Ix 81 high content screening inverted microscope

Manufactured by Olympus

Sourced in Japan

The IX-81 is a high content screening inverted microscope designed for cell-based assays. It features an automated stage, autofocus, and imaging capabilities to capture high-quality images of cell samples. The IX-81 is equipped with a variety of fluorescence detection channels and illumination sources to support a range of experimental applications.

Automatically generated - may contain errors

Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) 48 well culture plate, by Thermo Fisher Scientific (1 mentions) Phalloidin alexa488, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

IX inverted microscope 1×81 inverted microscope Inverted microscope

4 protocols using ix 81 high content screening inverted microscope

Keratinocyte Adhesion and Migration Assay

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Keratinocytes were harvested and seeded (1 × 10⁴ cells/well) on the various substrates in the wells of a 48-well culture plate (NUNC). After three days of culture in DKSFM keratinocytes were fixed with 4% paraformaldehyde/PBS for 15 min at RT, incubated in PBS/1%BSA for 1 h at RT before polymerized actin was stained with 1 unit/ml of phalloidin-Alexa 488 (Molecular Probe) and nuclei with DAPI. Using a 20x objective and an Olympus IX-81 high content screening inverted microscope (Olympus), 8 by 8 non-overlapping quadrants were imaged. Cell size as delineated by the stained actin cytoskeleton was determined using the Cell Profiler software^{75 (link)}.
Keratinocyte movement was assessed on either Fib-Mat, collagen I coating or uncoated TCP. Keratinocytes were seeded as described and following an overnight incubation to allow cell adhesion; live cell images were collected using Olympus IX-81 high content screening inverted microscope (Olympus). Time-lapse images were taken at 15-minute intervals over 2 days using a 10x objective.

Wong C.W., LeGrand C.F., Kinnear B.F., Sobota R.M., Ramalingam R., Dye D.E., Raghunath M., Lane E.B, & Coombe D.R. (2019). In Vitro Expansion of Keratinocytes on Human Dermal Fibroblast-Derived Matrix Retains Their Stem-Like Characteristics. Scientific Reports, 9, 18561.

+ Open protocol

+ Expand

Keratinocyte Proliferation Assay Using Ki67

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Briefly, keratinocytes were harvested and then seeded as described above and at day 3, keratinocytes were fixed with 4% paraformaldehyde for 15 min at RT, permeabilized with cold 0.1% Triton X-100/PBS for 3 min and then incubated in PBS/1% BSA for 1 h at RT. The keratinocytes were blocked with 10% goat serum/1% BSA/PBS for 1 h at RT before being incubated with a 0.3 µg/ml anti-Ki67 antibody (Novacastra). This was followed by a 1 h incubation with Alexa 488 conjugated anti-mouse antibody at RT. Keratinocyte nuclei were stained using DAPI. Images were taken on the Olympus IX-81 high content screening inverted microscope. Using a 10x objective, 7 by 11 non-overlapping quadrants were imaged. The percent of keratinocytes positive for Ki67 was determined using the Cell Profiler software^{75 (link)}.

+ Open protocol

+ Expand

Automated Keratin Aggregate Quantification

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fluorescent images of EBS cells with keratin aggregates were acquired using the Olympus IX-81 high content screening inverted microscope (Olympus, Tokyo, Japan), attached with a Marzhauser Scan IM (120 × 100) motorized XY stage controlled by Ludl Mac5000 and a Photometrics CCD camera (CoolSNAP HQ2) with DAPI, EGFP, and TRITC filters. Fluorescent images were analyzed with customized algorithms written with ImageJ software for quantitative analysis of keratin aggregates in mutant cells (Tan et al., 2021 (link)). Alternatively, these images were analyzed using CellProfiler free open-source software (cellprofiler.org) to count keratin aggregates in mutant cells. Briefly, a custom pipeline was designed and applied to the images to identify three key features of a cell: the nucleus, plasma membrane, and keratin aggregates. These features are needed to determine the overall cell density (nuclear count), cell boundary (plasma membrane), and the number of keratin aggregates (GFP-tagged K14 protein) present in each cell identified within the cytoplasm. The CellProfiler software calculates the average number of keratin aggregates per cell and the percentage of cells containing keratin aggregates for each set of fluorescent images.

Badowski C., Tan T.S., Aliev T., Trudil D., Larina M., Argentova V., Firdaus M.J., Benny P., Woo V.S, & Lane E.B. (2022). Detrimental Effects of IFN-γ on an Epidermolysis Bullosa Simplex Cell Model and Protection by a Humanized Anti–IFN-γ Monoclonal Antibody. JID Innovations, 2(2), 100096.

+ Open protocol

+ Expand

Keratinocyte Proliferation on Substrates

Check if the same lab product or an alternative is used in the 5 most similar protocols

The proliferation of keratinocytes on the substrates (Fib-Mat, collagen I (3 µg/cm2) and tissue culture plastic (TCP)) was assessed. Keratinocytes were harvested and seeded at a density of 1 × 10⁴ cells/well in a 48-well tissue culture plate (NUNC, ThermoFisher Scientific) and grown for six days. At 24 h intervals keratinocytes were fixed for 5 min with cold acetone:methanol (1:1), washed with PBS and incubated with PBS/1%BSA for 1 h at RT before the nuclei were stained with DAPI (Sigma). Using an Olympus IX-81 high content screening inverted microscope (Olympus; Tokyo, Japan) and a 10x objective, 7 by 11 non-overlapping quadrants were imaged, to produce a 0.5 cm² area image. Nuclei/cell numbers were determined using Fiji-Image J software^{74 (link)} and its “Find Object” macro.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!