Stable Em-Rab7WT or Em-Rab7L8A NRK cells were imaged live at 37°C in complete phenol red-free DMEM using the Nikon Ti2-E inverted microscope with AX-R confocal microscopy system and Nikon elements software. To generate FRAP movies, images were collected for one minute prior to bleaching at 1 frame per 10 seconds using the AX-R resonant scanner. To bleach Em-Rab7+ endosomes/lysosomes, a 33.0625μm2 area was bleached for 10 seconds using the 488-laser line (laser power = 15) and the AX-R galvano scanner. Images were then collected post-bleach for 4 minutes, at 1 frame per 10 seconds, using the AX-R resonant scanner. All movies were processed identically following collection using Nikon Elements Denoising.ai and subsequent histogram matching bleach correction in FIJI.
Ax r confocal microscopy system
Manufactured by Nikon
The AX-R confocal microscopy system is a laboratory instrument designed for high-resolution imaging of microscopic samples. It utilizes a laser-based scanning system to capture detailed images of biological specimens, materials, and other micro-scale objects.
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Lab products found in correlation
2 protocols using ax r confocal microscopy system
1
Live Imaging of Rab7 Endosome/Lysosome Dynamics
2
Microscopic Imaging of Bacterial Conjugation
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A total of 5 µL of sample containing either Alexa-Fluor 568 labeled E. coli DH5α(F+) + pBAD donor cells mixed with the recipient E. coli dam-deficient seqA-YFP strain or pED208 ΔtraA donor cells mixed with recipient E. coli dam-deficient seqA-YFP strain or samples only containing E. coli DH5α(F+) harboring the pED208 + pBAD, pED208 ΔtraA, or E. coli dam-deficient seqA-YFP strain was applied to 25 × 75 mm 1.0-mm-thick glass microslide with or without an agar pad and covered with a 22 × 22 mm micro cover glass slide. The specimens were imaged using a Nikon Ti2-E inverted microscope with AX-R confocal microscopy system, 25 mm FOV resonant scanner, 25 mm FOV Galvano scanner, and an NSPARC detector unit. Images were captured using photomultipliers and the Nikon NIS Elements C software. Live cell imaging was performed at 23°C, and the samples were viewed using a 100×/NA1.52 oil objective with perfect focus during live image acquisition. Captured images and movies were processed using Elements Denoise.ai followed by compiling in FIJI software (29 (link)).
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