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Protocol for Drosophila ovary dissection
<ol>
<li>Feed flies with yeast 1-2 days prior to dissecting them to fatten up the ovaries.</li>
<li>Anaesthetize flies using carbon dioxide stream.</li>
<li>Using a pair of tweezers, select a female fly.</li>
<li>Submerge the female fly into 1X PBS.</li>
<li>Grab the fly at its lower thorax with a pair of tweezers.</li>
<li>Tug gently at the lower abdomen with another pair of tweezers until the internal organs in the abdomen are exposed.</li>
<li>Look for the pair of ovaries and detach it from other organs (e.g. the intestines). </li>
<li>Tease apart the ovarioles (if ovaries are to be used for immunostaining or in situ hybridization). </li>
<li>Keep ovaries in ice-cold 1X PBS while dissecting the next fly.</li>
</ol>

<ol><li>Costa, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+Drosophila+fragile+X+protein+functions+as+a+negative+regulator+in+the+orb+autoregulatory+pathway%5BTitle%5D)+AND+%22Dev+Cell%22%5BJournal%5D)">The Drosophila fragile X protein functions as a negative regulator in the orb autoregulatory pathway.</a> Dev Cell. 8, 331-342 (2005).</li>
<li>Tan, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+autoregulatory+feedback+loop+directs+the+localized+expression+of+the+Drosophila+CPEB+protein+Orb+in+the+developing+oocyte%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">An autoregulatory feedback loop directs the localized expression of the Drosophila CPEB protein Orb in the developing oocyte.</a> Development. 128, 1159-1169 (2001).</li>
</ol>

Here we demonstrate a method of dissecting ovaries in Drosophila. As the method of culturing the different cell types that exist in the Drosophila ovary has yet been defined, this is a rapid method to obtain the Drosophila ovarian tissue. Essentially, we show that dissecting Drosophila ovaries involves a simple two- or three-step procedure. Subsequent treatment of these dissected ovaries depend on the downstream experiments that will be performed.

<table><tbody><tr><td>Forceps</td><td>Tool</td><td>Sigma-Aldrich</td><td>T43537</td><td></td></tr><tr><td>Glass dissecting dish</td><td>Tool</td><td>Pyrex</td><td>7220-85</td><td></td></tr></tbody></table>

dissection of drosophila ovaries

Dissection of Drosophila Ovaries

Research

JoVE Journal

Biology

In insects, the oenocytes produce cuticular hydrocarbon compounds. These compounds protect against desiccation and facilitate chemical communication.  Here we demonstrate a dissection technique used to isolate the oenocytes from adult Drosophila melanogaster, and illustrate how this preparation can be utilized to study genes involved in hydrocarbon synthesis.

Dissection of Oenocytes from Adult Drosophila melanogaster

How niches and stem cells form during development is an important question with practical implications. In the Drosophila ovary, germ line stem cells and their somatic niches form during larval development. This video demonstrates how to dissect, stain and mount female gonads from late third instar (LL3) Drosophila larvae.

Dissection and Staining of Drosophila Larval Ovaries

The limiting factor in the use of the adult Drosophila eye to study neurodegeneration and cell biology is the difficult imaging of intracellular processes. We describe the dissection of single ommatidia to generate a bona-fide primary neuronal cell culture, which can be subject to drug treatment and advanced imaging.

Single Drosophila Ommatidium Dissection and Imaging

The Drosophila egg chamber is an excellent model for studying the mechanisms of mRNA localization. In order to capture the dynamic events that underpin the processes of localization, rapid high resolution imaging of live tissue is required. Here, we present a protocol for dissection and imaging of live samples with minimal disruption.

Preparing Individual Drosophila Egg Chambers for Live Imaging

This video describes the general anatomy of the Drosophila ovary and features a method of dissecting ovaries for live or fixed imaging.

Drosophila melanogaster Ovary Dissection: A Technique for Ex Vivo Tissue Preparation

The upright imaging method described in this protocol allows for the detailed visualization of the poles of a developing Drosophila melanogaster egg.  This end-on view provides a new perspective into the arrangements and morphologies of multiple cell types in the follicular epithelium.

Upright Imaging of Drosophila Egg Chambers

Analysis of the mitochondrial structure-function relationship is required for a thorough understanding of the regulatory mechanisms of mitochondrial functionality. Specific methods for studying mitochondrial structure and function in live and fixed Drosophila ovaries are described and demonstrated in this paper.

Studying Mitochondrial Structure and Function in Drosophila Ovaries

We describe a protocol for dissection, fixation, and immunostaining of steroidogenic organs in Drosophila larvae and adult females to study steroid hormone biosynthesis and its regulatory mechanism. In addition to steroidogenic organs, we visualize the innervation of steroidogenic organs as well as steroidogenic target cells such as germline stem cells.

Protocols for Visualizing Steroidogenic Organs and Their Interactive Organs with Immunostaining in the Fruit Fly Drosophila melanogaster

The Drosophila ovary is an excellent model system for studying stem cell niche development. Though methods for dissecting larval and adult ovaries have been published, pupal ovary dissections require different techniques that have not been published in detail. Here we outline a protocol for dissecting, staining, and mounting pupal ovaries.

Dissection and Staining of Drosophila Pupal Ovaries

The basement membrane is essential for tissue and organ morphogenesis during development. To better understand the mechanisms leading to proper placement of this structure, the protocol presented describes methods to visualize and characterize the intracellular trafficking and secretion of basement membrane proteins in epithelial cells using confocal and super-resolution microscopy.

Confocal and Super-Resolution Imaging of Polarized Intracellular Trafficking and Secretion of Basement Membrane Proteins During Drosophila Oogenesis

Here, we provide a detailed protocol to perform live imaging of the asymmetric division of germline stem cells (GSCs) in the Drosophila ovarian niche. We use a transgenic line that ubiquitously expresses a green fluorescent protein (GFP) fusion of the spectrosome protein Par-1.

Extended Live Imaging of Female Drosophila melanogaster Germline Stem Cell Niches

52.3K Views.  Princeton University. Dissection of Drosophila Ovaries