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Tricaine

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Tricaine is an aqueous solution containing the chemical compound tricaine methanesulfonate, which is commonly used as an anesthetic agent for fish and other aquatic organisms. It functions by reversibly depressing the central nervous system, allowing for safe and effective immobilization of the organism during procedures.

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Lab products found in correlation

Axio observer a1, by Zeiss (6 mentions) Eclipse 90i c1, by Nikon (2 mentions) Lsm700, by Nikon (2 mentions) 35 mm imaging dishes, by MatTek (2 mentions) Axiocam hrc camera, by Zeiss (2 mentions) Ic80 hd camera, by Leica camera (2 mentions) Nosepiece attachment, by Leica camera (2 mentions) M80 scope, by Leica camera (2 mentions) Low melting point agarose, by Thermo Fisher Scientific (1 mentions) Low melt agarose, by Thermo Fisher Scientific (1 mentions) Axiocam high, by Zeiss (1 mentions) Axiovision software, by Zeiss (1 mentions) Methylcellulose, by Merck Group (1 mentions) Alexa fluor 488 conjugated, by Thermo Fisher Scientific (1 mentions) Smz18, by Nikon (1 mentions) Pli 90, by Harvard Apparatus (1 mentions) Methyl cellulose solution, by Thermo Fisher Scientific (1 mentions) Tricaine, by Merck Group (1 mentions) Axiovision se64 rel 4, by Zeiss (1 mentions) Levamisole, by Thermo Fisher Scientific (1 mentions)

9 protocols using tricaine

1

Fluorescent Imaging of Zebrafish Embryos

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Live zebrafish embryos (54 hpf – 8 dpf) were anesthetized
with 0.01% tricaine (Fisher), then mounted in 4%–6% (wt/vol)
methylcellulose in 35-mm imaging dishes (MatTek) as described previously
(Renaud et al., 2011 (link)).
Fluorescent imaging of
Tg(drl:creERT2;ubi:Switch),
Tg(drl:CFP-NTR;drl:mCherry),
Tg(drl:CFP-NTR;cd41:GFP),
Tg(drl:CFP-NTR;lyz:dsRed), and
Tg(drl:CFP-NTR;rag2:mCherry) were performed with Zeiss
Discovery.V8 and Zeiss Axio Observer A1 Inverted microscope with an AxioCam
HRc Zeiss camera and Zeiss Zen 2 or 2.5 software. Fluorescence was detected
with cyan fluorescent protein (CFP), mCherry, Texas Red (for dsRed lines),
and green fluorescent protein (GFP) filters.
A Zeiss Live DuoScan confocal microscope with AIM 4.2 Software was
used to visualize co-expression in
Tg(drl:CFP-NTR+;runx1:mCherry+)embryos using 405nm and 561nm excitation wavelength. Embryos (6 dpf) were
anesthetized with 0.01% tricaine (Fisher), oriented in a drop of 3% wt/vol
methylcellulose, then mounted in 1% agarose in 35-mm imaging dishes (MatTek)
as described previously (Renaud et al.,
2011).
Ulloa B.A., Habbsa S.S., Potts K.S., Lewis A., McKinstry M., Payne S.G., Flores J.C., Nizhnik A., Norberto M.F., Mosimann C, & Bowman T.V. (2021). Definitive hematopoietic stem cells minimally contribute to embryonic hematopoiesis. Cell reports, 36(11), 109703.
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2

