T 3400 sem

Adults were placed in 95% ethanol and cleaned in an ultrasonic cleaner (KQ118, Kunshan, China) for 10s, and rinsed with 95% ethanol several times. The heads were removed with dissecting needles under a stereomicroscope (Olympus SZX10, Tokyo, Japan) and then dehydrated in baths of 100% ethanol twice for 30 min each, before being transferred to a graded series of tert-butyl alcohol (TBA) solutions of 25%, 50%, and 75% (ethanol: TBA was 3:1; 1:1; 1:3), each for 15 min duration, and 100% TBA for 30 min duration. The samples were then placed into a freeze-drier (VFD-21S, SHINKKU VD, Japan) for 3h. The dried specimens were mounted on aluminum stubs using double-sided copper sticky tape and coated with gold/palladium (40/60) in a high-resolution sputter coater (MSP-1S, SHINKKU VD, Tokyo, Japan), and then examined with a T-3400 SEM (Hitachi, Tokyo, Japan) operated at 15 kV or Nova Nano SEM-450 (FEI, Hillsboro, OR, USA) at 5–10 kV. Fifteen individuals of each species were observed.

Adults were placed in 95% ethanol. Their heads were removed with dissecting needles under a stereomicroscope (Olympus SZX10, Tokyo, Japan) and then dehydrated in 100% ethanol two times for 30 min. transferred to a graded series of tert-butyl ethanol (TBA) solutions of 25%, 50%, and 75% (ethanol: TBA was 3:1; 1:1; 1:3) for 15 min at each concentration, and finally 100% TBA for 30 min. The samples were then placed into a freeze-drier (VFD-21S, SHINKKU VD, Tokyo, Japan) for 3h. The dried specimens were mounted on aluminum stubs using double-sided copper sticktape and coated with gold/palladium (40/60) in a high resolution sputter coater (MSP-1S, SHINKKU VD, Tokyo, Japan), and then observations were performed with a T-3400 SEM (Hitachi, Tokyo, Japan) operated at 15 kV or Nova Nano SEM-450 (FEI, Hillsboro, OR, USA) at 5–10 kV. Thirty individuals of this species were observed.