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Fast green fcf dye

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Fast Green FCF is a synthetic green dye used as a colorant in various laboratory applications. It is a water-soluble dye that can be used for staining and visualization purposes in various scientific and industrial processes. The dye has a characteristic green color and is commonly used in biochemical and analytical techniques.

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Lab products found in correlation

Nanoject injector, by Drummond (3 mentions) Aav gfp, by Addgene (2 mentions) Fast green fcf dye, by Drummond (2 mentions) Aav flex tdtomato, by Addgene (2 mentions) Syringe pump, by Harvard Apparatus (2 mentions) Nanoject 2 injector, by Drummond (2 mentions) Swiss webster, by Taconic Biosciences (1 mentions) 32 gauge needle, by Hamilton Company (1 mentions) 5 μl syringe, by Hamilton Company (1 mentions) Vicryl suture, by Johnson & Johnson (1 mentions) Cuy650p5, by Nepa Gene (1 mentions) Cuy21sc, by Nepa Gene (1 mentions) Biorad xcell electroporation cuvettes, by Bio-Rad (1 mentions) Nuclepore track etch membrane filters, by Cytiva (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Nanoject 3 injector, by Drummond (1 mentions) Fmr1 y, by Jackson ImmunoResearch (1 mentions) X linked gfp mice, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

Fast Green FCF Fast Green dye Fast Green

12 protocols using fast green fcf dye

1

Viral Vector Injections in Rodents

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For all injections Fast Green FCF dye (MilliporeSigma) was included to aid visualization.
For IP AAV9 injections, ~1×1012 vg was delivered through glass pipettes into P0-P1 animals after the animals were anesthetized with ice.
For all parenchymal AAV1 injections, viruses were diluted to final concentrations of 3×1012-1×1013 vg/mL each before injection. 3-4 injections were made on different sites and 50-100 nL was injected at each site. For non-stereotactic cortical AAV1 injections, P1-P3 animals were anesthetized with ice, and viruses were injected into the cortex using glass pipettes directly through the skulls. For stereotactic cortical AAV1 injections, P21 animals were anesthetized with isoflurane, and viruses were injected into S1 forelimb area using glass pipettes through drilled holes in the skull. For spinal cord AAV1 injections, P12-P14 animals were anesthetized with isoflurane, and viruses were injected into the cervical spinal cord using glass pipettes directly through the meninges.
Zhang Q., Lee W.C., Paul D.L, & Ginty D.D. (2019). Multiplexed peroxidase-based electron microscopy labeling enables simultaneous visualization of multiple cell types. Nature neuroscience, 22(5), 828-839.
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2

In Utero Electroporation of Mouse Embryos

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Embryonic day (E) 15.5 timed-pregnant female Swiss Webster (Taconic Biosciences) mice were deeply anaesthetized with 2% isoflurane. Uterine horns were exposed and periodically rinsed with warm sterile 1X PBS. Plasmid DNA mixture was injected into the lateral ventricle of one cerebral hemisphere of an embryo using a 32-gauge needle (Hamilton Company) attached to a 5 μl syringe (Hamilton Company). Final plasmid DNA concentration was 4.5 μg/μl in water (DNA mass ratio of pAAV-CAG-S1-GCaMP6f, pAAV-CAG-S3-ExRaiAKAR, and pAAV-CAG-mRuby3–6xFLAG at 1:2:2). Fast Green FCF dye (Millipore Sigma) was added to the DNA mixture to visualize the mixture during injection. Five voltage pulses (50 V, 50 ms duration, 1 Hz) were delivered two times using 5-mm round plate electrodes (Harvard Apparatus), with the cathode placed on top of the skull to target the hippocampus. Electroporated embryos were placed back into the dam, and allowed to mature for delivery.
Linghu C., Johnson S.L., Valdes P.A., Shemesh O.A., Park W.M., Park D., Piatkevich K.D., Wassie A.T., Liu Y., An B., Barnes S.A., Celiker O.T., Yao C.C., Yu C.C., Wang R., Adamala K.P., Bear M.F., Keating A.E, & Boyden E.S. (2020). Spatial multiplexing of fluorescent reporters for imaging signaling network dynamics. Cell, 183(6), 1682-1698.e24.
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3

