Calcein solution

Manufactured by Merck Group

Sourced in United States

Calcein solution is a fluorescent dye used in various laboratory applications. It is a water-soluble compound that emits a green fluorescence when excited by light of a specific wavelength. The core function of calcein solution is to serve as a versatile labeling agent for the detection and visualization of various biological and chemical substances in analytical and research settings.

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Lab products found in correlation

Avizo, by Thermo Fisher Scientific (1 mentions) Lsm 780 upright confocal microscope, by Zeiss (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Smz18 microscope, by Nikon (1 mentions) Chondrogenic medium, by Lonza (1 mentions) 5 bromo 4 chloro 3 indolyl phosphate bcip, by Merck Group (1 mentions) Isobutylmethylxanthine ibmx, by Merck Group (1 mentions) Dmem low glucose, by Merck Group (1 mentions) Oil red o solution, by Merck Group (1 mentions) Insulin, by Merck Group (1 mentions) Dexamethasone (dex), by Merck Group (1 mentions) Indomethacin, by Merck Group (1 mentions)

5 protocols using calcein solution

Calcein Staining of Zebrafish Larvae

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The larvae were immersed in a 0.2% calcein solution (Sigma-Aldrich) (pH 6.8) for 25 min and then washed three times with blue water. For in vivo observations, animals were anesthetized with 0.12% tricaine-metanesulfonate (MS222) in blue water. After mounting in methyl cellulose 5% (1.5%) plate, the larvae were observed using an epifluorescence stereomicroscope (Leica M167FC). Pictures were taken using Leica Application Suite imaging software version 3.7 (Leica, Wetzlar, Germany).
Confocal imaging was performed on a Zeiss LSM 780 upright confocal microscope (Carl Zeiss, Jena, Germany) with a W-Plan Apochromat × 20 objective, NA 1.0. The calcein staining was excited at 488 nm and the emission was collected at 492/577 nm. Z-stacks were acquired at 1.5 μm increments, every 1 min. Pictures were processed off-line using ImageJ (NIH) and Avizo (FEI).
4 zebrafish larvae were observed for each condition and time.

Silvent J., Akiva A., Brumfeld V., Reznikov N., Rechav K., Yaniv K., Addadi L, & Weiner S. (2017). Zebrafish skeleton development: High resolution micro-CT and FIB-SEM block surface serial imaging for phenotype identification. PLoS ONE, 12(12), e0177731.

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Shell Growth Rate in Giant Clams

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To study the impact of temperature and pCO₂ on shell growth rate, the mineralization front of giant clams was marked using calcein fluorochrome, which is irreversibly precipitated at the CaCO₃ mineralization site. Before the experiment starts, giant clams were immersed in a 100-mg.l⁻¹ calcein solution (Sigma Aldrich) (calcein diluted in 1-μm filtered-seawater) for 8 h in the dark. During the labelling procedure, the bath of calcein solution was aerated using bubblers and water current was created via pumps. Calcein-labelled specimens were then placed into the experimental tanks. At the end of the experiment, for each individual, a 5-mm thick section was cut along the maximal shell growth axis through the right valve using a Swap Top Inland® diamond saw. All sections obtained were polished and observed under epifluorescence with a Leitz Dialux® 22 microscope. The distance between the fluorescent calcein mark and the edge of the shell formed during the experiment was measured following the maximal growth direction. Daily shell extension rate (expressed in μm.day⁻¹) was obtained by dividing the measured distance by the number of days of incubation in experimental conditions.

Brahmi C., Chapron L., Le Moullac G., Soyez C., Beliaeff B., Lazareth C.E., Gaertner-Mazouni N, & Vidal-Dupiol J. (2021). Effects of elevated temperature and pCO2 on the respiration, biomineralization and photophysiology of the giant clam Tridacna maxima. Conservation Physiology, 9(1), coab041.

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Calcein Labeling for Bone Deposition

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Rats were given subcutaneous injection of calcein solution (8 mg/kg) (Sigma-Aldrich) at the 3^rd and 10^th day before their sacrifice. The jawbones were isolated, fixed and embedded. The embedded specimens of jawbones were cut into sections of 50 μm thickness, and then the calcein labeling was observed using a fluorescence microscope (Olympus). The distance between two adjacent green fluorescent bands was measured with Image Pro software to calculate new bone deposition rate of 7 days.

Shuai Y., Liu B., Rong L., Shao B., Chen B, & Jin L. (2022). OSGIN2 regulates osteogenesis of jawbone BMSCs in osteoporotic rats. BMC Molecular and Cell Biology, 23, 22.

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Analyzing Craniofacial Development in Zebrafish

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Live embryos previously growing in 0.003% PTU embryo media at ages 6 and 10 dpf were incubated in a 0.2% (w/v) calcein solution, pH 7.5 (Sigma-Aldrich, St. Louis, MO, USA), for 12 min followed by three 5-min washes in 0.003% PTU embryo media. Embryos were subsequently anesthetized in tricaine (see Zebrafish Husbandry) and mounted in 6% methylcellulose for imaging. Images were collected in the dark using a Nikon SMZ18 microscope equipped with a long-pass green filter (excitation 480 ± 40 nm; emission 510 nm). Significance testing for delayed craniofacial cartilage and bone at 6 dpf was calculated using Fisher’s exact test. Delay of cartilage development and bone calcification was characterized by decreased staining intensity, shape, and size of area (in the case of primary ossification centres), and the presence or lack thereof in comparison to their WT siblings in their respective experimental groups.

Hawkey-Noble A., Pater J.A., Kollipara R., Fitzgerald M., Maekawa A.S., Kovacs C.S., Young T.L, & French C.R. (2022). Mutation of foxl1 Results in Reduced Cartilage Markers in a Zebrafish Model of Otosclerosis. Genes, 13(7), 1107.

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Mesenchymal Differentiation Capacity Evaluation

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To confirm the mesenchymal activity of those cells that grew out naturally from the lipoaspirate clusters, while under culture conditions, their in vitro differentiation capacity was studied according to Noël et al. (26) . Briefly, 10 × 10 3 cells/ cm 2 were cultured in DMEM-low glucose supplemented with 10% FBS, 0.5 mM isobutyl-methyl xanthine (IBMX; Sigma-Aldrich), 200 μM indomethacin, 1 μM dexamethasone, and 10 μg/ml insulin (all from Sigma-Aldrich); after 2 weeks they were stained with fresh Oil red O solution (Sigma-Aldrich) for adipogenesis. After 3 weeks of culture in DMEM-low glucose supplemented with 10% FBS, 10 mM b-glycerophosphate, 0.2 mM ascorbic acid, and 10 nM dexamethasone (all from Sigma-Aldrich), for osteogenic differentiation, cells were stained with calcein solution (Sigma-Aldrich) to evidence mineralization (16) and with 5-bromo-4-chloro-3-indolyl phosphate (BCIP; Sigma-Aldrich) to evidence alkaline phosphatase activity (8). To demonstrate chondrogenic differentiation, pellets of 5 × 10 5 cells were cultured for 3 weeks in chondrogenic medium (Lonza, Cologne, Germany), formalin-fixed, embedded in paraffin, and immunostained for type II collagen by incubation with rabbit polyclonal anti-collagen type II primary antibody 1 mg/ml (Biogenesis, Oxford, UK) and with secondary antibody conjugated to peroxidase (Vector Laboratories).

Bosetti M., Borrone A., Follenzi A., Messaggio F., Tremolada C, & Cannas M. (2016). Human Lipoaspirate as Autologous Injectable Active Scaffold for One-Step Repair of Cartilage Defects. Cell transplantation, 25(6).

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