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Lsm700

Manufactured by Nikon
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The LSM700 is a compact and versatile laser scanning microscope designed for a wide range of applications. It features a high-resolution image acquisition system and advanced optics to provide detailed analysis of samples. The core function of the LSM700 is to enable high-quality imaging and visualization of microscopic specimens.

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Lab products found in correlation

Eclipse 90i c1, by Nikon (4 mentions) Low melting point agarose, by Thermo Fisher Scientific (3 mentions) Methylcellulose, by Merck Group (3 mentions) Axiocam hrc camera, by Zeiss (2 mentions) Tricaine, by Thermo Fisher Scientific (2 mentions) Image pro plus software 7, by Media Cybernetics (2 mentions) Zen 2009 light edition software, by Zeiss (2 mentions) Nis element 5, by Nikon (2 mentions) Axioskope 2 microscope, by Zeiss (2 mentions) Axiocammr3 camera, by Zeiss (2 mentions) A1 confocal microscope, by Nikon (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Zen software, by Zeiss (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) A1r confocal microscope, by Nikon (1 mentions) Low melt agarose, by Thermo Fisher Scientific (1 mentions) Axiocam high, by Zeiss (1 mentions) Axiovision software, by Zeiss (1 mentions) Eclipse ti2 e, by Nikon (1 mentions) Sliding microtome, by Leica camera (1 mentions)

10 protocols using lsm700

1

Imaging Techniques for Embryo Visualization

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Embryos were mounted in 3% methylcellulose (Sigma) or 1.5% low melting point agarose (Invitrogen) and image captured with a color digital AxioCam HRc camera (Carl Zeiss, Jena, Germany) or SPOT RT3 camera (Diagnostic Inc., Sterling heights, MI, USA). For the confocal images, embryos were embedded with 5% tricaine and images were collected on a Zeiss LSM700 or Nikon Eclipse 90i C1 confocal microscopes, and Z-stacked images (generally 30 slices with 5–10 μm between) were processed with ZEN software (Carl Zeiss) or ImageJ software (NIH, Bethesda, MD, USA).
Song Y.C., Wu B.J., Chiu C.C., Chen C.L., Zhou J.Q., Liang S.R., Duh C.Y., Sung P.J., Wen Z.H, & Wu C.Y. (2017). Coral-Derived Natural Marine Compound GB9 Impairs Vascular Development in Zebrafish. International Journal of Molecular Sciences, 18(8), 1696.
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2

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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3

Multiparametric Imaging of Atherosclerotic Lesions

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Movat, IL1R1, MMP3, Ter119 and Sudan IV staining of brachiocephalic arteries were imaged using a Zeiss Axioskope2 microscope equipped with an AxioCamMR3 camera. Image acquisition was performed with AxioVision40 version 4.6.3.0 software (Carl Zeiss Imaging Solution). Digitized images were analyzed with Image Pro Plus Software 7.0 (Media Cybernetics). Immunofluorescent staining was imaged using either a Zeiss LSM700 or a Nikon A1 confocal microscope to acquire a series of eight z-stack images at 1-μm intervals. Zen 2009 Light Edition Software (Zeiss) or NIS-Element 5.02 Software (Nikon) were used for analysis of each z-stack image and single-cell counting was performed for phenotyping and quantifying the cell population comprised within the 30μm thick layer proximal to the lumen (i.e., fibrous cap area). Assessment of YFP+, ACTA2+, and LGALS3+ areas (normalized to lesion or fibrous cap area) was performed using maximal intensity projection images and the analysis was done using Image Pro Plus Software 7.0 (Media Cybernetics). Maximal intensity projection of representative images were used to generate the representative images included in the figures and Adobe Photoshop was used to process and format images.
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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4

