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Femtojet 4

Manufactured by Eppendorf
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Sourced in Germany

The FemtoJet 4 is a microinjector designed for precise and controlled injection of small liquid volumes into biological samples. It features an advanced pneumatic system to deliver femtoliter to microliter range injections with high accuracy and reproducibility.

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Lab products found in correlation

P 1000, by Sutter Instruments (2 mentions) Nitex mesh, by Genesee Scientific (2 mentions) Eg 40, by Narishige (2 mentions) Injectman 4, by Eppendorf (2 mentions) Pc 10, by Narishige (1 mentions) Mn 153, by Narishige (1 mentions) Evos m7000 imaging system, by Thermo Fisher Scientific (1 mentions) Celltracker cm dii dye, by Thermo Fisher Scientific (1 mentions) Pc 100, by Narishige (1 mentions) Purelink expi endotoxin free maxi plasmid purification kit, by Thermo Fisher Scientific (1 mentions) Sigmacote, by Merck Group (1 mentions) Protamine coated glass bottom dishes, by MatTek (1 mentions)

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FemtoJet (220 citations)

8 protocols using femtojet 4

1

Microinjection of Beads into Unfertilized Eggs

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Unfertilized eggs were placed on protamine-coated 50-mm glass-bottom dishes (MatTek Corporation) after removing the jelly coat through a 80-μm Nitex mesh (Genesee Scientific). The bead suspensions were injected using a micro-injection system (FemtoJet 4; Eppendorf) and a micro-manipulator (Injectman 4; Eppendorf). Injection pipettes were prepared from siliconized (Sigmacote) borosilicate glass capillaries (1 mm diameter). Glass capillaries were pulled using a needle puller (P-1000; Sutter Instrument) and ground with a 40° angle on a diamond grinder (EG-40; Narishige) to obtain a 10-μm aperture. Injection pipettes were back-loaded with ~2 μL bead suspension before each experiment, and were not reused.
Najafi J., Dmitrieff S, & Minc N. (2023). Size- and position-dependent cytoplasm viscoelasticity through hydrodynamic interactions with the cell surface. Proceedings of the National Academy of Sciences of the United States of America, 120(9), e2216839120.
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2

Microinjection of Etoposide into Embryos

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Glass microcapillaries were drawn on a needle puller (PC-10; Narishige, Tokyo, Japan), and solution of 20 µM etoposide, 0.2 M KCl, and 1% Rhodamine dextran (MW 10,000) was Microinjected into embryos. Rhodamine was used for visual control of the delivery of solution during injection via the appearance of red color under the stereomicroscope and again during sorting under a fluorescence stereomicroscope. Microcapillaries loaded with solution were mounted on a micromanipulator (MN-153, Narishige, Japan) attached to a pneumatic injector (FemtoJet® 4×, Eppendorf, Hamburg, Germany). Non-dechorionated embryos were placed in a Petri dish coated with 1% agar. Tips of the microcapillaries were carefully broken with forceps. The duration and pressure of injection pulses were determined using the oil droplet method and kept constant over the experiment while ~10 nL was injected into the animal pole of each embryo.
Gazo I., Franěk R., Šindelka R., Lebeda I., Shivaramu S., Pšenička M, & Steinbach C. (2020). Ancient Sturgeons Possess Effective DNA Repair Mechanisms: Influence of Model Genotoxicants on Embryo Development of Sterlet, Acipenser ruthenus. International Journal of Molecular Sciences, 22(1), 6.
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3

Tick Bacterial Infection Assay

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Log-phase bacteria were diluted in PBS to a concentration of 107 CFU/mL and injected into I. scapularis nymphs using an Eppendorf FemtoJet 4x microinjector. At varying times post injection, ticks were crushed in PBS and plated on appropriate solid media to determine CFU. Injections of PBS alone was used as a monitor of tick fitness and survival of the injection trauma. To assess tick viability, ticks were monitored every hour after injection for movement and reaction to CO2 (human breath). Ticks that had curled their front legs, did not respond to CO2, and were not mobile even after picking up with forceps (no reflex straightening of legs) were considered dead.
Hayes B.M., Radkov A.D., Yarza F., Flores S., Kim J., Zhao Z., Lexa K.W., Marnin L., Biboy J., Bowcut V., Vollmer W., Pedra J.H, & Chou S. (2020). Ticks Resist Skin Commensals with Immune Factor of Bacterial Origin. Cell, 183(6), 1562-1571.e12.
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4

In Vivo Tumor Cell Tracking in Zebrafish

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At 48 h post-fertilization (hpf), dechorionated larvae were anesthetized with tricaine methanesulphonate (MS-222) and positioned on a modified Petri dish coated with 1.5% w/v agarose. Before injection, cells were labeled in vitro with CellTracker™ CM-DiI Dye (Thermo Fisher Scientific). Approximately 250 cells were resuspended in a complete medium, and 33.51 nL of cell solution were injected using commercially available ready-to-use tip needles (15 μm inner diameter, Eppendorf, Germany) into the yolk sac. An Eppendorf’s semi-automated microinjection system equipped with the micromanipulator InjectMan 4 connected to FemtoJet 4 × was used for the injections, with an injection pressure of 100 hPa, 0.2 s injection time and 20 hPa of compensation pressure. After injections, the larvae were immediately transferred into housing-keeping water. Injected larvae were kept at 28 ± 0.5 °C and examined every other day for monitoring tumor growth and invasion using the EVOS™ M7000 Imaging System (Thermo Fisher Scientific).
Talia M., Cirillo F., Spinelli A., Zicarelli A., Scordamaglia D., Muglia L., De Rosis S., Rigiracciolo D.C., Filippelli G., Perrotta I.D., Davoli M., De Rosa R., Macirella R., Brunelli E., Miglietta A.M., Nardo B., Tosoni D., Pece S., De Francesco E.M., Belfiore A., Maggiolini M, & Lappano R. (2023). The Ephrin tyrosine kinase a3 (EphA3) is a novel mediator of RAGE-prompted motility of breast cancer cells. Journal of Experimental & Clinical Cancer Research : CR, 42, 164.
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5

