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Inverted uorescence microscope

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The Inverted Fluorescence Microscope is a laboratory instrument designed to observe and analyze specimens using fluorescence imaging techniques. It features an inverted configuration, where the light source and objectives are positioned below the stage, allowing for the observation of cells and other samples in their natural, adherent state. The microscope is equipped with specialized filters and optics to detect and capture the emission of fluorescent signals from stained or labeled specimens.

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Lab products found in correlation

Dapi fluoromount g, by Southern Biotech (3 mentions) Cck 8 assay, by Dojindo Laboratories (2 mentions) Elx800 reader, by Agilent Technologies (2 mentions) Matrigel matrix, by Corning (2 mentions) Crystal violet, by Beyotime (2 mentions) Transwell chamber, by Corning (2 mentions) Matrigel matrix, by BD (2 mentions) R conjugated donkey anti rabbit igg, by Santa Cruz Biotechnology (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Dual luciferase reporter kit, by Promega (1 mentions) E cadherin, by Proteintech (1 mentions) Vimentin, by Proteintech (1 mentions) Alexa fluor 597 conjugated secondary antibody, by ZSGB-BIO (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

Close spelling variants of this product

Leica DMi-8 inverted epi uorescence microscope Uorescence microscope Inverted microscope

8 protocols using inverted uorescence microscope

1

Evaluating HCC Cell Viability and Motility

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The CCK-8 assay (Dojindo Japan) was performed as previously described [12] , human HCC cells (1×10 4 ) were seeded in the 96-well plate and incubated with ATL (0, 10, 100 or 500 μM) for 24 hours, 48 hours and 72 hours. Absorbance was recorded at 450 nm using Elx800 Reader (Bio-Tek Instruments Inc., Winooski, VT, USA).
Migration and invasion assays.
Cells (2 x 10 4 ) migration was analyzed using the transwell chamber (8 μ pore size; Corning Incorporated, Corning, NY, USA) without the Matrigel matrix. Cells (2 x 10 4 ) were seeded into the upper chamber precoated with Matrigel matrix (BD Biosciences) for invasion analysis. After incubation for 24 h, cells in the down chamber were stained with 0.1% crystal violet (Beyotime) and photographed by an inverted uorescence microscope (Leica Microsystems GmbH, Wetzlar, Germany).
Sheng L., Li J., Li N., Gong L., Liu L., Zhang Q., Li X., Luo H., & Chen Z. (2020). Atractylenolide III predisposes miR-195-5p/FGFR1 signaling axis to exert tumor-suppressive functions in liver cancer.
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2

Immunofluorescence Cytoskeleton and Adhesion Assay

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Cells were slightly washed by preheated (37°C) PBS 3 times and xed in 4% paraformaldehyde for 20 min. Then, the cells were permeated with 0.5% Triton X-100 for 5 min and blocked in 5% BSA for 1h at room temperature. For FITC-phalloidine cytoskeleton staining, F-actin was stained with TRITC (SolarBio, Beijing, China) containing 1% BSA for 40 min at room temperature. For cell IF staining, the cells were incubated with the rabbit polyclonal anti-E-cadherin antibody (diluted 1:100) and the rabbit polyclonal anti-Vimentin antibody (diluted 1:100) primary antibodies at 4°C overnight. The next day, the relevant secondary antibodies were added to the above cells for 1h at room temperature. The nuclei were stained with DAPI for 5 ~ 8min. The cells were imaged using an inverted uorescence microscope (Leica Microsystems Inc., USA).
TOP/FOP ash reporter assay A549 and H1299 with stable RBM10 overexpression were cultured in 24-well plates (2 × 10 4 cells per well). After 24h, cells were transfected with the TOP-Flash or FOP-Flash reporter plasmids together with pRL-TK using Lipofectamine 2000 (Invitrogen). After 48h of culture, the luciferase activity was analyzed using a dual-luciferase reporter kit (Promega). Data are presented as the ratio of relative light units of TOP ash to FOP ash from triplicate experiments.
Cao Y., Wang X., Meng Q., Geng J., Xu S., Lang Y., Zhou Y., Qi L., Wang Z., Wei Z., Yu Y., & Pan B. (2021). RBM10 Interacts with CTBNNBIP1 and Represses Lung Adenocarcinoma Progression Through the Wnt/β- catenin Pathway.
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3