3D Culture of Breast Cancer Cell Lines

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MCF-10A cell were obtained from Dr. Israel Vlodavsky (Technion Ins, Haifa, Israel) and were maintained as described previously.48 (link) MCF-7 and T47D cells were obtained from American Type Culture Collection (ATCC) and were stably transected with pCDNA3-Intβ3 (a gift from Dr. Newman PJ; The Blood Research Center of Southeastern Wisconsin Inc.). Clones of MCF-7-Intβ3 and pool of T47D-Intβ3 were selected by G418 (Gold Biotechnology, St. Louis, MO, USA) and were maintained in DMEM or RPMI supplemented with 10% fetal bovine serum and antibiotics (Life Technologies, Hertzliya Pituach, Israel), respectively. 3D cultures were carried out in growth factor-reduced Cultrex Basement Membrane Extract (Trevigen, Inc., Gaithersburg, MD, USA) 49 (link) as previously described.48 (link), 50 Cilengitide, was a kind gift from Dr. Ronit Satchi Fainaro (University of Tel Aviv). Immunofluorescent images were captured by either Zeiss LSM 700 or by Nikon A1R confocal microscope.
Abu-Tayeh H., Weidenfeld K., Zhilin-Roth A., Schif-Zuck S., Thaler S., Cotarelo C., Tan T.Z., Thiery J.P., Green J.E., Klorin G., Sabo E., Sleeman J.P., Tzukerman M, & Barkan D. (2016). ‘Normalizing' the malignant phenotype of luminal breast cancer cells via alpha(v)beta(3)-integrin. Cell Death & Disease, 7(12), e2491-.
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5

Multiparametric Imaging of Atherosclerotic Lesions

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Movat, IL1R1, MMP3, Ter119 and Sudan IV staining of brachiocephalic arteries were imaged using a Zeiss Axioskope2 microscope equipped with an AxioCamMR3 camera. Image acquisition was performed with AxioVision40 version 4.6.3.0 software (Carl Zeiss Imaging Solution). Digitized images were analyzed with Image Pro Plus Software 7.0 (Media Cybernetics). Immunofluorescent staining was imaged using either a Zeiss LSM700 or a Nikon A1 confocal microscope to acquire a series of eight z-stack images at 1-μm intervals. Zen 2009 Light Edition Software (Zeiss) or NIS-Element 5.02 Software (Nikon) were used for analysis of each z-stack image and single-cell counting was performed for phenotyping and quantifying the cell population comprised within the 30μm thick layer proximal to the lumen (i.e., fibrous cap area). Assessment of YFP+, ACTA2+, and LGALS3+ areas (normalized to lesion or fibrous cap area) was performed using maximal intensity projection images and the analysis was done using Image Pro Plus Software 7.0 (Media Cybernetics). Maximal intensity projection of representative images were used to generate the representative images included in the figures and Adobe Photoshop was used to process and format images.
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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6

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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7

Confocal Imaging of Mitotic Cells

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Three-dimensional image stacks of mitotic cells were acquired under a laser-scanning confocal microscope (Carl Zeiss; LSM700) or an inverted microscope (Nikon Corporation; ECLIPSE Ti2-E). The z-stacks were scanned with a step size of 0.38 µm or 0.2 µm.
Ryu J, & Kim J.E. (2022). CCAR2 controls mitotic progression through spatiotemporal regulation of Aurora B. Cell Death & Disease, 13(6), 534.
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8

