Harmony software version 4

Manufactured by PerkinElmer

Harmony software version 4.1 is a high-performance imaging and analysis platform for cell-based assays. It provides advanced tools for image acquisition, processing, and analysis to support a wide range of cell-based applications.

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Lab products found in correlation

Operetta, by PerkinElmer (1 mentions) 96 well plate, by Corning (1 mentions) Operetta high content imaging system, by PerkinElmer (1 mentions) Cellcarrier, by PerkinElmer (1 mentions) Operetta high content screening system, by PerkinElmer (1 mentions) Sc 53140, by Santa Cruz Biotechnology (1 mentions) A11012, by Thermo Fisher Scientific (1 mentions) A11017, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Harmony software (213 citations) Harmony 4.9 (102 citations) Harmony software 4.8 (11 citations) Harmony High Content Imaging and Analysis Software Version 4.1 (2 citations)

3 protocols using harmony software version 4

High Content Imaging of Transfected Cells

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Microplates with seeded and transfected cells (Corning 96-well plate) were imaged on the High Content Screening System Operetta™ (PerkinElmer). In each well, images were acquired in 9 preselected fields with LWD 10x objective over four channels: brightfield, digital phase contrast (DPC) based on brightfield images, with excitation filters 460–490 and 520–550 nm and emission filters 500–550 and 560–630 nm for GFP and mCherry reporters, respectively. For the feature extraction, the images were analysed by Harmony software version 4.1 (PerkinElmer). Briefly, individual cell nuclei were segmented in the DPC channel. Nucleus morphology, GFP, and mCherry mean intensity were quantified in the cell nuclei population. Single-cell object features were extracted from each sample well. To discriminate between GFP/mCherry negative and positive cells, a threshold was determined based on the GFP/mCherry intensity frequency distribution of all samples for each experiment.

Ambrosini C., Destefanis E., Kheir E., Broso F., Alessandrini F., Longhi S., Battisti N., Pesce I., Dassi E., Petris G., Cereseto A, & Quattrone A. (2022). Translational enhancement by base editing of the Kozak sequence rescues haploinsufficiency. Nucleic Acids Research, 50(18), 10756-10771.

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Image Analysis Techniques for Neuroscience

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Microscopy images were visualized with Adobe Photoshop 2022, Fiji 2.9.0 or Imaris software version 9.9.1. Morphological reconstruction of neurons was performed using Imaris software version 9.9.1. Ca²⁺ imaging analysis was performed using MATLAB software. Quantification of immunofluorescence images was performed in FIJI (ImageJ) version 2.9.0 or using the Operetta high content imaging system coupled with Harmony software version 4.1 (PerkinElmer).

Ciceri G., Baggiolini A., Cho H.S., Kshirsagar M., Benito-Kwiecinski S., Walsh R.M., Aromolaran K.A., Gonzalez-Hernandez A.J., Munguba H., Koo S.Y., Xu N., Sevilla K.J., Goldstein P.A., Levitz J., Leslie C.S., Koche R.P, & Studer L. (2024). An epigenetic barrier sets the timing of human neuronal maturation. Nature, 626(8000), 881-890.

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Neurite Outgrowth Analysis of Differentiated NSC-34 Cells

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NSC-34 cells were fixed after seven days of differentiation and stained with Hoechst and mouse anti-Tubulin antibody (1:800; sc-53140, Santa Cruz). For HuD overexpressing cells, an additional immunostaining with a rabbit anti-HA antibody (1:600; A190-108A, Bethyl Laboratories) was performed. The following secondary antibodies, diluted 1:800, were then used: goat anti-rabbit Alexa Fluor 594 (A11012, Thermo Fisher Scientific), goat anti-mouse Alexa Fluor 488 (A11017, Thermo Fisher Scientific) and goat anti-mouse Alexa Fluor 594 (A11020, Thermo Fisher Scientific).
Neurite outgrowth was then analyzed on tubulin positive cells by High Content Screening System Operetta (PerkinElmer). Briefly, plates (96-well CellCarrier, PerkinElmer) were imaged and acquired in preselected fields with LWD 20x objective. For the feature extraction, the images were analyzed by Harmony software version 4.1 (PerkinElmer). Based on the Hoechst dye cell nuclei were identified. Starting from the cell body region, neurites were then detected in tubulin positive cells. The building block “Find Neurites” automatically calculated for each cell a set of neurite properties.

Tebaldi T., Zuccotti P., Peroni D., Köhn M., Gasperini L., Potrich V., Bonazza V., Dudnakova T., Rossi A., Sanguinetti G., Conti L., Macchi P., D’Agostino V., Viero G., Tollervey D., Hüttelmaier S, & Quattrone A. (2018). HuD Is a Neural Translation Enhancer Acting on mTORC1-Responsive Genes and Counteracted by the Y3 Small Non-coding RNA. Molecular Cell, 71(2), 256-270.e10.

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