Lsm 710 confocal microscope

Manufactured by PerkinElmer

Sourced in United Kingdom

The LSM 710 is a confocal microscope designed for high-resolution imaging and analysis of biological samples. It features a laser-scanning system that allows for the acquisition of optical sections through a specimen, enabling 3D reconstruction and visualization of complex structures.

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Lab products found in correlation

Volocity software, by PerkinElmer (2 mentions) Volocity 6, by PerkinElmer (2 mentions) Axiocam cm1, by Zeiss (2 mentions) Axio lab a1, by Zeiss (2 mentions) Diaphot inverted microscope, by Nikon (1 mentions) Volocity 3d image analysis software, by PerkinElmer (1 mentions) Bubr1, by BD (1 mentions) Mps1 pt33ps37, by Thermo Fisher Scientific (1 mentions) Zwint 1, by Abcam (1 mentions) Mitotracker green fm, by Thermo Fisher Scientific (1 mentions) Mouse anti ha, by Merck Group (1 mentions) Goat serum, by Thermo Fisher Scientific (1 mentions) Doxycycline, by Merck Group (1 mentions) Lab tek 2, by Thermo Fisher Scientific (1 mentions) Yh2ax, by Merck Group (1 mentions)

5 protocols using lsm 710 confocal microscope

Immunofluorescence Assay for Mitotic Regulators

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For immunofluorescence, cells were pretreated with CCT271850 for 1 h, then treated with nocodazole, MG132 and CCT271850 for additional 1 h. Cells were fixed and stained according to the protocol previously described (Gurden et al, 2015 (link)). Primary antibodies used were as follows: anticentromere antibodies (ACA; ImmunoVision, Springdale, AR, USA, HCT-0100), BubR1 (BD Biosciences, San Jose, CA, USA, 612503), Mad1 (Abcam, Cambridge, UK, ab45286), Mad2 (Bethyl Laboratories Inc., Montgomery, TX, USA, A300-301A), MPS1 (Invitrogen, 35–9100), MPS1 pT33pS37 (Life Technologies, 44–1325 G) and Zwint-1 (Abcam, ab84367). Images were acquired using a Zeiss LSM 710 confocal microscope and processed using the Volocity 3D Image analysis software (PerkinElmer). Time-lapse microscopy was performed in 96-well Ibidi plate (Thistle Scientific, Glasgow, UK) using a Diaphot inverted microscope (Nikon, Tokyo, Japan), in a humidified CO₂ chamber at 37 °C, using a motorised stage (Prior Scientific, Cambridge, UK), controlled by Simple PCI software (Compix, Irvine, CA, USA).

Faisal A., Mak G.W., Gurden M.D., Xavier C.P., Anderhub S.J., Innocenti P., Westwood I.M., Naud S., Hayes A., Box G., Valenti M.R., De Haven Brandon A.K., O'Fee L., Schmitt J., Woodward H.L., Burke R., vanMontfort R.L., Blagg J., Raynaud F.I., Eccles S.A., Hoelder S, & Linardopoulos S. (2017). Characterisation of CCT271850, a selective, oral and potent MPS1 inhibitor, used to directly measure in vivo MPS1 inhibition vs therapeutic efficacy. British Journal of Cancer, 116(9), 1166-1176.

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Mitochondrial Labeling and Immunostaining

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Procyclic or metacyclic promastigote cells were incubated in culture with 200 nM of MitoTracker™ Green FM (Molecular Probe) for 30 min at 26°C, harvest and antibody stained as described (30 (link)) using mouse anti-HA [1:500] (Sigma) or rabbit anti-TbRBP16 [1:500] (29 (link)) and Alexa secondary antibodies [1:10 000]. Images were acquired on a Zeiss LSM 710 confocal microscope and images were processed using PerkinElmer Volocity software.

Ferreira T.R., Dowle A.A., Parry E., Alves-Ferreira E.V., Hogg K., Kolokousi F., Larson T.R., Plevin M.J., Cruz A.K, & Walrad P.B. (2020). PRMT7 regulates RNA-binding capacity and protein stability in Leishmania parasites. Nucleic Acids Research, 48(10), 5511-5526.