Fluorescent Imaging of Zebrafish Embryos

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Live zebrafish embryos (54 hpf – 8 dpf) were anesthetized
with 0.01% tricaine (Fisher), then mounted in 4%–6% (wt/vol)
methylcellulose in 35-mm imaging dishes (MatTek) as described previously
(Renaud et al., 2011 (link)).
Fluorescent imaging of
Tg(drl:creERT2;ubi:Switch),
Tg(drl:CFP-NTR;drl:mCherry),
Tg(drl:CFP-NTR;cd41:GFP),
Tg(drl:CFP-NTR;lyz:dsRed), and
Tg(drl:CFP-NTR;rag2:mCherry) were performed with Zeiss
Discovery.V8 and Zeiss Axio Observer A1 Inverted microscope with an AxioCam
HRc Zeiss camera and Zeiss Zen 2 or 2.5 software. Fluorescence was detected
with cyan fluorescent protein (CFP), mCherry, Texas Red (for dsRed lines),
and green fluorescent protein (GFP) filters.
A Zeiss Live DuoScan confocal microscope with AIM 4.2 Software was
used to visualize co-expression in
Tg(drl:CFP-NTR+;runx1:mCherry+)embryos using 405nm and 561nm excitation wavelength. Embryos (6 dpf) were
anesthetized with 0.01% tricaine (Fisher), oriented in a drop of 3% wt/vol
methylcellulose, then mounted in 1% agarose in 35-mm imaging dishes (MatTek)
as described previously (Renaud et al.,
2011).
Ulloa B.A., Habbsa S.S., Potts K.S., Lewis A., McKinstry M., Payne S.G., Flores J.C., Nizhnik A., Norberto M.F., Mosimann C, & Bowman T.V. (2021). Definitive hematopoietic stem cells minimally contribute to embryonic hematopoiesis. Cell reports, 36(11), 109703.
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3

Confocal Imaging of Vasculature Development

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For confocal images, the embryos were immobilized and embedded in 1.5% low-melting-point agarose with 5% tricaine (Invitrogen), and images were obtained on the Zeiss LSM700 or Nikon Eclipse 90i C1 confocal microscopes and processed using the ImageJ software (NIH, USA). The counting area was between the 5th and 15th ISVs of the 24–72-hpf embryos. The number of cells in the ISVs was determined by counting cells in the individual slices of confocal stacks. Final figures were processed using Adobe Photoshop.
Wu T.Y., Wang Y.S., Song Y.C., Chen Z.Y., Chen Y.T., Chiu C.C, & Wu C.Y. (2017). Fine-tune regulation of carboxypeptidase N1 controls vascular patterning during zebrafish development. Scientific Reports, 7, 1852.
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4

Embryo Mounting and Imaging

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Embryos were mounted either in 3% methyl cellulose (Sigma) or 1.5% low melt agarose (Invitrogen, Philadelphia, PA, USA). Images were photographed with an AxioCam high-resolution camera and processed by AxioVision software (Carl Zeiss, Jena, Germany). For the confocal images, embryos were immobilized and embedded in 1.5% low-melting-point agarose with 5% tricaine (Invitrogen), and images were collected on a Zeiss LSM700 or Nikon Eclipse 90i C1 confocal microscope and processed with ImageJ software (NIH, Bethesda, MD, USA). Final figures were made using Adobe Photoshop.
Chang H.W., Wang W.D., Chiu C.C., Chen C.H., Wang Y.S., Chen Z.Y., Liu W., Tai M.H., Wen Z.H, & Wu C.Y. (2017). Ftr82 Is Critical for Vascular Patterning during Zebrafish Development. International Journal of Molecular Sciences, 18(1), 156.
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5

