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, …
Representative 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 side. (C) After breaking the puparium, take out the pupa using a paintbrush.
Divulgations
The authors have nothing to disclose.
Materials
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)