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Quantitative Microinjection Using Fluorescence Calibration of Streaming Microdroplets on a Superhydrophobic Surface

Quantitative Microinjection Using Fluorescence Calibration of Streaming Microdroplets on a Superhydrophobic Surface

Christopher L Moore, Erin M Taylor, Kelly K Ball, Laura J Bernock, Robert J Griffin, Seunghyun Jung, Amal Shoeib, Michael J Borrelli

Exp Cell Res. 2018 Sep 15;370(2):426-433.

PMID: 29981341

Abstract:

A simple and reproducible procedure was developed to measure the volume of liquid microinjected into cells. A calibration curve of droplet fluorescence intensity versus volume was constructed by injecting a fluorescent dextran solution through a 125-150 µm diameter micropipette into an oil-filled culture dish to create a spray of varied-sized droplets. The droplets retained a spherical shape because they were in an oil medium and they settled onto a glass surface coated with a superhydrophobic surface. Fluorescent micrographs of the droplets were obtained and analyzed with Image-J software to quantify the fluorescence intensity and radius of each spherical droplet to produce the calibration curve. Subsequently, Dut-145 human prostate carcinoma cells were microinjected with the same fluorescent dextran solution and fluorescent micrographs of the cells were obtained using the identical exposure conditions used to photograph the droplets. The measured fluorescence intensity of the microinjected cells was entered into the formula for the regression line that was fit to the calibration curve allowing determination of the volume of solution injected into each cell. Thus, a mixture consisting of known concentrations of a test material of test material (macromolecules, drugs, etc.) and a fluorescent dextran, volumetric, tracer can be used to quantify the relationship between the amount of a microinjected material and subsequent effects on cells.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP9004540-M	Dextran 70 Calibration		9004-54-0	Price

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