Viral Vector Injections in Rodents

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For all injections Fast Green FCF dye (MilliporeSigma) was included to aid visualization.
For IP AAV9 injections, ~1×1012 vg was delivered through glass pipettes into P0-P1 animals after the animals were anesthetized with ice.
For all parenchymal AAV1 injections, viruses were diluted to final concentrations of 3×1012-1×1013 vg/mL each before injection. 3-4 injections were made on different sites and 50-100 nL was injected at each site. For non-stereotactic cortical AAV1 injections, P1-P3 animals were anesthetized with ice, and viruses were injected into the cortex using glass pipettes directly through the skulls. For stereotactic cortical AAV1 injections, P21 animals were anesthetized with isoflurane, and viruses were injected into S1 forelimb area using glass pipettes through drilled holes in the skull. For spinal cord AAV1 injections, P12-P14 animals were anesthetized with isoflurane, and viruses were injected into the cervical spinal cord using glass pipettes directly through the meninges.
Zhang Q., Lee W.C., Paul D.L, & Ginty D.D. (2019). Multiplexed peroxidase-based electron microscopy labeling enables simultaneous visualization of multiple cell types. Nature neuroscience, 22(5), 828-839.
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4

Viral Vector Delivery to NJP Ganglia

Cited 1 time
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NJP ganglia were injected with AAVs or diphtheria toxin as previously described34 (link), with minor modifications. Mice were anaesthetized (200 mg kg−1 avertin, intraperitoneal injection), and depth of anaesthesia ensured throughout the procedure by lack of a toe pinch response. NJP ganglia were exposed and serially injected (10 × 13.8 nl) with saline solution containing either AAVs (titre > 6.7 × 1012 vg ml−1) or diphtheria toxin (5 μg ml−1, Sigma) supplemented with 0.05% Fast Green FCF Dye (Sigma) using a sharply pulled glass pipette attached to a Nanoject Injector (Drummond). AAV-cre (AAV-Syn-Cre-GFP, SignaGen Laboratories, SL100892, AAV9), AAV-flex-tdTomato (AAV9.CAG.Flex.tdTomato.WPRE.BGH, Addgene, 51503, AAV9), AAV-GFP (pENN.AAV.CB7.CI.eGFP.WPRE.rBG, Addgene, 105542, AAV9), and AAV-flex-AP (AAV9.CAG.flex.PLAP.WPRE.bgH, custom virus, Boston Children’s Hospital Viral Core, Boston, MA, AAV9) were purchased. Successful injection was verified by Fast Green FCF Dye slowly filling the entire ganglion without leakage. Surgical wounds were closed with coated VICRYL Sutures (J392H, Ethicon) and animals received Buprenorphine SR (BupSR-LAB, ZooPharm, 20 mg kg−1 subcutaneously at the back of the neck) as an analgesic. Animals recovered for at least two weeks for behavioural analysis or four weeks for histological analysis.
Bin N.R., Prescott S.L., Horio N., Wang Y., Chiu I.M, & Liberles S.D. (2023). An airway-to-brain sensory pathway mediates influenza-induced sickness. Nature, 615(7953), 660-667.
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5

