Nanoject 2 injector

Manufactured by Drummond

Sourced in United States

The Nanoject II injector is a laboratory equipment designed for the precise injection of small volumes of liquids. It features an automated injection system capable of delivering nanoliter-scale volumes with high accuracy and repeatability. The core function of the Nanoject II is to provide a reliable and precise method for the controlled delivery of samples, reagents, or other liquids in various laboratory applications.

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Lab products found in correlation

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Close spelling variants of this product

Nanoject II (397 citations) Nanoject II Auto-Nanoliter Injector (80 citations) Nanoject injector (28 citations) Nanoject II™ Nanoliter injector (5 citations) Nanoject II automatic nanoliter injector (3 citations) Nanoject II automatic injector (2 citations)

55 protocols using nanoject 2 injector

Mosquito Ookinete Injection Protocol

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After ookinete purification from in vitro culture described above, each of 40 mosquitoes (4 to 5 days old) was injected intrathoracically with 138 nl of PBS containing 690 ookinetes using a Nanoject II injector (Drummond Scientific, USA). The hemocoel oocysts within the mosquitoes were observed under the stereoscopic fluorescence microscope (M165FC, Leica) at indicated times after injection.

Yang Z., Shi Y., Cui H., Yang S., Gao H, & Yuan J. (2021). A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission. Science Advances, 7(30), eabf6015.

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Dengue Virus Infection in Mosquitoes

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Female mosquitoes at 3–5 days post-emergence were anesthetized by CO₂. Each mosquito was intrathoracically inoculated with 69 nl of DENV-2 16681 (6.9 × 10² PFU in 69 nl) grown in C6/36 cell culture using a Nanoject II injector (Drummond, Broomall, Pennsylvania, USA) as described previously [19 (link)]. Mosquitoes were then transferred into cylindrical containers fitted with nylon mesh on the top. Saliva was collected from infected mosquitoes on day 7 after infection. Viral titer was determined by plaque assay [19 (link)].

Cheang K.W., Chen W.Y., Wu-Hsieh B.A, & Shiao S.H. (2021). Infecting mosquitoes alters DENV-2 characteristics and enhances hemorrhage-induction potential in Stat1-/- mice. PLoS Neglected Tropical Diseases, 15(8), e0009728.

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Fly-based IIV-6 Infection Protocol

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Fly stocks were grown on standard cornmeal-agar molasses at 25°C. w[1118] strain was used as wildtype flies. Loquacious transgenic flies were introduced elsewhere (Fukunaga et al., 2012 (link)). Two transgenic lines expressing loqs specific isoforms were used here: Loqs-PD deficient (w[1118];loqs[KO]/CyO;P{w+,FLAG-Loqs-PB}/TM3,Sb[1]) and the Control (w[1118];loqs[KO]/CyO;P{w+,FLAG-Loqs-PB-PD}/TM3,Sb[1]) (Fukunaga et al., 2012 (link)). For IIV-6 infections, 4 to 6-day-old female flies were intrathoracic injected (Nanoject II injector; Drummond Scientific) with 5 X 10⁵ TCID₅₀ (w[1118] flies for SOLiD platform sequencing) or 5,000 TCID₅₀ (Control and Loqs-PD deficient flies for Illumina platform sequencing) diluted in 69 nL of non-supplemented Schneider’s medium. The flies were incubated at 25°C and harvested in TRIzol reagent (Invitrogen). Samples were homogenized in the Mini-Beadbeater homogenizer (BioSpec). RNA and DNA were extracted according to TRIzol’s protocol.

Tang H., Severinov K., Goldfarb A, & Ebright R.H. (1995). Rapid RNA polymerase genetics: one-day, no-column preparation of reconstituted recombinant Escherichia coli RNA polymerase.. Proceedings of the National Academy of Sciences of the United States of America, 92(11), 4902-4906.

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Stereotaxic AAV-mediated transgene delivery

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Adeno-associated virus (AAV) (~5×10¹² gc/ml) encoding murine preproSST transgene fused with GFP via T2A self-cleavage spacer sequence was prepared by AAVpro purification kit (TaKaRa Bio, Japan) and bilaterally injected into PFC (AP +1.9 mm; ML ±0.15 mm; DV −0.8 mm), using a stereotaxic apparatus (RWD Life Science, USA). A total of 0.5 µl of purified virus was delivered on each hemisphere over a 5-min period using glass microcapillary pipettes (1.14 mm O.D. x 3.5” length x 0.53mm I.D.) and the Nanoject II injector (Drummond, PA, USA). Mice (8 to 10 weeks of age) were anesthetized by isoflurane inhalation during the stereotaxic injection, and used for downstream assays 3 weeks post-injection.

Tomoda T., Sumitomo A., Newton D, & Sibille E. (2022). Molecular origin of somatostatin-positive neuron vulnerability. Molecular psychiatry, 27(4), 2304-2314.

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Gene Silencing in Anopheles stephensi

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All the dsRNA products for silencing genes such as TEP1, Rel1, GNBP-B1, and NOX5 were generated using a cDNA template from A. stephensi, and the MEGAscript^TM RNAi Kit (Thermo Fisher Scientific) as previously described^{3 (link),7 (link)}. In brief, 3-day-old female A. stephensi were cold-anesthetized and injected with 3 mg/mL of dsRNA using a Nanoject II injector (Drummond Scientific Co., Broomall, PA, USA), 2 days prior to infection. Silencing efficiency was examined at day 2 post-knockdown by qPCR, as described above. Primers designed for specific transcript targets are listed in Supplementary Table 3.

