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Live cell observer

Manufactured by Zeiss
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The Live Cell Observer is a microscope system designed for high-resolution imaging of living cells. It provides a controlled environment for maintaining cell cultures during observation.

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Lab products found in correlation

Axio zoom v16, by Zeiss (2 mentions) Eight well chamber slide, by Ibidi (1 mentions) Las x software, by Leica (1 mentions) Tc sp8 confocal microscope, by Leica (1 mentions) Lsm 710 inverted confocal microscope, by Zeiss (1 mentions) Anti cd117 beads, by Miltenyi Biotec (1 mentions) Axiovert 200m inverted microscope, by Zeiss (1 mentions) Celltracker green dye, by Thermo Fisher Scientific (1 mentions) Axiocam mrm camera, by Zeiss (1 mentions) A1r inverted confocal microscope, by Nikon (1 mentions)

Close spelling variants of this product

Axio Observer Z1 Live-Cell imaging system Live Cell Observer/Deconvolution system Live Cell Observer microscope Axio Observer Live Cell microscope Zeiss Cell Observer Live Imaging microscope Cell Observer

5 protocols using live cell observer

1

Detailed Invasion Assay Protocol

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A detailed protocol for assaying invasion is available in S1 Text. The invasion assay proceeded largely as previously described [59 (link)]. Briefly, parasite lines were treated ±ATc and resuspended in IC buffer, to prevent invasion, and spun down onto the HFF monolayer in an eight-well chamber slide (Ibidi). IC buffer was exchanged for DME/1% FCS/10 mM HEPES, pH 7.5, and incubated for 10 minutes at 37 °C. Samples were then fixed, stained with anti-SAG1 mAb DG52 (1:3,000 dilution), permeabilised, and stained with rabbit anti-GAP45 (1:2,000) [58 (link)], followed by secondary antibodies, then imaging on a Zeiss Live Cell Observer and quantitation by manual counting. For invasion assays using GFP-expressing parasites, GAP45 staining was not carried out. Invasion assays with DCCD were carried out as described above except that parasites were allowed to invade in D1/HEPES medium containing 50 μM DCCD (Sigma). Graphs were generated and statistical tests performed using Prism 7 software.
Uboldi A.D., Wilde M.L., McRae E.A., Stewart R.J., Dagley L.F., Yang L., Katris N.J., Hapuarachchi S.V., Coffey M.J., Lehane A.M., Botte C.Y., Waller R.F., Webb A.I., McConville M.J, & Tonkin C.J. (2018). Protein kinase A negatively regulates Ca2+ signalling in Toxoplasma gondii. PLoS Biology, 16(9), e2005642.
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2

Imaging Primary Chicken Lung Slices

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The PCLS isolated from CSF1R-eGFP or CSF1R-mApple transgenic birds were imaged floating in DMEM within a 35 mm µ-dish (Ibidi), weighed down with sterile 13 mm coverslips (Scientific Laboratory Supplies Ltd, Coatbridge, UK). Live images were captured using a Zeiss Live Cell Observer with the heated stage set to 37 °C and the imaging chamber set to 37 °C and 5% CO2. PCLS at 3 days post slice were incubated with either red fluorescent 1 μm latex beads (Thermo Fisher Scientific, Paisley, UK) or 5 × 109 CFU/mL APEC O1-eGFP.
Images of PCLS were also captured using a Zeiss Axio Zoom.v16 in the wells of a 24 well plate at 7X or 16X magnification. Confocal images were captured using a Leica TCS P8 confocal microscope at 200X or 630X magnification and the LAS X software (Leica, Wetzler, Germany) or a Zeiss LSM710 inverted confocal microscope and Zen software (Zeiss).
Bryson K.J., Garrido D., Esposito M., McLachlan G., Digard P., Schouler C., Guabiraba R., Trapp S, & Vervelde L. (2020). Precision cut lung slices: a novel versatile tool to examine host–pathogen interaction in the chicken lung. Veterinary Research, 51, 2.
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3

