C9903

Manufactured by Merck Group

Sourced in United States

The C9903 is a laboratory equipment product manufactured by Merck Group. It is designed for general laboratory use. The core function of the C9903 is to perform specific tasks required in a laboratory setting. Detailed description of the product's intended use or interpretation of its capabilities is not available.

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Lab products found in correlation

Isoflurane cp, by CP-Pharma (1 mentions) Metacam, by Boehringer Ingelheim (1 mentions) Fast green dye, by Merck Group (1 mentions) Pulled glass micropipette, by Drummond (1 mentions) Dmra2 compound microscope, by Leica (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Similac advance powder, by Abbott (1 mentions)

5 protocols using c9903

Visualising Avian Visual Cortex Projections

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Distal fibre terminals of V1 within the upper beak were visualized by neuronal tract tracing. Birds were anaesthetized with Isoflurane CP® (~ 1–1.5% Vol. dissolved in oxygen; 1 ml/ml; cp-pharma, Burgdorf, Germany). V1 was accessed unilaterally through an incision along the dorsal rim of the orbit and careful retraction of the eyeball and oculomotor muscles. This procedure was identical to previous studies^{6 (link),17 (link),18 (link),25 (link)–27 (link),29 (link)}. Approx. 250 nl of the neuronal tracer substance Cholera toxin subunit B (CtB; 1% in distilled water; C9903, Sigma-Aldrich, St. Louis, MO, USA) was administered by pressure injection into the nerve using a microinjector (WPI-2000, World Precision Instruments, Sarasota, FL, USA) and bevelled glass capillaries. After the injection, all tissues were repositioned and resealed using cyanoacrylate surgical glue (Histoacryl®, BRAUN, Rubi, Spain). For post-surgical analgesia, each bird was administered meloxicam (Metacam®, Boehringer Ingelheim, Ingelheim, Germany), 0.1 ml/kg body weight dissolved in 0.9% sodium chloride (NaCl), intramuscular 24- and 48-h post-surgery. Each bird was given three to six days to recover from the surgery and to let the tracer transport.

Curdt F., Haase K., Ziegenbalg L., Greb H., Heyers D, & Winklhofer M. (2022). Prussian blue technique is prone to yield false negative results in magnetoreception research. Scientific Reports, 12, 8803.

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Retrograde Labeling of Cutaneous and Muscle Afferents

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One week prior to perfusion, animals were sedated with ketamine (10mg/kg for cutaneous injections & 5mg/kg for muscle injections), and Cholera Toxin subunit B (CT-B, 10 μl, Sigma C9903, 1% concentration) was injected bilaterally using a Hamilton syringe. Digit pad injections were made subcutaneously into the distal and middle finger pads of digits 1-3 (M1603; Table 1, and Fig. 1B), to label cutaneous afferents. Each injection was targeted to the center of the pad, and the needle was held in the same plane as the skin and gentle rotated until it penetrated the skin so we could ensure it did not reach deep tissue. Total injected volume of CT-B in each hand was 60 μl (2 injections per digit). Muscle injections were made bilaterally into the first dorsal interosseous (FDI; 3 x 10 μl) and abductor pollicis brevis (AbPb; 5 x 10 μl) muscles (M1804 and M1805; Fig. 1B). We used a lower dose of ketamine than usual in these animals; this was sufficient for sedation while optimising muscle tone in the hand to help guide placement of the injections.

Fisher K.M., Garner J.P, & Darian-Smith C. (2022). Small sensory spinal lesions that affect hand function in monkeys, greatly alter primary afferent and motor neuron connections in the cord. The Journal of comparative neurology, 530(17), 3039-3055.

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Retrograde Tracing of Cranial Motoneurons

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For CT and EO muscle injections, adult (age 6–8 weeks) Nts^Cre mice were used that had been previously injected as described above with AAVDJ-hSyn-FLEX-mGFP-2A-Synaptophysin-mRuby into RAm 8 weeks before the muscle injections. Mice were anesthetized with isoflurane (3% for induction and 1–2% for maintenance) and then pre-treated with analgesic (carprofen 5 mg kg⁻¹ and buprenorphine SR 0.5–1.0 mg kg⁻¹, subcutaneous). For CT injections, a 1-cm incision was made in the ventral neck, and the CT muscles were exposed by dissection of the overlying strap muscles. A pulled glass micropipette (Drummond Scientific, 5-000-2005) was then used to inject 200–300 nl of 1% CTB solution (Sigma-Aldrich, C9903, diluted in PBS + 0.05% Fast Green dye) into the left and right CT muscles. The overlying skin was sutured, and the mouse was placed in a heated recovery cage. For EO injections, 400–1,000 nl of 1% CTB was similarly injected into the left and right EO muscles through an incision in the overlying skin that was sutured after injection. Mice recovered for 3 d (CT) or 7 d (EO) before perfusion and immunostaining.

