Cell proliferation and drug sensitivity were tested using the MTT assay. Exponentially growing cells were seeded into 96-well plates and cultured overnight to allow for cell adherence. Cell viability was measured using MTT (5 mg/ml) (Sigma-Aldrich, MO, USA). EdU incorporation was performed using an Apollo567 in vitro imaging kit (RiboBio Co., Ltd, Guangzhou, China) according to the manufacturer’s protocol. Cells were incubated with 10 μM EdU for 2 h, then fixed with 4% paraformaldehyde. After permeabilization with 0.3% Triton X-100, cells were stained with Apollo fluorescent dyes and 5 μg/ml DAPI.
Apollo567 in vitro imaging kit
Manufactured by RiboBio
Sourced in China
The Apollo567 In Vitro Imaging Kit is a laboratory instrument designed for high-resolution imaging of cellular and subcellular structures. It utilizes advanced fluorescence microscopy techniques to capture detailed images of samples in a controlled in vitro environment.
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18 protocols using apollo567 in vitro imaging kit
1
MTT Assay for Cell Proliferation
2
EdU Incorporation Assay
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EdU incorporation analysis was performed using the Apollo567 In Vitro Imaging Kit (RiboBio Inc.) according to the manufacturer’s protocol. Cells were incubated with 10 μM EdU for 2 h before fixation with 4% paraformaldehyde, permeabilization with 0.3% Triton X-100, and staining with the Apollo fluorescent dyes. Cell nuclei were stained with 5 μg/ml DAPI (4’,6-diamidino-2-phenylindole) for 10 min. The number of EdU-positive cells were counted under a microscope in five random fields. All assays were independently performed at least three times.
3
EdU Incorporation Assay with Apollo567
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An Apollo567 in vitro Imaging Kit was purchased from RiboBio Corporation (Guangzhou, China) and was applied for the EdU incorporation assay. After culturing with EdU (10 μM) for 2 h, the cells were fixed with paraformaldehyde (4%), permeabilized with Triton X-100 (0.2%), and contained with 4′,6-diamidino-2-phenylindole (DAPI, 5 μg/mL) and Apollo fluorescent dyes.
4
EdU Incorporation Imaging Protocol
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EdU incorporation was assessed using an Apollo567 In Vitro Imaging Kit (RiboBio Co., Ltd., Guangzhou, China) according to the manufacturer's protocol. Cells were incubated with 10 μM EdU for 2 h and then fixed with 4% paraformaldehyde. After permeabilization with 0.3% Triton X-100, the cells were stained with Apollo fluorescent dyes and 5 μg/ml DAPI.
5
EdU Incorporation Assay in Myoblasts
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The Cell-Light TM EdU (5-ethynyl-2’-deoxyuridine) Apollo®567 In Vitro Imaging Kit was purchased from RiboBio (Guangzhou, China). Myoblasts at the normal growth stage were incubated with 50 µM EdU culture medium for 2 h, which was prepared according to the kit instructions. Then, the cells that were labelled with EdU were dyed in Apollo reaction solution, and the cell nuclei were stained with Hoechst for 10 min. Then, the cells were observed by using an inverted fluorescence microscope (OLYMPUS, CKX53), and the data were analysed with Image J.
6
Colony Formation and Cell Proliferation Assay
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For the colony formation assay, cells were plated in 6-well plates at a density of 200 cells/well. The medium was refreshed after 24 h of incubation. Colonies were cultured for 14 days, then fixed with methanol for 10 min, and stained with a hematoxylin solution. The number of colonies containing>50 cells was counted under a microscope (Olympus, Tokyo, Japan).
EdU incorporation assay was performed using the Apollo567
in vitro imaging kit (Guangzhou RiboBio) according to the manufacturer’s protocol. In brief, cells were incubated with 10 mM EdU for 2 h before being fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, and stained with Apollo fluorescent dyes (Sigma-Aldrich). The cell nuclei were stained with 5 mg/mL of 4′,6-diamidino-2-phenylindole (DAPI) for 10 min. The number of EdU-positive cells in five random fields was counted under a microscope. All assays were independently performed in triplicate.
EdU incorporation assay was performed using the Apollo567
in vitro imaging kit (Guangzhou RiboBio) according to the manufacturer’s protocol. In brief, cells were incubated with 10 mM EdU for 2 h before being fixed with 4% paraformaldehyde, permeabilized with 0.3% Triton X-100, and stained with Apollo fluorescent dyes (Sigma-Aldrich). The cell nuclei were stained with 5 mg/mL of 4′,6-diamidino-2-phenylindole (DAPI) for 10 min. The number of EdU-positive cells in five random fields was counted under a microscope. All assays were independently performed in triplicate.
7
EdU Incorporation Analysis of Cultured Cells
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According to the manufacturer's instructions, EdU incorporation analysis was carried out through the Apollo 567 In Vitro Imaging Kit purchased from RiboBio Corporation (Guangzhou, China). After co‐culture with EdU solution at a concentration of 1/1000 for 2 h, the cells were fixed with paraformaldehyde (4%) for 30 min, permeated with Triton Xmi 100 (0.5%), and stained with 1x Apollo and 4‐diamino‐2‐phenylindole (DAPI).
8
EdU Incorporation and Cell Staining
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EdU incorporation was performed by using Apollo567 In Vitro Imaging Kit (RiboBio, Guangzhou, China) according to the manufacturer's protocol. Cells were incubated with 10 μM EdU for 2 h before fixation with 4% paraformaldehyde. After permeabilization with 0.3% Triton X-100, the cells were stained with Apollo fluorescent dyes. Cell nuclei were stained with DAPI.
9
Cell Proliferation and Apoptosis Assays
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Cell proliferation was measured by the CCK8 assay. Cells were plated into 96-well plates, and cell viability was detected by the CCK8 reagent purchased from the APExBLO company. Cells at a density of 2000 cells/well were plated in 6-well plates. After culturing for 14 days, the generated colonies were fixed with methanol for 30 min, stained with a hematoxylin solution, and photographed using a microscope. An Apollo567 in vitro Imaging Kit was purchased from RiboBio Corporation (Guangzhou, China) and was applied for the EdU incorporation assay. The apoptosis kit for TRIB3-overexpressed cells’ apoptosis detection is from Solarbio (CA1020). Annexin V and propidium iodide (PI) were used for labeling to assess cell apoptosis.
10
EdU Incorporation Assay Protocol
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Apollo567 in vitro Imaging Kit (RiboBio Co., Ltd, Guangzhou, China) was used for EdU incorporation assay, following the manufacturer's instructions. The cells were incubated with EDU reagent (10 µM) for 2 hours, fixed in 4% paraformaldehyde. The cells were labeled using Apollo fluorescent dye and 5 µg/ml DAPI after permeabilization using 0.3% Triton X-100.
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