A high-throughput method of hemolymph extraction from adult Drosophila without anesthesia

• Hemolymph collection from adult Drosophila is technically challenging.
• We obtain hemolymph samples from adult flies using a simple and inexpensive device.
• We detail how to construct and use this device.
• We demonstrate a use of this method by measuring hemolymph [Na+] and [K+].

A rapid and cost-effective method of sampling hemolymph from the model insect Drosophila melanogaster is needed for studies in several fields, including ionoregulatory physiology, metabolism, immunology and toxicology. Here, we describe the construction and use of a device that uses airflow and pressure to manipulate adult flies and extract high-volume hemolymph samples. This method is rapid and inexpensive, and does not require cold or CO2 anesthesia at any point in the sampling process, thus avoiding the possible confounding effects of these treatments on the biochemical properties of the hemolymph sampled. To demonstrate one use for this method, we measure active concentrations of Na+ and K+ in isolated hemolymph droplets from individual adult D. melanogaster using an ion-selective microelectrode technique.

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