An EdU kit (Cell‐Light EdU Apollo 643 In Vitro Imaging Kit; RiboBio) was used to evaluate the cell proliferation viability according to the manufacturer’s instructions. Images were detected and analyzed with a microscope at 200× (Olympus). The ratio of EdU‐stained cells (with red fluorescence) to Hoechst‐stained cells (with blue fluorescence) was used to evaluate cell proliferation activity.
Cell light edu apollo 643 in vitro imaging kit
Manufactured by RiboBio
Sourced in China
The Cell-Light EdU Apollo 643 In Vitro Imaging Kit is a laboratory equipment designed for detecting and visualizing DNA synthesis in proliferating cells. It utilizes a modified thymidine analog, EdU (5-ethynyl-2'-deoxyuridine), which is incorporated into newly synthesized DNA during cell division. The kit includes the necessary reagents and components to perform this analysis in an in vitro setting.
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Lab products found in correlation
Cck 8 solution, by Dojindo Laboratories (2 mentions)
Fluorescence photomicroscope, by Zeiss (2 mentions)
Fluorescent microscope, by Olympus (2 mentions)
Microscope at 200, by Olympus (1 mentions)
Fluoview fv1000 confocal laser, by Olympus (1 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions)
Axiophot photomicroscope, by Zeiss (1 mentions)
Edu cell proliferation assay kit, by RiboBio (1 mentions)
Operetta cls high content analysis system, by PerkinElmer (1 mentions)
96 well microplate, by Greiner (1 mentions)
Harmony high content imaging and analysis software, by PerkinElmer (1 mentions)
Cck 8 solution, by Beyotime (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Photomicroscope, by Zeiss (1 mentions)
16 protocols using cell light edu apollo 643 in vitro imaging kit
1
Cell Proliferation Assay with EdU
2
Evaluating HaCaT Cell Proliferation via EdU Assay
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To determine whether PPI affects HaCaT cell proliferation, an EdU incorporation assay was carried out. 48 hours after cell seeding, 10 μM EdU was poured into the culture following incubation for some time followed by fixation using 4% paraformaldehyde in PBS for 15 minutes. EdU labeling with an azide derivative of Apollo 643 was performed using a Cell-Light™ EdU Apollo 643 In Vitro Imaging Kit (#C10310-2, RiboBio Co., Ltd., China). A 652 nm laser was used for the excitation of Apollo 643. Microscopic images were obtained with a FluoView FV1000 confocal laser scanning microscope (Olympus, Japan). ImageJ (National Institutes of Health, Bethesda, MD, USA) was used to get composite images.
3
Quantifying Cell Proliferation Assays
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Cell proliferation was assessed using Cell Counting Kit-8 (CCK-8) and ethynyl deoxyuridine (EdU) incorporation assays. For CCK-8 assay, indicated PCa cells were plated at 2 × 103 cells/well into 96-well plates. After incubation for the indicated time, CCK-8 solution (Dojindo Laboratories, Kumamoto, Japan) was added into the wells. Optical density (OD) values at 450 nm at indicated time points were detected using an automatic enzyme-linked immune detector. EdU incorporation assay was carried out using the Cell-Light EdU Apollo 643 In Vitro Imaging Kit (RiboBio, Guangzhou, China) following the protocol. Results were detected with a Zeiss fluorescence photomicroscope (Carl Zeiss, Oberkochen, Germany) and counted based on five randomly chosen visual fields.
4
Cell Proliferation Assay Using EdU
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Logarithmically proliferating Lv-miR-340-A2780 or Lv-miR-340-SKOV3 cells were seeded in 96-well plates (8 × 104 cells/well) 12 h before staining with the Cell-Light™ EdU Apollo®643 In Vitro Imaging Kit (RiboBio) according to the manufacturer’s protocol. Briefly, the cells were incubated with 50 μM EdU for 2 h before fixation with 4% paraformaldehyde, permeabilization with 0.5% Triton X-100, and EdU staining. The cell nuclei were stained with Hoechst 33342 for 30 min. The number of EdU-positive cells in five random fields was counted under laser scanning confocal microscopy.
5
Evaluating Cell Proliferation Assay
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The cells were seeded into 96-well plates (3,000 cells per well) and performed according to protocols mentioned above. Edu Cell Proliferation Assay Kit (RiboBio, Cell-Light™EdU Apollo®643 In Vitro imaging kit) was used to assessed cell proliferation according to the manufacture’s instruction. The cells were added with 100 μL Edu (diluent reagent A with a complete medium by 1:1,000) and incubated for 2 h at 37°C. The cells were fixed in 4% paraformaldehyde for 15–30 min and incubated with 1× Apollo® solution for 30 min at room temperature. The cell nuclei were stained with 100 μL 1× Hoechst33342 for 30 min. Finally, the cells were examined under a fluorescent microscope (Olympus, Japan). Data were presented as a fold-change of Edu-incorporating cells compared with negative controls.