Zebrafish Pronephros Dextran Uptake

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Lysine-fixable 10 kDa dextran (Alexa Fluor 488 conjugated) or 500 kDa dextran [fluorescein isothiocyanate (FITC) conjugated] (Thermo Fisher Scientific) were prepared in PBS at 2 µg/µl final concentration. In addition, recombinant Cy3-conjugated His-tagged RAP (39 kDa), prepared in PBS at 5 µg/µl final concentration, was kindly provided by Dr Martin Lowe (University of Manchester, Manchester, UK). Zebrafish embryos were anesthetized in tricaine (0.013% w/v; Fisher Scientific) diluted in embryo water at 72 hpf. Approximately 0.5-1 nl of dextran or RAP was injected into the common cardinal vein using a glass micropipette and a pneumatic pressure injector (PLI90; Harvard Apparatus) and micromanipulator. Uptake in the renal tubular cells of the proximal pronephros was analyzed at 1-2.5 hpi, using a Nikon SMZ18 fluorescent stereomicroscope with an image capture system. High dextran uptake was defined as >20 fluorescent puncta observed along the proximal pronephros. Low dextran uptake was defined as one to 20 fluorescent puncta observed along the proximal pronephros, and no uptake indicated that no fluorescent puncta were seen. Animals injected with 500 kDa dextran were analyzed at 24 hpi.
Ates K.M., Wang T., Moreland T., Veeranan-Karmegam R., Ma M., Jeter C., Anand P., Wenzel W., Kim H.G., Wolfe L.A., Stephen J., Adams D.R., Markello T., Tifft C.J., Settlage R., Gahl W.A., Gonsalvez G.B., Malicdan M.C., Flanagan-Steet H, & Pan Y.A. (2020). Deficiency in the endocytic adaptor proteins PHETA1/2 impairs renal and craniofacial development. Disease Models & Mechanisms, 13(5), dmm041913.
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6

Microscopic Imaging of Cyclopic Embryos

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Using brightfield light microscopy, 10-20 single injected embryos were examined and embryos showing cyclopia were imaged using a Zeiss Lumar Lamar V12 stereo microscope and AxioVision SE64 Rel.4.9.1 software (Zeiss, Oberkochen, Germany). Embryos were anaesthetized in a solution of embryo medium containing 2 mM tricaine (Sigma-Aldrich, St.
Louis, MO) to prevent gross movements during imaging. Embryos were mounted in 3 % methyl cellulose solution (Fisher Scientific, Hampton, NH) to orient them and were imaged in the continued presence of 2 mM tricaine.
Turner A.N., Andersen R.S., Bookout I.E., Brashear L.N., Davis J.C., Gahan D.M., Davis J.C., Gotham J.P., Hijaz B.A., Kaushik A.S., Mcgill J.B., Miller V.L., Moseley Z.P., Nowell C.L., Patel R.K., Rodgers M.C., Patel R.K., Shihab Y.A., Walker A.P., Glover S.R., Foster S.D, & Challa A.K. (2018). Analysis of novel domain-specific mutations in the zebrafish ndr2/cyclops gene generated using CRISPR-Cas9 RNPs. Journal of genetics, 97(5).
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7

Visualizing Spermathecae in Live Hermaphrodite Worms

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To image spermathecae in live animals, adult hermaphrodite worms were
paralyzed in M9 containing 0.1% tricaine (Acros Organics, Geel, Belgium) and
0.01% levamisole (Acros Organics, Geel, Belgium), and live mounted onto a 2%
agarose pad. Epifluorescence and differential interference contrast (DIC) images
were obtained using an Eclipse 90i research upright microscope (Nikon, Tokyo,
Japan) at room temperature using a CFI Plan Apochromat 403/NA 1.0 oil immersion
objective, with a Cool-SNAP HA2 digital monochrome charge-coupled device camera
(Photometrics, Tucson, AZ) driven by NIS-Elements AR acquisition and analysis
software (version 3.1; Nikon). Ovulation movies were acquired using DIC
microscopy as previously described (Hegsted et
al., 2016). Spermathecae expressing mCherry were scored for
localization of GFP as junctional, nuclear, or cytoplasmic. For spermathecae
selected for imaging, Z-stacks were collected at 2.5 μm intervals before
deconvolution in NIS-Elements using AutoQuant Blind Deconvolution, with 17
iterations and medium noise level. Spermathecae outlines indicted in Figure 5 B were determined using mCherry
fluorescence.
Hegsted A., Votra S., Christophe A.M., Yingling C.V., Sundaramurthy S, & Pruyne D. (2019). Functional importance of an inverted formin C-terminal tail at morphologically dynamic epithelial junctions. Cytoskeleton (Hoboken, N.J.), 76(4), 322-336.
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8

