Microinjection of Drosophila Nurse Cells: A Method of Intracellular Delivery

Published: April 30, 2023

Abstract

Source: Catrina, I. E., et al. Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers. J. Vis. Exp. (2019).

Due to their large size and relatively simple organization, Drosophila nurse cells are ideally suited for live cell imaging studies. This video describes a microinjection method for exogenous compounds delivery into the cells, and the featured protocol demonstrates the procedure with fluorescent oligonucleotide probes called molecular beacons (MBs) used to visualize endogenous mRNA transcripts.

Protocol

This protocol is excerpted from Catrina et al., Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers, J. Vis. Exp. (2019). 1. Design of MBs for Live Cell Imaging Fold the target RNA sequence to predict the mRNA target's secondary structure using the "RNA form" from the mfold server (…

Materials

Spectrofluorometer Fluoromax-4 Horiba-Jobin Yvon n/a Photon counting spectrofluorometer
Quartz cuvette Fireflysci (former Precision Cells Inc.) 701MFL
Dumont #5 tweezer World Precision Instruments 501985 Thin tweezers are very important to separate out the individual egg chambers
Halocarbon oil 700 Sigma-Aldrich H8898
Cover slip No.1 22 mm x 40 mm VWR 48393-048
Dissecting microscope Leica MZ6 Leica Microsystems Inc. n/a
CO2 fruit fly anesthesia pad Genesee Scienific 59-114
Tris-HCL pH 7.5 Sigma-Aldrich 1185-53-1
Magnesium chloride Sigma-Aldrich 7791-18-6
NaCl Sigma-Aldrich 7647-14-5
Spinning disc confocal microscope Leica DMI-4000B inverted microscope equipped with Yokogawa CSU 10 spinning disc Leica Microsystems Inc. n/a
Hamamatsu C9100-13 ImagEM EMCCD camera Hamamatsu n/a
PatchMan NP 2 Micromanipulator Eppendorf Inc. 920000037
FemtoJet Microinjector Eppendorf Inc. 920010504
Injection needle: Femtotips II Eppendorf Inc. 930000043
Loading tip: 20 μL Microloader Eppendorf Inc. 930001007
Micro Cover glasses no. 1 or 1.5, 22 mm x 40 mm VWR 48393-026; 48393-172
Dry yeast Any grocery store n/a
Computer, > 20 GB RAM Although processing can be carried out on most computers, higher capabilities will increase the speed of the processing

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Cite This Article
Microinjection of Drosophila Nurse Cells: A Method of Intracellular Delivery. J. Vis. Exp. (Pending Publication), e20109, doi: (2023).

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