Insect Puparium Removal: A Method for Pupal Investigation

Abstract

Source: Fukutomi, Y., et al. Methods for Staging Pupal Periods and Measurement of Wing Pigmentation of Drosophila guttifera. J. Vis. Exp. (2018).

This article describes a technique to remove an insect’s puparium while keeping the inside pupa intact and therefore fit for further investigation. The example protocol shows the procedure with Drosophila guttifera; however, this technique is applicable to other Drosophila species.

Protocol

This protocol is an excerpt from Fukutomi et al., Methods for Staging Pupal Periods and Measurement of Wing Pigmentation of Drosophila guttifera, J. Vis. Exp. (2018).

1. Removing puparium

NOTE Pupae of Drosophila are covered by a structure called the puparium. An insect of Muscomorpha (flies) does not shed its larval cuticle at pupation; instead, it hardens the cuticle after apolysis, and uses it as a protective cover of the pupa, the puparium. A pupa residing inside a puparium has a true pupal cuticle, which is very soft and fragile....

Results

Figure 1. Illustration of removing puparium. (A) Place a pupa ventral side up on a piece of double-sided tape. Remove the anterior part of the puparium. (B) Break the puparium with forceps from the ventral .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Drosophila guttifera	The Drosophila Species Stock Center at the U.C. San Diego	15130-1971.10	Drosophila guttifera,  a fruit fly species used in this article
NICETACK double sided tape	Nichiban	NW-15SF	Double sided tape,   for removing puparium
Dumont #5 forceps	Fine Science Tools	11252-20	Forceps,  for removing puparium
Van Gogh VISUAL Paint brush	Talens Japan	GWVR-#5/0	Paint brush,  for removing puparium
Stereomicroscope	Olympus	SZX16	Stereomicroscope,  for morphological observation
Digital camera	Olympus	DSE-330-A	Digital camera,  for imaging
Falcon standard tissue culture dish	Corning	353002	Plastic Petri dish (diameter 60 mm x height 15 mm)