Zebrafish Heart Rate Monitoring

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Zebrafish express Zerg, an orthologue of hERG, and many hERG inhibitors induce bradycardia an arrhythmia in zebrafish.35 (link) Heart rate was recorded and calculated as reported previously36 (link) with slight modifications (n = 3–9). Briefly, 7 dpf zebrafish larvae were anesthetized with tricaine (Acros Organics) and immobilized in a lateral orientation using 1% low melt agarose (LMA, Gene Mate) dissolved in egg water.37 tricaine was washed out and drug was added to 4 mL embryo media in a 6-well plate (final concentration = 50 μM). Videos were collected at 30 frames per second (fps) using a Leica M80 scope with an ACHRO 1x nosepiece attachment and a Leica IC80 HD camera. Regions of interest (ROIs) were drawn around the atrium and ventricle of individual zebrafish and average pixel dynamics were calculated using the ImageJ plugin Time Series Analyzer V3. This pixel change oscillation was graphically smoothed using the Savgol filter in SciPy. Peaks were detected using the SciPy package “find_peaks”. Peak time interval and BPM were calculated using custom code. The arrythmia score was calculated as the ratio of atrium BPM to ventricle BPM (n = 6–18).
Cameron L.P., Tombari R.J., Lu J., Pell A.J., Hurley Z.Q., Ehinger Y., Vargas M.V., McCarroll M.N., Taylor J.C., Myers-Turnbull D., Liu T., Yaghoobi B., Laskowski L.J., Anderson E.I., Zhang G., Viswanathan J., Brown B.M., Tjia M., Dunlap L.E., Rabow Z.T., Fiehn O., Wulff H., McCorvy J.D., Lein P.J., Kokel D., Ron D., Peters J., Zuo Y, & Olson D.E. (2020). A Non-Hallucinogenic Psychedelic Analog with Therapeutic Potential. Nature, 589(7842), 474-479.
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9

Zebrafish Heart Rate Monitoring

Check if the same lab product or an alternative is used in the 5 most similar protocols
Zebrafish express Zerg, an orthologue of hERG, and many hERG inhibitors induce bradycardia an arrhythmia in zebrafish.35 (link) Heart rate was recorded and calculated as reported previously36 (link) with slight modifications (n = 3–9). Briefly, 7 dpf zebrafish larvae were anesthetized with tricaine (Acros Organics) and immobilized in a lateral orientation using 1% low melt agarose (LMA, Gene Mate) dissolved in egg water.37 tricaine was washed out and drug was added to 4 mL embryo media in a 6-well plate (final concentration = 50 μM). Videos were collected at 30 frames per second (fps) using a Leica M80 scope with an ACHRO 1x nosepiece attachment and a Leica IC80 HD camera. Regions of interest (ROIs) were drawn around the atrium and ventricle of individual zebrafish and average pixel dynamics were calculated using the ImageJ plugin Time Series Analyzer V3. This pixel change oscillation was graphically smoothed using the Savgol filter in SciPy. Peaks were detected using the SciPy package “find_peaks”. Peak time interval and BPM were calculated using custom code. The arrythmia score was calculated as the ratio of atrium BPM to ventricle BPM (n = 6–18).
Cameron L.P., Tombari R.J., Lu J., Pell A.J., Hurley Z.Q., Ehinger Y., Vargas M.V., McCarroll M.N., Taylor J.C., Myers-Turnbull D., Liu T., Yaghoobi B., Laskowski L.J., Anderson E.I., Zhang G., Viswanathan J., Brown B.M., Tjia M., Dunlap L.E., Rabow Z.T., Fiehn O., Wulff H., McCorvy J.D., Lein P.J., Kokel D., Ron D., Peters J., Zuo Y, & Olson D.E. (2020). A Non-Hallucinogenic Psychedelic Analog with Therapeutic Potential. Nature, 589(7842), 474-479.
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