samples. The system features advanced optics and a precise control system to deliver consistent and reliable performance.
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Lab products found in correlation
2 protocols using bx51 epifluorescence
1
Fluorescence Microscopy Analysis of Transgenic Plants
2
In Vivo Rickettsial Adhesion Assay
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The quantitative mouse in vivo rickettsial adhesion assay was performed as described (1) . The principal concept of this anatomically-based inoculation model is that the more rickettsiae adhere to the luminal surfaces of the blood vessels, the fewer rickettsiae are detected in peripheral blood samples. Multiple visceral organs are fixed without perfusion rinse (to keep blood in the vessel lumens) for immunofluorescence (IF)-based histological studies to identify unattached rickettsiae, which are trapped in clots in vessel lumens (1) . Briefly, after an ordinarily lethal dose of R. australis (1 x 10 7 PFU/0.2µl; the LD50 is 1 x 10 6 PFU) was injected through the tail vein, rickettsial virulence (by plaque assay) was measured in a 1 µl blood sample collected from the orbital venous sinus (OVS) at different times until 1 hr post-infection (p.i.). Rickettsial antigens were detected with a rabbit polyclonal antibody against SFG rickettsiae (1:1000) overnight at 4°C. Nuclei were counter-stained with DAPI. Fluorescent images were analyzed using an Olympus BX51 epifluorescence or Olympus IX81 confocal microscope.
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