Microinjection Setup
Microinjection Setup - Related Videos
08:12
Zebrafish Microinjection Techniques
0 Views •
2023
One of the major advantages to working with zebrafish (Danio rerio) is that their genetics can be easily manipulated by microinjection of early stage embryos. Using this technique, solutions containing genetic material or knockdown constructs are delivered into the blastomeres: the embryonic cells sitting atop the yolk of the newly fertilized egg. Delivery into the cytoplasm is achieved either through direct injection into the blastomere, or via natural cytoplasmic movements that occur after a...
05:32
Embryo Microinjection for Transgenesis in Drosophila
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2024
This article takes the phiC31 integrase-mediated transgenesis in Drosophila as an example and presents an optimized protocol for embryo microinjection, a crucial step for creating transgenic flies.
13:39
Intranuclear Microinjection of DNA into Dissociated Adult Mammalian Neurons
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Cited by 20 •
2009
Direct intranuclear injection of cDNA is an effective transfection technique for post-mitotic cells. This method provides high levels of heterologous protein expression from single or multiple cDNA constructs and enables protein function to be studied in a physiologically relevant environment with a variety of single cell assays.
12:10
Microinjection of Xenopus Laevis Oocytes
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Cited by 14 •
2009
Here we demonstrate cytoplasmic microinjection of Xenopus laevis oocytes with a nuclear import substrate, as well as preparation of the injected oocytes for visualization by thin-sectioning electron microscopy.
10:06
Personalized Needles for Microinjections in the Rodent Brain
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Cited by 3 •
2018
We describe here a protocol for microinjection in the rodent brain that uses quartz needles. These needles do not produce detectable tissue damage and ensure reliable delivery even in deep regions. Moreover, they can be adapted to research needs by personalized designs and can be re-used.
FAQs
Reliable delivery depends on coordinating the microscope view, micromanipulators, pipettes, and pressure controls as one system. Micromanipulators provide the fine positional adjustments needed to bring the injection pipette to the target, while microscopic guidance supports accurate penetration. This coordination matters because small positioning errors can affect delivery precision and the survival of the treated cell or embryo.
The holding pipette and injection pipette perform complementary tasks. The holding pipette stabilizes the specimen so it remains positioned during manipulation; the narrower injection pipette then penetrates the cell membrane and delivers the selected material. Separating stabilization from delivery allows the operator to control specimen movement and injection placement independently, which is important when working with individual cells or embryos.
Pressure control determines how material moves through the injection pipette and helps regulate the delivered volume. Excessive force or rough handling can compromise the specimen, whereas controlled pressure supports a balance between effective delivery and survival. This balance is especially important when introducing DNA, RNA, proteins, or other reagents whose cellular effects are being examined.
A typical workflow begins by aligning the microscope, micromanipulators, pipettes, and pressure controls, then positioning the specimen with the holding pipette. The injection pipette is brought into place under microscopic guidance, introduced through the cell membrane, and used to deliver the chosen material. The operator then considers consistency and specimen survival, both central indicators of a successful setup.
Calibration and alignment should be established before repeated injections rather than treated as optional adjustments. Calibration helps the operator work with controlled pressure and consistent pipette movement, while alignment keeps the injection path accurately related to the microscope view and specimen. Together, these preparations reduce variation between injections and make cellular responses easier to interpret.
This approach is useful when an experimental question requires material delivery to one cell or embryo rather than to a whole population. Applications include gene-function studies, cell-development research, fertilization studies, and transgenesis. Because delivery is localized, researchers can examine responses that bulk delivery methods may obscure at the level of individual specimens.