Phenix high content screening system

Manufactured by Zeiss

The Phenix High Content Screening System is a laboratory instrument designed for automated, high-throughput image-based analysis of cells and other biological samples. It combines advanced microscopy, liquid handling, and data analysis capabilities to enable researchers to conduct comprehensive cell-based assays and screen large compound libraries efficiently.

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Lab products found in correlation

Lsm 800 microscope, by Zeiss (1 mentions) Glass microscope slide, by Thermo Fisher Scientific (1 mentions) Mouse anti tuj1, by Promega (1 mentions) Diamidino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions) Prolong diamond antifade mountant, by Thermo Fisher Scientific (1 mentions) Rat anti ctip2, by Abcam (1 mentions) Z0334, by Agilent Technologies (1 mentions) Goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)

2 protocols using phenix high content screening system

Immunofluorescent Staining of Neuronal Markers

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At 100 DIV, cells were fixed in 4 % (wt/vol) paraformaldehyde (PFA) for 10 min before permeabilization with 0.3 % (vol/vol) Triton-X-100 in PBS for 10 min and blocking with 5 % BSA in PBS for 30 min at RT. After blocking, cells were incubated with rat anti-Ctip2 (1:300; Abcam) and mouse anti-Tuj1 (1:1000; Promega) primary antibodies diluted in the blocking solution overnight at 4°C in a humid chamber. The following day, primary antibodies were removed, and cells were washed with PBS three times for 5 min. After the last wash, the cells were incubated with Alexa Fluor 488 anti-rat and Alexa Fluor 647 anti-mouse (1:1000) secondary antibodies diluted in a blocking buffer for 1h at RT protected from light. Secondary antibodies were then removed, and cells were washed twice with PBS and then counterstained with diamidino-2- phenylindole (DAPI) (Thermo Fisher) at 0.1 μg/ml for 10min at RT and washed three times with PBS. Cells were mounted on microscope glass slides (Thermo Fisher) with ProLong^™ Diamond Antifade Mountant (Thermo Fisher). For high content imaging, plates were stored in PBS. Stained cells were imaged using a Zeiss LSM 800 microscope and analysed using Fiji software or for quantification, cells were imaged on the Opera Phenix High Content Screening System (Figure 6E).

Pantazis C.B., Yang A., Lara E., McDonough J.A., Blauwendraat C., Peng L., Oguro H., Kanaujiya J., Zou J., Sebesta D., Pratt G., Cross E., Blockwick J., Buxton P., Kinner-Bibeau L., Medura C., Tompkins C., Hughes S., Santiana M., Faghri F., Nalls M.A., Vitale D., Ballard S., Qi Y.A., Ramos D.M., Anderson K.M., Stadler J., Narayan P., Papademetriou J., Reilly L., Nelson M.P., Aggarwal S., Rosen L.U., Kirwan P., Pisupati V., Coon S.L., Scholz S.W., Priebe T., Öttl M., Dong J., Meijer M., Janssen L.J., Lourenco V.S., van der Kant R., Crusius D., Paquet D., Raulin A.C., Bu G., Held A., Wainger B.J., Gabriele R.M., Casey J.M., Wray S., Abu-Bonsrah D., Parish C.L., Beccari M.S., Cleveland D.W., Li E., Rose I.V., Kampmann M., Aristoy C.C., Verstreken P., Heinrich L., Chen M.Y., Schüle B., Dou D., Holzbaur E.L., Zanellati M.C., Basundra R., Deshmukh M., Cohen S., Khanna R., Raman M., Nevin Z.S., Matia M., Van Lent J., Timmerman V., Conklin B.R., Chase K.J., Zhang K., Funes S., Bosco D.A., Erlebach L., Welzer M., Kronenberg-Versteeg D., Lyu G., Arenas E., Coccia E., Sarrafha L., Ahfeldt T., Marioni J.C., Skarnes W.C., Cookson M.R., Ward M.E, & Merkle F.T. (2022). A reference human induced pluripotent stem cell line for large-scale collaborative studies. Cell stem cell, 29(12), 1685-1702.e22.

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Immunostaining and Imaging of Neuronal Cells

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Cells were fixed for 10 min in 4% paraformaldehyde and 4% sucrose in PBS and then for 10 min in pre-chilled methanol. Cells were simultaneously permeabilized and blocked using PBS containing 2% normal goat serum and 0.1% Triton-X-100 for 1 h at room temperature. Immunolabelling was performed according to previously published protocols [54 ]. Primary antibodies were against tau (Rabbit IgG, #A0024, DAKO, 1/500), MAP2 (Mouse IgG, #GTX8266, GeneTex, 1/200), and GFAP (Rabbit IgG, #Z0334, DAKO, 1/100), and the appropriate species of AlexaFluor-conjugated secondary antibodies (Goat anti-rabbit IgG or Goat anti-mouse IgG, Life Technologies). High-resolution digital images were acquired using the Opera Phenix High Content Screening System in confocal mode with a Zeiss 20 × water NA 1.0 objective. The fluorophores were detected with the following excitation and emission (Ex/Em) wavelengths: Hoechst 33342 (405/435–480), AF488 (488/500–550), and far red (wavelengths above 655 nm). Cell segmentation and quantification analyses were performed using the Harmony software, version 4.9. The experimenter was blinded to treatment.

Perez-Nievas B.G., Johnson L., Beltran-Lobo P., Hughes M.M., Gammallieri L., Tarsitano F., Myszczynska M.A., Vazquez-Villasenor I., Jimenez-Sanchez M., Troakes C., Wharton S.B., Ferraiuolo L, & Noble W. (2021). Astrocytic C–X–C motif chemokine ligand-1 mediates β-amyloid-induced synaptotoxicity. Journal of Neuroinflammation, 18, 306.

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