Draw graphic suite 2017

The specimens were collected from Lake Issyk-Kul by hand-net and preserved in ethanol (80%). In the laboratory, the posterior half of the abdomen was excised with a dissection needle and placed in 10% KOH solution for twenty minutes; after maceration, the genitalia were cleaned in one drop of soapy water and transferred into a few drops of glycerin. The specimens were examined using a stereomicroscope (Leica Apo S8). The specimens were then stored in the Entomology Laboratory of the Department of Biology at the Sciences and Arts Faculty in Kastamonu University, Turkey. Illustrations of the new record were drawn by CorelDraw Graphic Suite 2017 (Corel, the UK).

Dorsal root ganglia and spinal cord sections were visualized using standard fluorescence microscopy (Olympus BX51) and confocal microscopy (Leica TCS SP8X, Wetzlar, Germany). For confocal microscopy, sequential scanning with a white light laser (70% power) tuned to excite AF488 (495 nm laser 47%), AF555 (532 nm 29% intensity), and AF647 (633 nm 53%) was used. Detectors were tuned to emission ranges of 504 to 542 nm for AF488, 550 to 597 nm for AF555, and 649 to 750 nm for AF647. Images (1024 × 1024 pixels) were taken with an oil immersion X20 or X40 objective with a frame average of 6. Spinal cord slices for CTB-labelled projections were optically sectioned (2-µm thick sections) and projected images were reconstructed (50 µm). Images were processed and analyzed using LAS Lite (Leica Microsystems, Wetzlar, Germany), Image J software (NIH), and CorelDraw 2017 (CorelDraw Graphic Suite 2017, Corel) software. Other than making moderate adjustments for contrast and brightness, the images were not manipulated in any way.