Lumplan n

Manufactured by Olympus

The LUMPlan N is a series of infinity-corrected objectives designed for biological and material science applications. The objectives feature high numerical aperture and long working distance to enable high-resolution imaging. They are compatible with standard laboratory microscopes.

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Lab products found in correlation

Fluo 4 am, by Thermo Fisher Scientific (3 mentions) Neo scmos camera, by Oxford Instruments (2 mentions) Collagenase type 2, by Thermo Fisher Scientific (2 mentions) Pluronic f 127, by Thermo Fisher Scientific (2 mentions) Probenecid, by Thermo Fisher Scientific (2 mentions) Dispase, by Thermo Fisher Scientific (2 mentions) Bx51wi fixed stage microscope, by Olympus (2 mentions) Daf fm, by Thermo Fisher Scientific (1 mentions) Neo scmos digital camera, by Oxford Instruments (1 mentions) Nis element, by Nikon (1 mentions) Zyla scmos camera, by Oxford Instruments (1 mentions) Metamorph, by Molecular Devices (1 mentions)

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LUMPlan (2 citations)

3 protocols using lumplan n

Intracellular Calcium and Nitric Oxide Imaging

Cited 1 time

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Live LMMP whole mount preparations were loaded with 4 μM Fluo-4-AM or DAF-FM [4-amino-5-methylamino-2′,7′-difluorofluorescein] (Life Technologies) for 45 minutes at 37°C as described elsewhere⁴ (link) to measure intracellular Ca²⁺ and NO, respectively. The loaded tissue was continuously perfused with prewarmed buffer (34°C, 3 mL/min) and drugs were bath applied. Images were acquired every 1–5 seconds with a Neo sCMOS digital camera (Andor, South Windsor, CT) through the 40× water-immersion objective (LUMPlan N, 0.8 n.a.) of an upright Olympus BX51W1 fixed-stage microscope controlled by Andor IQ3 software.

Brown I.A., McClain J.L., Watson R.E., Patel B.A, & Gulbransen B.D. (2015). Enteric Glia Mediate Neuron Death in Colitis Through Purinergic Pathways That Require Connexin-43 and Nitric Oxide. Cellular and Molecular Gastroenterology and Hepatology, 2(1), 77-91.

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Calcium Imaging of Intestinal Motility

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Live whole-mounts of the ileal and colonic MP were prepared for Ca²⁺ imaging as described by Fried and Gulbransen ²⁶. Briefly, distal ileal and colonic segments were collected in ice-cold DMEM and transferred to sylgard-coated, open diamond shaped bath recording chambers and then opened along the mesenteric border, pinned flat and microdissected. LMMP preparations were incubated for 15 min at room temperature in an enzyme mixture consisting of 150 U/ml Collagenase type II and 1 U/ml Dispase (Life Technologies) dissolved in DMEM before gentle trituration. LMMPs were loaded in the dark for 45 min at 37°C (5% CO_2, 95% air) with 4 μM Fluo-4 AM, 0.02% Pluronic F-127 and 200 μM water-soluble Probenecid (Life Technologies) in DMEM. LMMPs were washed three times with DMEM and incubated with 200 μM Probenecid in DMEM 15 min to de-esterify before imaging. Images were acquired every 1–2 seconds (s) through the 40X water-immersion objective (LUMPlan N, 0.8 n.a.) of an upright Olympus BX51WI fixed-stage microscope (Olympus) using IQ2 software and a Neo sCMOS camera (Andor, South Windsor, CT). Whole mounts were superfused with Krebs buffer (37° C) at 2–3 mL min ⁻¹.

McClain J.L., Fried D.E, & Gulbransen B.D. (2015). Agonist-Evoked Ca2+ Signaling in Enteric Glia Drives Neural Programs That Regulate Intestinal Motility in Mice. Cellular and Molecular Gastroenterology and Hepatology, 1(6), 631-645.

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Imaging Colonic Neuromuscular Preparations

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LMMP whole-mount preparations were microdissected from mouse colons and incubated for 15 minutes at room temperature in an enzyme mixture consisting of 150 U/mL collagenase type II and 1 U/mL dispase (Life Technologies, Carlsbad, CA). Samples from C57BL/6 and Sox10::CreER^T2+/-/Cx43^f/f mice were loaded in the dark with 4 μmol/L Fluo-4 AM, 0.02% Pluronic F-127, and 200 μmol/L probenecid (Life Technologies) in DMEM/F-12 at 37°C.¹⁴ Images were acquired every 1–2 seconds through the 40× water immersion objective (LUMPlan N; numeric aperture, 0.8) of an upright Olympus BX51WI fixed-stage microscope (Olympus) using IQ3 software (Andor Technology Ltd, Belfast, UK), MetaMorph (Molecular Devices, San Jose, CA), or NIS Elements (Nikon Instruments Inc, Melville, NY) software and a Neo sCMOS camera or a Zyla sCMOS camera (Andor Technology Ltd). We continually perfused whole mounts at 2–3 mL/min with buffer using a gravity flow perfusion system. Agonists were dissolved in buffer and bath applied for 30 seconds. Antagonists were dissolved in buffer and applied for either 20 minutes or 3 minutes before imaging.

Delvalle N.M., Dharshika C., Morales-Soto W., Fried D.E., Gaudette L, & Gulbransen B.D. (2018). Communication Between Enteric Neurons, Glia, and Nociceptors Underlies the Effects of Tachykinins on Neuroinflammation. Cellular and Molecular Gastroenterology and Hepatology, 6(3), 321-344.

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