6
Cell Proliferation and Colony Formation
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For the colony formation assay, treated cells were seeded in six-well plates at a density of 1000 cells per well and cultured for 8–10 days. The colonies were then fixed with cold methanol and stained with 0.1% crystal violet; colonies comprising more than 50 cells were counted. For the EdU incorporation assay, cell proliferation was determined as described previously39 (link), using the Cell-Light™ EdU Apollo®643 In Vitro Imaging kit (RiboBio).
7
HeLa Cell Growth and DNA Synthesis
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For growth curves, HeLa cells were seeded in 96-well plates treated with or without 10 μM OA and were imaged in an IncuCyte (Essen, USA) automated incubator microscope. Pictures were taken every 4 h., and cell confluence was calculated per well using an associated software algorithm. DNA synthesis was determined using a Cell-Light EdU Apollo 643 In Vitro Imaging Kit (RiboBio, China) according to the manufacturer’s instructions. Briefly, the cells were incubated with 50 μM EdU for 1 h. before fixation with 4% paraformaldehyde, permeabilization in 0.3% Triton X-100, and EdU staining. The EdU-positive cells were counted randomly in five fields under a microscope (× 100).
8
Cell Proliferation Assays: CCK-8 and EdU
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Cell Counting Kit‐8 (CCK‐8) and Ethynyl deoxyuridine (EdU) incorporation experiments were employed to determine cell proliferation ability. For CCK‐8 assay, 3000 cells were seeded per well into 96‐well plates. After culturing for indicted time, cell proliferation was evaluated using the Cell Counting Kit‐8 (Dojindo Laboratories) in accordance with the instruction. The absorbance values at 450 nm at each time point were collected to plot cell proliferation curves. EdU incorporation experiment was performed with the Cell‐Light™ EdU Apollo®643 In Vitro Imaging Kit (RiboBio, Guangzhou, China) in accordance with the instruction. The results were counted using Zeiss AxioPhot Photomicroscope (Carl Zeiss, Oberkochen, Germany) via collecting at least 5 random fields.
9
Hypoxia-Induced Cell Proliferation Assay
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HPASMCs were seeded into 96-well plates (3000 cells per well) and cultured in normoxic or hypoxic conditions as mentioned above. Cell proliferation was assessed using Edu Cell Proliferation Assay Kit (RiboBio, Cell‐Light™ Edu Apollo®643 In Vitro imaging kit), according to the manufacture’s instruction. The cells were added with 100 μL Edu (diluent reagent A with a complete medium by 1:1000) and incubated for 2 h at 37 °C. Then cells were fixed in 4% paraformaldehyde for 15–30 min and incubated with 1 × Apollo® solution for another 30 min at room temperature. Cell nuclei were stained with 100 μL 1 × Hoechst33342 for 30 min. Finally, the cells were examined under a fluorescent microscope (Olympus, Japan). Data were shown as fold-change increase in the percentage of Edu-incorporating cells in treated cells, compared with negative controls.
10
Quantifying Cell Proliferation Using EdU Assay
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SPC‐A1 cells cultured in 96‐well microplates (Cat No 655090; Greiner) were transfected with miRNA mimics or inhibitors 24 hours after seeding. At different timepoints after transfection, de novo synthesized DNA in the S phase cells were stained using the Cell‐Light EdU Apollo643 In Vitro Imaging Kit (Cat No C10310‐2; RiboBio) based on the method established by Salic et al.27 Briefly, each culture well was added with 100 μL 50 μM 5‐ethynyl‐2′‐deoxyuridine (EdU) diluted in culture media, then incubated for 2 hours and washed with phosphate‐buffered saline. Fix and permeabilize the cells with paraformaldehyde and Triton X‐100. Next, add 100 μL 1X Apollo dye to stain the replicating DNA for 30 minutes followed by nuclear staining using Hoechst 33342. After the final wash, dual channel fluorescent images of the stained cells were captured using an Operetta CLS High‐content Analysis System (PerkinElmer). Under a 10× objective, a grid of 21 fields were imaged for each well to cover nearly the whole bottom area to eliminate the bias associated with insufficient sampling. The total cell number and EdU positive cell number per well were counted by the Harmony High Content Imaging and Analysis Software (PerkinElmer). The proliferating ratio in each well was calculated as (number of EdU+cells/number of Hoechst stained cells) × 100%.
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