Vb 7010

Oil-Red-O staining was conducted as described previously [62 (link)]; images of stained animals were acquired using a Zeiss Axiovert S100 microscope and a CCD camera (Keyence VB-7010). To quantify Oil-Red-O staining, we manually determined the region of interest, which included the most of the anterior body region extending from the posterior edge of the pharynx to the vulva (S4B Fig) in each animal, and then measured red, green, and blue (RGB) intensities by using Image J software. Body fat levels were calculated as ratios of red to green intensities after subtraction of intensities measured in background regions.

The sample solutions containing CRP at concentrations ranging from 0 to 50 μg mL⁻¹ were introduced into the immunoassay microdevice by capillary action, and both ends of the microchannel were sealed using a PDMS prepolymer to prevent drying. Subsequently, fluorescence measurements were carried out for the various CRP concentrations using a fluorescence microscope equipped with a CCD camera (Keyence, VB-7010, Osaka, Japan) after 1 h of incubation.

Detached leaves of a susceptible pea line, PI 250438, were biolistically co-inoculated with tungsten particles coated with 800 ng of pCl-P3ΔARFPs and 200 ng of either of the pE2113 and WClMV vectors, as described previously38 (link)65 (link). The inoculated leaves were kept in petri dishes with moistened filter paper. The GFP signal was monitored using an epifluorescence microscope (VB 7010; Keyence, Osaka, Japan).

The interface tissues were fixed with formalin, acetic acid:ethanol:water (90:5:5, v/v/v). Fixed samples were sliced into 80 – 100 micrometer-thick sections with the Vibratome (VIB-1500, Vibratome Co. Ltd., St. Louis, MO, USA). Histochemical staining of sections was performed using a 0.5% (w/v) solution of Toluindine Blue O (1B-481, Waldeck GmbH & Co., Munster, Germany) in distilled water. Stained slices were observed and photographs were taken by using the Biological Microscope BX51 (Olympus, Tokyo, Japan) with the CCD camera, VB-7010 (KEYENCE, Osaka, Japan).

In the majority of Myo31DF^K2 homozygous males, the genitalia rotate 360° counterclockwise, which is the left-right inversed direction of their wild-type counterpart^{12 (link),14 (link)}. However, the rest of the males had genitalia whose rotation stopped before it was completed. The deviation in the rotation angle of the male genitalia (angle deviation) was defined as previously described with slight modifications^{14 (link)}. The angle formed by the midline of the abdomen and the dorsoventral line that passes though the anus and penis (middle of the claspers) was measured with Image J (http://rsb.info.nih.gov/ij/) on a photograph of the genitalia of each male (VB-7010, Keyence) (Fig. 2a,b). Based on this angle, the angle deviations were classified into eight groups: 0° (Left 22°-Right 22°), Right 45° (Right 22°-Right 67°), Right 90° (Right 67°-Right 112°), Right 135° (Right 112°-Right 157°), 180° (Right 157°- Left 157°), Left 135° (Left 157°-Left 112°), Left 90° (Left 112°-Left 67°), and Left 45° (Left 67°-Left 22°) (Fig. 2c).