Uplansapo 60 water immersion objective

Cells were imaged using an Olympus FV3000 inverted confocal microscope equipped with a UPLANSAPO ×60 water-immersion objective (NA = 1.20, Olympus). Bleaching of YFP was achieved with a 514 nm laser at 100% power with 10 iterations. CFP emission was recorded before and after bleaching of YFP. FRET efficiency was calculated as E = 100(A − B)/B, where B and A indicated the average intensity of the three images taken before and after bleaching, respectively. The background was subtracted from each image before the calculation^{18 (link)}.

Cells were observed using an inverted fluorescence microscope (IX83-ZDC, Olympus) fitted with a confocal unit (CSU-W1, Yokogawa), a cooled charge-coupled device (CCD) camera (ORCA-R2, Hamamatsu Photonics), a UPLANSAPO ×60 water-immersion objective (NA = 1.20, Olympus), and laser lines set at 458, 488, and 561 nm. Images were acquired using MetaMorph (Molecular Devices).
To analyze the structure of pits, intact roots were stained with safranin, and thereafter imaged using an Olympus FV3000 inverted confocal microscope equipped with a UPLANSAPO ×100 oil-immersion objective (NA = 1.40, Olympus). FV-OSR software (Olympus) was used to obtain high-resolution images.

An inverted fluorescence microscope (IX83-ZDC, Olympus, https://www.olympus-lifescience.com/) fitted with a confocal unit (CSU-W1, Yokogawa), a cooled CCD camera (ORCA-R2, Hamamatsu Photonics) or an EM-CCD camera (iXon3-888, ANDOR), an UplanSAPO 60× water-immersion objective (NA = 1.2, Olympus), an UplanSAPO 10× objective (NA = 0.4, Olympus), and laser lines set at 458, 488, and 561 nm was used. Images were acquired with MetaMorph software (Molecular Devices). An EM-CCD camera was used for recoding tagRFP-ROPGEF4^PRONE and GFP-ROPGAP3 in tobacco leaf epidermis.
To obtain omni-directional views of metaxylem vessels, an Olympus FV3000 inverted confocal microscope (Olympus) equipped with an UPLAN 60× water-immersion objective (NA = 1.2) and a laser line set to 514 nm was used.