In Utero Electroporation of Embryonic Neocortex

Cited 3 times
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IUE was performed as described previously (Usui et al. 2017 (link)). Timed pregnant mice were deeply anesthetized with isoflurane (1.5%–3% in oxygen) during surgery and were also administered 1 mg/kg buprenorphine to limit pain after electroporation. Endotoxin-free plasmid DNA (1–2 µg/µL) with 0.1% Fast Green FCF dye (Sigma-Aldrich, F7252) was microinjected into the lateral ventricles of E14.5 embryos to target upper layer (layer 2/3) neurons of the neocortex. The embryo was held through the uterus with platinum plate electrode tweezers (Protech International, Inc., CUY650P5) and electroporated (five 50-msec pulses of 33 V with an interval of 950 msec) (NEPA Gene, CUY21SC) (Baek et al. 2015 (link); Li et al. 2015 (link)). Electroporated embryos were analyzed at E18.5.
Usui N., Araujo D.J., Kulkarni A., Co M., Ellegood J., Harper M., Toriumi K., Lerch J.P, & Konopka G. (2017). Foxp1 regulation of neonatal vocalizations via cortical development. Genes & Development, 31(20), 2039-2055.
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6

Embryonic Lung Kaleidoscope Imaging

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Time-mated females were harvested at E14.5 and lungs with trachea and glottis were removed from embryos. The Cre-recombined ROSA-Kaleidoscope plasmid and Fast Green FCF dye (MKCD1540; Sigma-Aldrich) were mouth-pipetted through the glottis until embryonic lumens were visibly filled. Lungs were electroporated in BioRad Xcell electroporation cuvettes (165-2088; BioRad) at 35 V for three rounds of 25 ms with 1-s intervals. Lungs were then placed on Nuclepore Track-Etch Membrane filters (10417101; Whatman) and floated on RPMI-1640 media with 10% FBS 1% PenStrep for 24 h before fixation in 0.5% PFA for 4 h. After PBS wash, lungs were embedded in optimal cutting temperature compound (OCT; 4583; Tissue-Tek), cryosectioned, and imaged on Deltavision Deconvolution scope with 60× oil objective (Plan-Apo NA 1.42) running SoftWoRx.
Hutchison V., Lynch A., Gutierrez-Gamez A.M, & Chen J. (2023). Inducible tricolor reporter mouse for parallel imaging of lysosomes, mitochondria, and microtubules. The Journal of Cell Biology, 223(1), e202305086.
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7

AAV Transduction of Nodose Ganglia

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AAV-flex-tdTomato (Addgene, 51502-AAV9), AAV-Gfp (Addgene, 105542-AAV9), and AAV-mCherry (Addgene, 105544-AAV9) were purchased. Surgically exposed NJP superganglia were serially injected (10 × 13.8 nl) with AAV injection solution (AAV titer > 6.7 × 1012 vg/ml and 0.05% Fast Green FCF Dye, Sigma) or DT injection solution (5 μg/ml DT Sigma D0564, 0.05% Fast Green FCF Dye, PBS) using a Nanoject Injector (Drummond). Dye typically filled the NJP ganglion, but occasionally, the injection needle was repositioned for maximal dye spread. In control experiments, ectopic AAV infection of superior cervical ganglia was not observed by fluorescence microscopy. After AAV infection, animals were sacrificed four weeks later for histological analysis. After DT injection, animals were used at least two weeks later for physiological analysis, and the extent of ablation was analyzed post hoc by DTR immunostaining of NJP ganglia.
Min S., Chang R.B., Prescott S.L., Beeler B., Joshi N.R., Strochlic D.E, & Liberles S.D. (2019). Arterial Baroreceptors Sense Blood Pressure through Decorated Aortic Claws. Cell reports, 29(8), 2192-2201.e3.
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8

