Eos kiss x5

All photographs of lesion sites were taken by single-lens reflex camera (EOS Kiss X5; Canon, Tokyo, Japan) equipped with an 18- to 55-mm macro lens (Canon) and macro ring light (MR-14 EX; Canon). The photographing conditions were as follows: the ISO film speed was AUTO, shutter speed was 1/125 s, and F-value was 8.0. Photos at visual inspection and the IOM were taken with a flash in a room with white light, and those for the AVM were taken without a flash and with the room light turned off.

Each insect and flower sample was briefly shaken in 5 mL of sterilized 0.1% Tween in 0.15 M NaCl22 (link), then microbes were detached by ultrasonic dispersion for 20 s at 50% of maximum power (UR-21P, TOMY, Tokyo, Japan). After the detachment, 1 mL of the solution was stained with 4′,6-deamidino-2-phynylindole (DAPI)23 , filtered on a polycarbonate filter (0.2 μm pores, ϕ25 mm, Millipore, Billerica, MA, USA) placed on a nitrocellulose filter (0.45 μm pores, ϕ25 mm, Millipore) mounted in a glass holder for the filtration (i.e., two membrane filters were used for one filtration). In addition, 4 mL of the solution was filtered for DNA extraction, and the filter membrane was stored at −20°C until further processing. For each sample, three microscope pictures of DAPI-positive cells with blue excitation (460–490 nm; U-MWB2, Olympus, Tokyo, Japan) were taken at 400× magnification using an epifluorescence microscope (BX60; Olympus) and an attached digital-camera (EOS Kiss X5; Canon, Tokyo, Japan) as described previously24 . The microscope pictures were then processed with an automated image processing program, the “EBImage” and “biOps” packages of the software R.

The structures of OM microfluidics were observed by a scanning electron microscope (JEOL JSM-7500F). Prior to SEM observation, a 5-nm-thick osmium layer was deposited on the sample by sputter coating (Osmium Plasma Coater OPC60A, Filgen). OM channel topography was characterized by both an atomic force microscope (NanoWizard III, JPK instruments) and a digital microscope (VHX-7000, Keyence). Macro lens photos were taken by a DSLR camera (EOS Kiss X5, EFS 60 mm macro lens, Canon). Microscope photos were taken by an upright optical microscope (Axioscope A1 MAT, Carl Zeiss). The DSLR camera and optical microscope were white balanced using an 18% neutral grey card prior to photo taking. The reflectance spectra of OM channels were measured by a spectrometer (MCPD-3700, Otsuka Electronics) with a 210–820 nm light source (MC-2530, Otsuka Electronics, Japan). All reflectance spectra were normalized with the substrate (e.g., bare silicon wafer) as the reference. Film thickness was determined by the optical analysis software of the same manufacturer using Cauchy equation³⁸ . Contact angles were measured by an optical goniometric instrument (DSA25S, KRÜSS, GmbH) using the sessile drop technique.