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Author Spotlight: A Precise and Quantifiable Method for Collecting Hemolymph from Small Arthropods
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A high quality method for hemolymph collection from honeybee larvae.

Nicole Pavan Butolo1, Patricia Azevedo2, Luciano Delmondes de Alencar2

  • 1Centro de Estudos de Insetos Sociais-CEIS, Instituto de Biociências-Programa de Pós Graduação em Biologia Celular e Molecular, Universidade Estadual Paulista 'Júlio de Mesquita Filho' (UNESP-SP), Rio Claro, SP, Brazil.

Plos One
|June 20, 2020
PubMed
Summary

A new method using ophthalmic scissors effectively extracts pure hemolymph from honeybee larvae. This technique improves bee immune system studies by providing uncontaminated samples for accurate hemocyte counting.

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Area of Science:

  • Apiculture and Insect Physiology
  • Environmental Toxicology

Background:

  • Bee decline is linked to pesticide exposure, impacting immune systems.
  • Counting hemocytes in hemolymph is crucial for assessing bee immune health.
  • Current hemolymph extraction from larvae is challenging and yields contaminated samples.

Purpose of the Study:

  • To develop a high-quality, reproducible method for honeybee larvae hemolymph extraction.
  • To compare the purity of hemolymph extracted using different tools.

Main Methods:

  • Tested extraction tools: ophthalmic scissors, 30G needle, forceps, syringe, capillaries.
  • Hemolymph extracted via incision (except scissors, from head region).
  • Assessed sample purity using turbidity analysis with a turbidimeter.

Main Results:

  • Ophthalmic scissors yielded the clearest, uncontaminated hemolymph samples.
  • Established a reference turbidity range (22,432.35–24,504.87 NTU) for high-quality samples.
  • Other tested methods resulted in contaminated samples.

Conclusions:

  • Ophthalmic scissors provide a superior method for honeybee larvae hemolymph extraction.
  • This technique enhances the reliability of immune response studies in bees.
  • Improved extraction methods are vital for understanding pesticide impacts on bee health.