In Utero Microinjection of Embryonic Mouse Otocysts

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All animal surgeries were conducted in a sterile environment in the laboratory of the Animal Center at National Yang-Ming University. The detailed surgical procedures used in this study were adopted from procedures described previously.38 Briefly, pregnant C57BL/6 mice were anesthetized with gaseous 0.4% isoflurane (Panion & BF Biotech, Taipei, Taiwan) in a sealed cage for 3–5 min until they were deeply anesthetized. The mice were then removed from the cage and fitted with masks for continuous administration of gaseous isoflurane before the abdominal skin was incised along the midline for 1.5 cm. One otocyst in each embryo was microinjected by glass micropipette. Each otocyst in the experimental group was microinjected with a mixture of 1 μL of AAV2/Anc80L65-CMV-eGFP and Fast Green FCF dye (Sigma-Aldrich, St. Louis, MO, USA; 5:1 ratio), while each otocyst in the sham group was microinjected with 1 μL of phosphate-buffered saline (PBS) (Thermo Fisher Scientific, Waltham, MA, USA; Figure 1A). During surgery, we regularly dripped 50°C PBS onto the surface of the uterus to prevent heat loss and to allow the syringe needle to penetrate easily. After microinjection, we sequentially closed the abdominal wound in the muscular and dermal layers with surgical sutures. Mice were placed under a lamp for 5 min after surgery to recover.
Hu C.J., Lu Y.C., Tsai Y.H., Cheng H.Y., Takeda H., Huang C.Y., Xiao R., Hsu C.J., Tsai J.W., Vandenberghe L.H., Wu C.C, & Cheng Y.F. (2020). Efficient in Utero Gene Transfer to the Mammalian Inner Ears by the Synthetic Adeno-Associated Viral Vector Anc80L65. Molecular Therapy. Methods & Clinical Development, 18, 493-500.
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9

AAV Transduction of Nodose Ganglia

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AAV-flex-tdTomato (Addgene, 51502-AAV9), AAV-Gfp (Addgene, 105542-AAV9), and AAV-mCherry (Addgene, 105544-AAV9) were purchased. Surgically exposed NJP superganglia were serially injected (10 × 13.8 nl) with AAV injection solution (AAV titer > 6.7 × 1012 vg/ml and 0.05% Fast Green FCF Dye, Sigma) or DT injection solution (5 μg/ml DT Sigma D0564, 0.05% Fast Green FCF Dye, PBS) using a Nanoject Injector (Drummond). Dye typically filled the NJP ganglion, but occasionally, the injection needle was repositioned for maximal dye spread. In control experiments, ectopic AAV infection of superior cervical ganglia was not observed by fluorescence microscopy. After AAV infection, animals were sacrificed four weeks later for histological analysis. After DT injection, animals were used at least two weeks later for physiological analysis, and the extent of ablation was analyzed post hoc by DTR immunostaining of NJP ganglia.
Min S., Chang R.B., Prescott S.L., Beeler B., Joshi N.R., Strochlic D.E, & Liberles S.D. (2019). Arterial Baroreceptors Sense Blood Pressure through Decorated Aortic Claws. Cell reports, 29(8), 2192-2201.e3.
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10

Tracing Neuronal Projections via AAV

Cited 1 time
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AAV injections into NJP ganglia were done as reported previously (Chang et al., 2015 (link)), with minor modifications. Surgically exposed NJP superganglia were serially injected (10 × 10 nl) with AAV solutions containing 0.05% Fast Green FCF Dye (Sigma) using a Nanoject III Injector (Drummond). AAV-flex-tdTomato was AAV9.CAG.Flex.tdTomato.WPRE.BGH (Addgene viral prep #51503-AAV9, ~1013 genome copies/ml). AAV-flex-AP was AAV9.CAG.flex.PLAP.WPRE.BGh (custom virus, Boston Children’s Hospital Viral Core, ~1013 genome copies/ml). AAV-flex-tdTomato virus solution was diluted 1:1 with a Cre-independent AAV-GFP virus solution (pENN.AAV.CB7.CI.eGFP.WPRE.rBG, Addgene viral prep #105542-AAV9, ~1013 genome copies/ml) prior to injection. Animals recovered from surgery and were sacrificed 4 weeks later for histology.
Prescott S.L., Umans B.D., Williams E.K., Brust R.D, & Liberles S.D. (2020). An airway protection program revealed by sweeping genetic control of vagal afferents. Cell, 181(3), 574-589.e14.
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