Qcapture pro

Twenty-four hours after RNA transfection, cells were trypsinized and plated on 6-well plates (1000 cells/well) and cultured of 2 weeks. Then the colonies were captured with the Olympus SZX12 and QCapture Pro software. Cell colonies >0.1 mm in diameter were counted under a microscope.

Measurements of continuous size traits (total of 20 traits) on the left and right sides of the body were taken using a QImaging Micropublisher 5.0 camera mounted on an Olympus SZX9 dissecting microscope, with the aid of the QCapture Pro software. All images were processed using ImageJ software. Points of measurement on the tegmina were defined after examining multiple wings and are based on consistent points of venation and folding, approximating the length and width of the tegmina. On the raptorial arm we measured the ventral length of the coxa, femur, tibia and tarsus. Additionally we measured the length of the pronotum, and the femur and tibia length of each of the left and right meta- and mesothoracic limbs (hind- and mid-legs) (Table 1). To determine if there was a difference in the degree of asymmetry in continuous traits between dextral and sinistral males, we measured the level and direction of asymmetry of each continuous trait within an individual by subtracting the right variable from the left variable. Data were analysed using SPSS 17.0. All continuous data were tested for normality and none deviated significantly from a normal distribution (Kolmogorov-Smirnov, all P > 0.05).

The skin samples taken at days 4 and 7 were cut in half. All samples were Bouin-fixed, paraffin-embedded, and cut in semi-serial 6 μm-thick sections. The slides were stained with hematoxylin-eosin (HE) and Sirius red [27 (link)]. The re-epithelialization (length and thickness) and number of metaphases were evaluated under an Olympus BX41 light microscope with a 3.2 Megapixel Olympus Q-Color-3 Imaging System coupled to an image capture system (Q-Capture Pro). Types I and III collagen fibers were quantified by the Picro-Sirius technique under an optical microscope coupled to a polarizer (Attachment Nszh-KPO). All slides were analyzed using Image Pro-Plus (v. 4.5).