In Utero Myoblast Transplantation in Mice

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Injection pipettes were prepared according to a previous report [37 (link)]. Briefly, the glass tube (NARISHIGE, Tokyo, Japan) was formed thin using a pipette puller (PC-100, NARISHIGE), and the tip was formed into a needle shape using a diamond sharping wheel. 5.0 × 104 myoblasts or 2.0 × 104 ASCs were resuspended in 2 μl saline solution and loaded in injection pipette. Then, an injection pipette was connected to electronic microinjector (FemtoJet 4, Eppendorf, Hmburg, Germany). Pregnant mdx mice (E11.5) were anesthetized on a heater with inhalation of isoflurane and laparotomized. Placentas seen from outside the uterus were punctured with a pipette to a depth of 2 mm vertically and cells were injected with a microinjector (Fig. 1b). During the procedure, the uterus was moistened with warm saline to prevent dryness and loss of body temperature. After all fetuses were injected, the uterus was returned to the abdominal cavity and the abdomen was closed with a 4-0 braided absorbable suture. The transplanted mice were delivered by cesarean section at 19.5 days gestation and raised until 4 weeks of age for analysis.
Kihara Y., Tanaka Y., Ikeda M., Homma J., Takagi R., Ishigaki K., Yamanouchi K., Honda H., Nagata S, & Yamato M. (2022). In utero transplantation of myoblasts and adipose-derived mesenchymal stem cells to murine models of Duchenne muscular dystrophy does not lead to engraftment and frequently results in fetal death. Regenerative Therapy, 21, 486-493.
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6

Efficient Plasmid Microinjection for Culex Mosquitoes

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The injected DNA plasmids were prepared using PureLink Expi Endotoxin-Free Maxi Plasmid Purification Kit (Invitrogen, Cat.# A31231), aliquoted based on the concentrations outlined in the manuscripts, and later stored at −80 °C before proceeding to microinjection. All injections were performed on a microinjection station equipped with a FemtoJet 4 microinjector (Eppendorf). The prepared Cas9/sgRNA mixtures were injected into the posterior end of Culex quinquefasciatus embryos eggs freshly collected after oviposition (~1 h) to ensure efficient targeting of the germline.
Feng X., López Del Amo V., Mameli E., Lee M., Bishop A.L., Perrimon N, & Gantz V.M. (2021). Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes. Nature Communications, 12, 2960.
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7

Borosilicate Glass Capillary Microinjection

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Needles for microinjection were made by pulling Borosilicate Glass Capillaries (1B110F-4) (World precision instruments) using Narishige’s PC-100 micropipette puller with heater 1 set at 58 and no setting for heater 2. Injections were made using a FemtoJet® 4× (Eppendorf) with an injection pressure of 200 hPa.
Jensen H.H., Frantzen M.T., Wesseltoft J.L., Busuioc A.O., Møller K.V., Brohus M., Duun P.R., Nyegaard M., Overgaard M.T, & Olsen A. (2023). Human calmodulin mutations cause arrhythmia and affect neuronal function in C. elegans. Human Molecular Genetics, 32(12), 2068-2083.
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8

Microinjection of Unfertilized Eggs

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The jelly coat of unfertilized eggs was removed by passing them three times through an 80 μm Nitex mesh (Genesee Scientific) to facilitate egg adhesion on protamine-coated glass-bottom dishes (MatTek Corporation). Unfertilized eggs were transferred to protamine-coated glass-bottom dishes for a maximum time of 15 min before microinjection and fertilization. Microinjections were performed using a FemtoJet 4 and Injectman 4 micromanipulator (Eppendorf). Injection pipettes were prepared from siliconized (Sigmacote; Sigma-Aldrich) borosilicate glass capillaries (1 mm diameter). Glass capillaries were pulled with a needle puller (P-1000; Sutter Instruments) and ground with a 30° angle on a diamond grinder (EG-40; Narishige) to obtain a 5–10 μm aperture. Injection pipettes were back-loaded with 2 μl before each experiment and were not reused. Injection volumes were generally less than 5% of the egg volume, ~2–5 pL. GST-GFP-NLS and GST-mCherry-NLS proteins were diluted to ~2.5 mg/ml in PBS before injecting. Injecting undiluted GST-GFP-NLS did not affect developmental progression, suggesting that these protein concentrations are not detrimental to the embryo. RNA was diluted to ~50 ng/μl in PBS before injecting. These concentrations were empirically selected to optimize imaging while not affecting developmental progression.
Mukherjee R.N., Sallé J., Dmitrieff S., Nelson K.M., Oakey J., Minc N, & Levy D.L. (2020). The perinuclear ER scales nuclear size independently of cell size in early embryos. Developmental cell, 54(3), 395-409.e7.
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