Cuticle Disruption: A Method to Collect Hemolymph from Drosophila Larvae

Published: April 30, 2023

Abstract

Source: Reimels, T. A. and Pfleger, C. M. Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae. J. Vis. Exp. (2016).

The Drosophila hemolymph serves as both blood and interstitial fluid, circulating throughout the body during the larva and adult stages. This video describes a protocol to collect larval hemolymph in order to quantify the circulating cells.

Protocol

This protocol is an excerpt from Reimels and Pfleger, Methods to Examine the Lymph Gland and Hemocytes in Drosophila Larvae, J. Vis. Exp. (2016). 1. Circulating Hemocyte Concentration To obtain larvae of roughly the same developmental stage for this assay, restrict egg collection by allowing females to lay eggs for a fixed time period of 2 – 6 hr. Collect larvae in dissecting dish wells filled with 1x phosphate buffered saline (P…

Materials

PBS tablets MP Biomedicals 2810305
dissecting dish Corning 7220-85
microcentrifuge tube Denville C2170
silicone dissecting pad, made from Sylgard 184 kit Krayden (distributed through Fisher) NC9644388 (Fisher catalog number) Made in petri dish by mixing components of Sylgard elastomer kit according to manufacturer instructions.
stereomicroscope Morrell Instruments (Nikon distributor) mna42000, mma36300 Nikon models SMZ1000 and SMZ645
tissue wipe VWR 82003-820
forceps Electron Microscopy Sciences 72700-DZ
p200 pipette Eppendorf 3120000054
Countess Automated Cell Counter Invitrogen C10227
Countess cell counting chamber slides Invitrogen C10283
hemocytometer Hausser Scientific 3200
trypan blue stain Life Technologies T10282

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Cite This Article
Cuticle Disruption: A Method to Collect Hemolymph from Drosophila Larvae. J. Vis. Exp. (Pending Publication), e20126, doi: (2023).

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