Clearing and dissecting insects for internal skeletal morphological research with particular reference to bees

Author(s)

Diego Sasso Porto ^[1] ; Gabriel A.R. Melo ^[2] ; Eduardo A.B. Almeida ^[3]

Affiliation

^[1] Universidade de São Paulo. Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto. Departamento de Biologia - Brasil

^[2] Universidade Federal do Paraná. Departamento de Zoologia. Laboratório de Biologia Comparada de Hymenoptera - Brasil

^[3] Universidade de São Paulo. Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto. Departamento de Biologia - Brasil

• Document type: Journal article
• Source: Revista Brasileira de Entomologia; v. 60, n. 1, p. 109-113, 2016-03-00.

Abstract

ABSTRACT A detailed protocol for chemical clearing of bee specimens is presented. Dry specimens as well as those preserved in liquid media can be cleared using this protocol. The procedure consists of a combined use of alkaline solution (KOH or NaOH) and hydrogen peroxide (H2O2), followed by the boiling of the cleared specimens in 60–70% EtOH. Clearing is particularly useful for internal skeletal morphological research. This procedure allows for efficient study of internal projections of the exoskeleton (e.g., apodemes, furcae, phragmata, tentoria, internal ridges and sulci), but this process makes external features of the integument, as some sutures and sulci, readily available for observation as well. Upon completion of the chemical clearing process the specimens can be stored in glycerin. This procedure was developed and evaluated for the preparation of bees and other Apoidea, but modifications for use with other insect taxa should be straightforward after some experimentation on variations of timing of steps, concentration of solutions, temperatures, and the necessity of a given step. Comments on the long-term storage, morphological examination, and photodocumentation of cleared specimens are also provided. (AU)

• FAPESP's process: 12/22261-8 - Phylogenetic relationships of corbiculate bees (Hymenoptera: Apidae: Apini)
• Grantee: Diego Sasso Porto
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 11/09477-9 - Taxonomy, phylogeny, biogeography, and evolution of Neopasiphaeinae bees (Hymenoptera: Colletidae) using molecular and morphological data
• Grantee: Eduardo Andrade Botelho de Almeida
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 14/10090-0 - Comparative morphology of internal skeletal structures of the head of corbiculate bees (Apidae: Apini)
• Grantee: Diego Sasso Porto
• Support Opportunities: Scholarships abroad - Research Internship - Master's degree