HepAD38 cells (1 χ 104) were seeded in 96-well 2% collagen-coated image plates (BD Falcon) in tet media. The next day, the cells were washed twice with PBS, and complete media containing compounds was added to the wells with 1% final DMSO concentration. At 4 days post-induction, cells were fixed at room temperature (RT) for 15 min in 4% formaldehyde, permeabilized with 0.1% Triton X-100 in PBS, and blocked with 10% goat serum (Jackson ImmunoResearch) and 1% bovine serum albumin (BSA) in PBS. Rabbit anti-HBV core primary antibody (Dako) (1:1,000) was bound overnight at 4°C. Samples were incubated at RT for 45 min with 1:2,000 goat antirabbit Alexa Fluor 568 secondary antibody (Invitrogen) and stained for nuclei using Hoechst 33342 (Invitrogen) or DRAQ5 (Thermo Scientific). Images were taken on a confocal laser scanning Leica TCS SP8 microscope.
Collagen coated image plates
Manufactured by BD
2% collagen-coated image plates are a type of laboratory equipment used for cell culture and imaging applications. They provide a surface coated with a 2% collagen solution, which can facilitate cell attachment and growth. The core function of these plates is to serve as a substrate for culturing cells and conducting various microscopic observations and analyses.
Automatically generated - may contain errors
Lab products found in correlation
Tcs sp8 microscope, by Leica (2 mentions)
Lsm 510 meta confocal microscope, by Zeiss (2 mentions)
Goat anti rabbit alexa fluor 568 secondary antibody, by Thermo Fisher Scientific (1 mentions)
Goat serum, by Jackson ImmunoResearch (1 mentions)
Rabbit anti hbv core primary antibody, by Agilent Technologies (1 mentions)
Draq5, by Thermo Fisher Scientific (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
2 protocols using collagen coated image plates
1
HBV Core Protein Immunostaining Assay
2
HepAD38 Cells Imaging and Analysis
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Example 10
HepAD38 cells (1×104) were seeded in 96-well 2% collagen-coated image plates (BD Falcon, Bedford, Mass.) in tet media. On the appropriate days, the cells were washed twice with PBS and complete media containing compounds was added to the wells with 1% final DMSO concentration. Cells were fixed and stained on the indicated days post-induction, and the protein redistribution was analyzed from confocal images taken with a Zeiss LSM 510 Meta confocal microscope with Autostage, Multitile, and MultiTime series 4.0.31 beta software, as previously described (Liu, et al., Antimicrob. Agents Chemother. 59:3482-3492, 2015). Imaging was carried out using a 40× objective, capturing 9 images per well. Images were processed using CellProfiler software (Kamentsky, et al., Bioinformatics 27:1179-1180, 2011; Carpenter, et al., Genome Biology 7:R100, 2006; Lamprecht, et al., BioTechniques 42:71-75, 2007). Additional images were taken on a confocal laser scanning Leica TCS SP8 microscope.
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