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Radiation-Induced DNA Damage Analysis

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T98-EGFP and T98-EGFP-8.6 cells were grown on Lab-Tek II (Thermo Scientific) glass chamber slides (using the aforementioned conditions). Doxycycline (1 μg mL⁻¹, Sigma) was added 24 h after seeding. Seven days after the addition of Doxycycline, cells were treated with 5 Gy of radiation using an MDS Nordion Gammacell 40 research irradiator. After treatment, cells were fixed in 4% (v/v) paraformaldeyde, permeabilized for 10 min with 0.5% (v/v) Triton-X 100 in phosphate-buffered saline, and blocked with a 1× casein (Sigma) or 3% (v/v) goat serum (Gibco) mixture for 2 h at room temperature (RT). Cells were then exposed to a 1:500 primary antibody solution (either y-H2AX (Millipore) or Rad-51 (Calbiochem)) overnight at 4 °C. The slides were then washed with phosphate-buffered saline and exposed to a 1:500 Alexa-Fluor 594 secondary antibody solution. Slides were washed again with phosphate-buffered saline, and nuclei were counterstained with VectaShield mounting media with DAPI. Images were acquired using a Zeiss LSM 710 confocal microscope and analyzed (including foci counting) with Perkin Elmer’s Volocity software.

White E.R., Sun L., Ma Z., Beckta J.M., Danzig B.A., Hacker D.E., Huie M., Williams D.C., Edwards R.A., Valerie K., Mark Glover J.N, & Hartman M.C. (2015). Peptide Library Approach to Uncover Phosphomimetic Inhibitors of the BRCA1 C-Terminal Domain. ACS chemical biology, 10(5), 1198-1208.

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Neurite Growth Quantification in 2D and 3D Co-Cultures

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Fluorescence microscopy (Zeiss Axiolab A1, Axiocam Cm1) was used to capture images of neurites from five pre‐determined fields of each gel or coverslip using a ×20 lens. The neurite number in the determined fields depended on the cell type and model used (~1–12 neurites). Briefly, the positions of the pre‐determined fields on coverslips were equally spaced (625‐μm apart) in a line across the diameter at the center of the coverslip, for the gels the same method was used but the line of fields was along the edge of the construct where alignment was greatest. The length of each neurite in each field was measured using ImageJ. Following stabilization the gel acquired a thickness of 100 μm and the neurons extended predominantly in a single horizontal plane along the top surface, following the aligned Schwann cells (Phillips, 2014). This allowed analysis of neurite growth to be comparable between both the monolayer and 3D co‐cultures.
Tile scans were used to capture high‐magnification (x20) micrographs from the entire nerve cross‐section using a Zeiss LSM 710 Confocal microscope and images were analyzed using Volocity™ 6.4 (PerkinElmer) running automated image analysis protocols to determine the number of neurofilament‐immunoreactive neurites in each transverse nerve section.

RAYNER M.L., LARANJEIRA S., EVANS R.E., SHIPLEY R.J., HEALY J, & PHILLIPS J.B. (2018). Developing an In Vitro Model to Screen Drugs for Nerve Regeneration. Anatomical Record (Hoboken, N.j. : 2007), 301(10), 1628-1637.

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Quantifying Neurite Outgrowth via Fluorescence Microscopy

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Fluorescence microscopy (Zeiss Axiolab A1, Axiocam Cm1, Carl Zeiss GmbH) was used to capture images of neurites from five pre-determined fields of each gel using a ×20 lens. The positions of the pre-determined fields on the gels were equally spaced (625 μm apart) in a line along the edge of the construct where alignment was greatest [27 (link)]. The length of each neurite in each field (~1–12 neurites) was measured using Fiji ImageJ [29 (link)]. Following stabilization, the gel acquired a thickness of 100 μm and the neurons extended predominantly in a single horizontal plane along the top surface, following the aligned Schwann cells [30 (link)].
Tile scans were used to capture high-magnification (×20) micrographs from the entire nerve cross-section using a Zeiss LSM 710 confocal microscope and images were analysed using Volocity™ 6.4 (PerkinElmer, Beaconsfield, UK) running automated image analysis protocols to determine the number of neurofilament-immunoreactive neurites in each transverse nerve section.

Rayner M.L., Kellaway S.C., Kingston I., Guillemot-Legris O., Gregory H., Healy J, & Phillips J.B. (2022). Exploring the Nerve Regenerative Capacity of Compounds with Differing Affinity for PPARγ In Vitro and In Vivo. Cells, 12(1), 42.

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