Veerakumar A., Head J.P, & Krasnow M.A. (2023). A brainstem circuit for phonation and volume control in mice. Nature Neuroscience, 26(12), 2122-2130.

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Retrograde Tracing of Motor Neuron Pools

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Hypoglossal and phrenic motor neurons were labeled with the retrograde tracer, cholera toxin subunit β (CT-β), as previously described^{23 (link),24 (link)}. A 0.2% solution of CT-β (Millipore-Sigma, C9903) in Lactated Ringers was delivered through intralingual and intrapleural injections to 12 mo old mice (N = 2 – 3 per tissue per genotype). 72 hours after injection, the brainstem and spinal cords en bloc within the bone were harvested and fixed in 4% paraformaldehyde (PFA) for 24 hours. The brainstems and spinal cords were extracted from the soft tissue and bone, then fixed in 4% PFA for an additional 24 hours. Extracted brainstems and spinal cords were then cryopreserved in 30% sucrose then embedded in OCT. The medulla and cervical spinal cord were sectioned at 40 μm. Every second section from the hypoglossal region of the medulla and from C3–C5 within the cervical spinal cord was stained with an anti-cholera toxin antibody (List-Biologicals, 703). A biotinylated secondary antibody (VECTASTAIN ABC kit, Vector Laboratories, 1:200) exposed with 3,3’-diaminobenzidine (DAB) was applied. Sections were counterstained with cresyl violet, then imaged with a Leica DMRA2 Compound Microscope with Open Lab Software at 10x magnification. Motor neurons in the XII and phrenic motor pools were independently counted by two lab members who were blinded to the genotype of the mice.

McCall A.L., Dhindsa J.S., Pucci L.A., Kahn A.F., Fusco A.F., Biswas D.D., Strickland L.M., Tseng H.C, & ElMallah M.K. (2020). Respiratory pathology in the Optn−/− mouse model of Amyotrophic Lateral Sclerosis. Respiratory physiology & neurobiology, 282, 103525.

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Piglet Immunization and Dietary Intervention Study

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The animal study was conducted in accordance with the ethical guidelines for animal research approved by the Institutional Animal Care and Use Committee at the University of Arkansas for Medical Sciences. The detailed experimental design as well as the diet composition were previously published (19 (link)). Briefly, White Dutch Landrace Duroc male piglets within 2-d old were randomly assigned to two groups (n = 26/group), fed an isocaloric diet of HM (Mother’s Milk Bank of North Texas), or a dairy-based MF (milk formula; Similac Advance powder; Ross products, Abbott Laboratories, Columbus, OH) to meet the nutrient requirements of growing pigs as per the guidelines published by the National Research Council (NRC) (27 ). At postnatal day (PND) 14 complementary food (i.e., solid pellets) (starter pellets; Teklad, TD 140608; Harlan Laboratories) was introduced to the piglets and weaned to ad libitum solid pellets from PND21 to PND51 (19 (link)). Piglets were immunized on PND 21 and PND 35 with oral administration of 100 µg of cholera toxin (C8052, Millipore Sigma) and 100 µg of cholera toxin subunit B (CTB; C9903, Millipore Sigma). Piglets also received The DAPTACEL [diphtheria, tetanus, pertussis (DTaP)] vaccine (0.5 mL; Arkansas Children’s Hospital pharmacy) by intramuscular injection. Control piglets received vehicle.

Rosa F., Matazel K.S., Bowlin A.K., Williams K.D., Elolimy A.A., Adams S.H., Bode L, & Yeruva L. (2020). Neonatal Diet Impacts the Large Intestine Luminal Metabolome at Weaning and Post-Weaning in Piglets Fed Formula or Human Milk. Frontiers in Immunology, 11, 607609.

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