Imaging Hematopoietic Stem Cell Differentiation

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Picovitro slides (Picovitro) were first coated with Collagen type I, followed by culture of OP9 stromal cells, previously irradiated with 30 Gy, at a number of 102 per well in dendritic cell conditioned medium [4 (link)]. Cultures were left overnight for OP9 adherence. The following day, HSCs were prepared from c-kit (CD117)-enriched BM from either C57BL/6, UBC-GFP mice using anti-CD117 beads (Miltenyi), followed by staining with antibodies against Sca-1, ckit and IL-7R. Progenitors were sorted as Sca-1+c-kithiIL7R-progenitors [5 (link)]. PDMS membranes were layered on top, and slides fixed to a custom-bracket to fully encapsulate slides in order to prevent dehydration but allow gas exchange. The slide and bracket were mounted in a Zeiss Live Cell Observer, with 5% CO2-in-air, and at 37° C. Phase contrast images of multiple positions in the slide were taken with a 10x Plan-Neofluar phase contrast objective with a 1.6x Optovar every 2 minutes. GFP images were captured every hour, with each clone being GFP+ or GFP- throughout the image sequence. The GFP information was used for a separate validation experiment, although it was helpful during validation here for resolving tracking ambiguities.
Mankowski W.C., Winter M.R., Wait E., Lodder M., Schumacher T., Naik S.H, & Cohen A.R. (2014). Segmentation of Occluded Hematopoietic Stem Cells from Tracking. Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual Conference, 2014, 5510-5513.
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4

Monitoring Cell Death in U937 Cells

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U937 cells were plated in a black 96 well optical bottom plate (5×104 cells/well) together with DMSO, peptides (5 μM) +/− Q-VD-OPh (50 μM) in phenol-red and serum-free RPMI medium (final volume 100 μL) containing CellTracker Green dye (Life Technologies, C2925) and propidium iodide (2 μg/ml). Cells were imaged every 30 minutes for 24 hours on a Zeiss Live Cell Observer, consisting of an Axiovert 200M inverted microscope, equipped with environmental control and CO2, LED Colibri illumination, a 20×/0.8 NA objective and AxioCam MRm camera. Zeiss filter set Ex 470/40 and Em 525/50 together with LED module 470 was used for CellTracker Green fluorescence and Ex 560/40 and 630/75 together with LED module 540-580 for propidium iodide fluorescence.
Checco J.W., Lee E.F., Evangelista M., Sleebs N.J., Rogers K., Pettikiriarachchi A., Kershaw N.J., Eddinger G.A., Belair D.G., Wilson J.L., Eller C.H., Raines R.T., Murphy W.L., Smith B.J., Gellman S.H, & Fairlie W.D. (2015). α/β-Peptide Foldamers Targeting Intracellular Protein-Protein Interactions with Activity in Living Cells. Journal of the American Chemical Society, 137(35), 11365-11375.
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5

Imaging E13.5 TCF/Lef::H2B-GFP Skin Explants

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E13.5 TCF/Lef::H2B-GFP dorsal skin explants were dissected and imaged as described previously using a custom imaging chamber [81 (link)] with the following modifications. Instead of using a central imaging clip, the entire base of the chamber was filled with 1% (weight/volume) agarose in PBS. Dissected embryonic skin was mounted dermis-side down onto a black nitrocellulose filter membrane (Millipore) with a 45 μm pore size. The membrane was subsequently placed onto the agarose block, and a lummox membrane (Greiner) was clamped across it with an o-ring such that the skin was sandwiched between the 2 membranes. The imaging chamber was filled with DMEM without phenol red containing 4,500 mg/L glucose, 2% FBS, 1% penicillin/streptomycin, and 0.584 g/l L-glutamine. Images were captured with a 20X objective using a Nikon A1R inverted confocal microscope in a heated chamber supplied with 5% CO2 in air. Bead and FGF9/LDN193189 combination experiments were performed using a Zeiss Live Cell Observer and are described in S1 Supporting Methods.
Glover J.D., Wells K.L., Matthäus F., Painter K.J., Ho W., Riddell J., Johansson J.A., Ford M.J., Jahoda C.A., Klika V., Mort R.L, & Headon D.J. (2017). Hierarchical patterning modes orchestrate hair follicle morphogenesis. PLoS Biology, 15(7), e2002117.
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