Zhu F., Zheng H., Chen S., Zhang K., Qin X., Zhang J., liu T., Fan Y., Wang L., Li X., Zhang J, & Xu W. (2022). Malaria oocysts require circumsporozoite protein to evade mosquito immunity. Nature Communications, 13, 3208.

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Clodronate Liposomes Depletion of Mosquito Hemocytes

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The day prior to CSP_mut infection, naïve female mosquitoes (3–5 days old) were injected intrathoracically with either 69 nL of control liposomes or CLDs (Standard Macrophage Depletion Kit; Clodrosome^® + Encapsome^®; Encapsula Nano Sciences, Brentwood, TN, USA) using a Nanoject II injector (Drummond Scientific Co.). As previously described^{22 (link)}, 1:5 dilutions of liposomes and CLDs in 1 × PBS were used for the experiments. Following liposome injection, mosquitoes were infected with CSP_mut parasites. The effect of clodronate liposomes on the depletion of mosquito hemocytes was determined by the expression of cell markers NimB2 (hemocytes), LRIM15 (granulocytes), and SCRB9 (oenocytoids) with real-time PCR as previously reported^{26 (link)}. The change in oocyst melanization, the number of sporozoites, and relative expression of TEP1 were determined as described above.

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Microinjection of Plasmid DNA into Xenopus

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Glass capillaries (3.5-inch length, 0.02-inch width: Drummond Scientific) were pulled (Sutter Instrument Co P-97) and the tip was broken under a dissecting scope using forceps. We injected a solution of 0.25–0.27 μg/μl of purified plasmid DNA (pCMV-GFP, Addgene #11153) mixed with 0.01% Fast Green using a Nanoject II injector (Drummond Scientific). Injection volumes are listed below with the corresponding protocol. We used plasmid concentrations based on that reported for X. laevis [3 (link), 16 , 46 ].

Delia J., Gaines-Richardson M., Ludington S.C., Akbari N., Vasek C., Shaykevich D, & O’Connell L.A. (2023). Tissue-specific in vivo transformation of plasmid DNA in Neotropical tadpoles using electroporation. PLOS ONE, 18(8), e0289361.

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Targeted dsRNA Depletion of Mosquito Proteins

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To deplete TEP1, LRIM1, TEP1*S, or TEP1*R, dsRNAs for TEP1, LRIM1, LacZ, TEP1*S (dsS), and TEP1*R (dsR) were produced from plasmids containing the dsRNA-target sequence flanked by two T7 promoters [5 (link),21 (link),44 ]. To deplete HPX2, dsRNA for HPX2 was produced from PCR-amplified fragments flanked with two T7 promoters as described [7 (link)]. RNA synthesis and purification were performed using MegaScript and MegaClear kits (Ambion). RNA concentrations were measured by Nanodrop (Thermo Scientific) before annealing 3 μg/μl of sense and anti-sense RNAs by boiling. DsRNAs were injected (69 nl) into the thorax of CO₂-immobilized 1-d-old mosquitoes using a glass capillary mounted onto a Nanoject II injector (Drummond).

Pompon J, & Levashina E.A. (2015). A New Role of the Mosquito Complement-like Cascade in Male Fertility in Anopheles gambiae. PLoS Biology, 13(9), e1002255.

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Injection of Flies with ThT and PGN

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In Figure 5C, and in related Figure S2, male and female flies were injected with 64 nL of the following solutions using Nanoject II injector (Drummond Scientific): untreated: no injection; DMSO: 5% DMSO in sterile Dulbecco’s modified PBS (Corning); ThT: 1 mM ThT, 5% DMSO in PBS; DMSO+PGN: 5% DMSO, 0.5 mg/ml E. coli strain 1106 PGN in PBS; ThT+PGN: 1 mM ThT, 0.5 mg/ml PGN in PBS. In Figure 5D, male flies were injected with 64 nL of indicated concentrations of ThT diluted in 5% DMSO in PBS, +/− 0.5 mg/ml PGN. Flies were harvested 1 h post-injection, snap-frozen on dry-ice, homogenized in TRIzol® reagent (Invitrogen), and total RNAs were extracted according to the manufacturer’s protocol. In Figure 5E, and in related Figure S3, flies were injected and harvested as described above, but 40 μM TCT or 400 μM TCT was used instead of PGN.

Kleino A., Ramia N.F., Bozkurt G., Shen Y., Nailwal H., Huang J., Napetschnig J., Gangloff M., Chan F.K., Wu H., Li J, & Silverman N. (2017). Peptidoglycan-Sensing Receptors Trigger the Formation of Functional Amyloids of the Adaptor Protein Imd to Initiate Drosophila NF-κB Signaling. Immunity, 47(4), 635-647.e6.

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AAV-Mediated Somatostatin Overexpression in PFC

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Adeno-associated virus (AAV) (~5x10 12 gc/ml) encoding murine preproSST transgene fused with GFP via T2A self-cleavage spacer sequence was prepared by AAVpro purification kit (TaKaRa Bio, Japan) and bilaterally injected into PFC (AP +1.9 mm; ML ±0.15 mm; DV -0.8 mm), using a stereotaxic apparatus (RWD Life Science, USA). A total of 0.5 µl of purified virus was delivered on each hemisphere over a 5-min period using glass microcapillary pipettes (1.14 mm O.D. x 3.5" length x 0.53mm I.D.) and the Nanoject II injector (Drummond, PA, USA). Mice (8 to 10 weeks of age) were anesthetized by isoflurane inhalation during the stereotaxic injection, and used for downstream assays 3 weeks post-injection.

Tomoda T., Sumitomo A., Newton D.F., & Sibille E. (2021). Molecular origin of somatostatin-positive neuron vulnerability.

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