Immunohistochemical Analysis of Mouse Brain

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Mice were euthanized and perfused with intracardial injection of 4% (w/v) paraformaldehyde in phosphate-buffered saline (PBS). Brains were isolated and post-fixed overnight with 4% (w/v) paraformaldehyde at 4 °C. After cryoprotection with 30% sucrose in PBS for 72 h at 4 °C, 40-μm brain slices were collected through a sliding microtome (Leica). For immunostaining, brain sections were treated with 50% formamide in 1 X SSC buffer for 2 h at 65 °C. For BrdU staining, the sections were pretreated with 2N HCl for 30 min at 37 °C. The following primary antibodies were used: GFP (Chicken; 1:2,000; Aves Labs), mCherry (Goat; 1:2,000; Mybiosource), NeuN (Rabbit; 1:10,000; Abcam), GFAP (Mouse; 1:2,000; Sigma), Cre (Rabbit; 1:500; Covance), ALDH1L1 (Mouse; 1:200; Neuromab), ALDOC (Goat; 1:100; Santa Cruz), BrdU (Rat; 1:500; Bio-Rad), NEUROD1 (Rabbit; 1:1,000; Abcam), PTBP1 (Rabbit, 1:1,000, ABclonal), FLAG (Rabbit; 1:300; Sigma), HA (Chicken; 1:300; Aves), DCX (Goat; 1:500; Santa Cruz), and MAP2 (Mouse; 1:1,000; Proteintech). Alexa Fluor 488-, 555-, or 647-conjugated corresponding secondary antibodies from Jackson ImmunoResearch were used for indirect fluorescence (1:2,000). Nuclei were counterstained with Hoechst 33342 (Hst). Images were captured using a Zeiss LSM700 or Nikon A1R confocal microscope for analysis.
Wang L.L., Serrano C., Zhong X., Ma S., Zou Y, & Zhang C.L. (2021). Revisiting astrocyte to neuron conversion with lineage tracing in vivo. Cell, 184(21), 5465-5481.e16.
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9

Imaging Embryos with Agarose or Methylcellulose

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Embryos were mounted with either 1.5% low-melting-point agarose (Invitrogen) or 3% methylcellulose (Sigma), and the images were acquired with a color digital AxioCam HRc camera (Carl Zeiss, Jena, Germany) or a SPOT RT3 camera (Diagnostic Inc., Sterling heights, MI, USA). For confocal imaging, embryos were embedded with 5% tricaine, and images were acquired with a Zeiss LSM700 or Nikon Eclipse 90i C1 confocal microscope and processed with ImageJ software (NIH, Bethesda, MD, USA).
Chiu C.C., Chin H.K., Chung S.Y., Hsieh K.H., Huang Y.S., Huang M.F., Lo Y.H., Wen Z.H, & Wu C.Y. (2022). Nitrobenzoate-Derived Compound X8 Impairs Vascular Development in Zebrafish. International Journal of Molecular Sciences, 23(14), 7788.
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10

Whole Mount Zebrafish Antibody Staining

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Antibody staining was performed as described [22 (link)]. Briefly, whole mount zebrafish samples were stained with primary antibodies: mouse anti-TNFα (1:50; Abcam #52B83, Cambridge, UK), guinea pig anti-insulin (1:200, Invitrogen #180067, Carlsbad, CA, USA), rabbit anti-cleaved caspase 3 (1:100; Cell Signaling Technologies #9661S, Danvers, MA, USA), mouse anti-Prox1 (1:50, Developmental Studies Hybridoma Bank, AB_2619013, Iowa City, IA, USA), and chicken anti-GFP (1:200, Aves labs #GFP-1010, Davis, CA, USA). Primary antibodies were detected with complementary Alexa-conjugated secondary antibodies (1:500, Jackson ImmunoResearch, West Grove, PA, USA). DNA was stained with TO-PRO3 (1:500, Thermo Fisher #T3605, Waltham, MA, USA) or DAPI (1:400, Thermo Fisher #62248, Waltham, MA, USA). After staining, larvae were mounted on slides in VECTASHIELD (Vector Labs H-1000). Confocal imaging was performed with a Zeiss LSM700 or Nikon A1 microscope. Images of livers were analyzed using NIH Fiji software. Images following immunofluorescence staining were used to verify the presence of TNFα and H2BGFP in livers and to count infiltrating macrophages.
Kulkarni A., Ibrahim S., Haider I., Basha A., Montgomery E., Ermis E., Mirmira R.G, & Anderson R.M. (2022). A Novel 2-Hit Zebrafish Model to Study Early Pathogenesis of Non-Alcoholic Fatty Liver Disease. Biomedicines, 10(2), 479.
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