Cell observer imaging

Manufactured by Zeiss

The Zeiss Cell Observer is a versatile imaging system designed for high-resolution live-cell microscopy. It provides reliable and precise visualization of cellular processes and dynamics, enabling researchers to capture and analyze complex biological phenomena. The system's core function is to facilitate the acquisition of high-quality, time-lapse images and videos of living cells, supporting a wide range of applications in cell biology, developmental biology, and biomedical research.

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Lab products found in correlation

Protease inhibitor cocktail, by Roche (1 mentions) Falcon cell culture insert, by BD (1 mentions) 710 confocal microscopy, by Zeiss (1 mentions)

Close spelling variants of this product

Cell Observer (142 citations) Zeiss Cell Observer Live Imaging microscope (8 citations) Axio Observer Z1 Live-Cell imaging system (5 citations)

3 protocols using cell observer imaging

Cell Migration and Invasion Assays

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Cell migration and invasion assays were carried out in a 24-well trans-well plate with an 8-µm Falcon cell culture insert (Becton-Dickinson) to separate the lower and upper culture chambers. In brief, C4-2 cells or infected LNCaP cells with empty-vector, SKP2-WT, or SKP2-KQKQ lentivirus were plated in the upper chamber at 10⁵ cells per well in hormone-free RPMI (without phenol red), with the bottom chamber containing DHT (100 nM). Cells were allowed to migrate for 48 hours and invade for 72 hours. Cells that migrated or invaded to the lower surface of the filter were fixed with 20% (w/v) methanol and stained with 0.5% (w/v) crystal violet. Cells were counted in three random fields under a microscope (Zeiss Cell Observer imaging). Therefore, the invasion assay was similar to the cell migration assay, except that the membrane filter was precoated with diluted Matrigel with serum and phenol-free RPMI (1:3). For the wound-healing assay, C4-2 cells treated with inhibitors along with DMSO (vehicle) in hormone-free RPMI were seeded overnight in 12-well plates in triplicate, and confluent cells were subjected to an in vitro scratch for wound healing analysis using Zeiss microscopy.

Rezaeian A.H., Phan L.M., Zhou X., Wei W, & Inuzuka H. (2023). Pharmacological inhibition of the SKP2/p300 signaling axis restricts castration-resistant prostate cancer. Neoplasia (New York, N.Y.), 38, 100890.

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Protein Interaction and Localization Assays

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IP, IB, IF and IHC assays were done as described previously, with brief modifications⁴⁶. For protein-protein interactions (IP), cells were subjected to lysis by E1A lysis buffer (250 mM NaCl, 50 mM HEPES [pH 7.5], 0.1% NP- 40, 5 mM EDTA, protease inhibitor cocktail [Roche]). For IF, cells were grown on chamber slides. After incubation in normoxic or hypoxic (1% oxygen) conditions for 12 h, cells were washed and fixed with 4% paraformaldehyde and subjected to permeabilization in 0.3% Triton X-100 and 5% glycine in phosphate-buffered saline solution (PBS) on ice. Cells were blocked in 4% bovine serum albumin (BSA) and immunolabelled with specific antibodies, then observed by Zeiss Cell Observer imaging and Zeiss 710 confocal microscopy. The list of main commerial antibodies with identifications, applications and dilutions were included in Supplementary Table 6. The anti-SHARP1 antibody was a gift from Dr. S. Piccolo (IB: 1:1000). Each image resulted from western blots and immunofluorescence represents >=3 independent experiments.

Rezaeian A.H., Li C.F., Wu C.Y., Zhang X., Delacerda J., You M.J., Han F., Cai Z., Jeong Y.S., Jin G., Phan L., Chou P.C., Lee M.H., Hung M.C., Sarbassov D, & Lin H.K. (2016). A hypoxia-responsive TRAF6-ATM-H2A.X signalling axis promotes HIF1α activation, tumorigenesis and metastasis. Nature cell biology, 19(1), 38-51.

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Cell Migration and Invasion Assays

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The cell migration and invasion assays were performed in a 24-well transwell plate with 8-μm polyethylene terephthalate membrane filters (Falcon cell culture insert; Becton-Dickinson) separating the lower and upper culture chambers. In brief, MDA-MB-231 or A549 cells in which H2AX had been knocked down and restored were plated in the upper chamber at 10⁵ cells per well in serum-free DMEM, while the bottom chamber contained DMEM with 10% FBS. Cells were allowed to migrate for 6 h. Cells that migrated to the lower surface of the filter were fixed with 4% (w/v) paraformaldehyde and stained with 0.5% (w/v) crystal violet. Cells were counted in three random fields by Zeiss Cell Observer imaging. Invasion assay of MDA-MB-231 cells was essentially similar to the cell migration assay, except that the membrane filter was precoated with diluted Matrigel (Matrigel:serum-free DMEM, 1:3) before the assay and the incubation time was 16 h. For the wound healing assay, MDA-MB- 231 cells in which H2AX was knocked down and then rescued with wild-type or mutant H2AX were seeded overnight in 12-well plates in triplicate. Confluent cells were subjected to an in vitro scratch, and wound healing was recorded by Zeiss 710 confocal microscopy.

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