Chorion and Vitelline Membrane Mechanical Removal: A Method to Prepare Drosophila Oocytes for Direct Observation

Abstract

Source: Perkins, A. T., Bickel, S. E. Using Fluorescence In Situ Hybridization (FISH) to Monitor the State of Arm Cohesion in Prometaphase and Metaphase I Drosophila Oocytes. J. Vis. Exp. (2017).

This video describes a method to mechanically remove the chorion and vitelline membranes from previously fixed mature Drosophila oocytes. The featured protocol shows a detailed demonstration of the procedure yielding oocytes compatible with fluorescent in situ hybridization assays.

Protocol

This protocol is an excerpt from Perkins and Bickel, Using Fluorescence In Situ Hybridization (FISH) to Monitor the State of Arm Cohesion in Prometaphase and Metaphase I Drosophila Oocytes, J. Vis. Exp. (2017). 1. Removal of Chorions and Vitelline Membranes NOTE: See Figure 1 for tools needed. Separate late stage oocytes<…

Representative Results

Figure 1: Cytology tools. Tools used in protocol: (1) pair of forceps (#5 Dumont); (2) tungsten needle; (3) deep well dish with cover; (4) shallow glass dissecting dish; (5) pulled Pasteur pipette. Please click here to view a larger version of this figure. <img alt="Figure 2" class="xfigimg" src="/…

Materials

Bovine serum albumin (BSA)	Fisher Scientific	BP1600-100	Prepare 10% stock Freeze aliquots
10% Triton X-100	Thermo Scientific	28314	Surfact-Amps
PBSBTx			1X PBS, 0.5% BSA, 0.1% Trition X-100
Forceps	Dumont		#5 INOX, Biologie
9" Disposable glass Pasteur pipettes	Fisher	13-678-20C	Autoclave to sterilize
Shallow glass dissecting dish			Custom made
Deep well dish (3 wells)	Pyrex	7223-34
Tungsten needle			homemade
Frosted glass slides, 25 x 75 mm	VWR Scientific	48312-002
15 mL conical tubes	Various
500 µl microfuge tubes	Various
Kimwipes	Various		disposable wipes