Evaluating HCC Cell Viability and Motility

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The CCK-8 assay (Dojindo Japan) was performed as previously described [12] , human HCC cells (1×10 4 ) were seeded in the 96-well plate and incubated with ATL (0, 10, 100 or 500 μM) for 24 hours, 48 hours and 72 hours. Absorbance was recorded at 450 nm using Elx800 Reader (Bio-Tek Instruments Inc., Winooski, VT, USA).
Migration and invasion assays.
Cells (2 x 10 4 ) migration was analyzed using the transwell chamber (8 μ pore size; Corning Incorporated, Corning, NY, USA) without the Matrigel matrix. Cells (2 x 10 4 ) were seeded into the upper chamber precoated with Matrigel matrix (BD Biosciences) for invasion analysis. After incubation for 24 h, cells in the down chamber were stained with 0.1% crystal violet (Beyotime) and photographed by an inverted uorescence microscope (Leica Microsystems GmbH, Wetzlar, Germany).
Sheng L., Li J., Li N., Gong L., Liu L., Zhang Q., Li X., Luo H, & Chen Z. (2021). Atractylenolide III predisposes miR-195-5p/FGFR1 signaling axis to exert tumor-suppressive functions in liver cancer. Journal of food biochemistry, 45(5).
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4

Immunofluorescence Staining of SiHa and HeLa Cells

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SiHa and HeLa cells were incubated with primary antibodies overnight at 4 °C. After washing three times, the cells were incubated with R-conjugated Donkey anti-rabbit IgG (1:200 dilution, sc-2095, Santa Cruz Biotechnology, USA) and FITC-conjugated Donkey anti-goat IgG (1:200 dilution, EK033, Zhuangzhibio, China) for 1 h at room temperature. Finally, DAPI Fluoromount-G (SouthernBiotech, USA) was used to counterstain the cell nuclei. The uorescent was detected, and images were taken by Leica inverted uorescence microscope. The primary antibodies are listed in table 2.
Zhang Q., Tian S., Zhao M., Yang T., Quan S., Yang Q., Song L., & Yang X. (2020). SUV39H1-DNMT3A-mediated epigenetic regulation of Tim-3 and galectin-9 in the cervical cancer.
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5

Immunofluorescence Staining of SiHa and HeLa Cells

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SiHa and HeLa cells were incubated with primary antibodies overnight at 4 °C. After washing three times, the cells were incubated with R-conjugated Donkey anti-rabbit IgG (1:200 dilution, sc-2095, Santa Cruz Biotechnology, USA) and FITC-conjugated Donkey anti-goat IgG (1:200 dilution, EK033, Zhuangzhibio, China) for 1 h at room temperature. Finally, DAPI Fluoromount-G (SouthernBiotech, USA) was used to counterstain the cell nuclei. The uorescent was detected, and images were taken by Leica inverted uorescence microscope. The primary antibodies are listed in table 2.
Zhang Q., Tian S., Zhao M., Yang T., Quan S., Yang Q., Song L., & Yang X. (2020). SUV39H1-mediated DNMT3A is participated in the epigenetic regulation of Tim-3 and galectin-9 in cervical cancer.
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6

Immunofluorescence Staining of SiHa and HeLa Cells

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SiHa and HeLa cells were incubated with the primary antibody overnight at 4 °C. After thorough washing, the cells were incubated with Cy3-conjugated goat anti-rabbit IgG and FITC-conjugated Donkey anti-goat IgG for 1 h at room temperature. Finally, DAPI Fluoromount-G (SouthernBiotech) was used to counterstain the cell nuclei. The uorescent was detected, and images were taken by Leica inverted uorescence microscope. The antibodies are listed in table 2.
Zhang L., Tian S., Zhao M., Yang T., Quan S., Yang Q., Song L., & Yang X. (2020). SUV39H1-mediated DNMT3A is participated in the epigenetic regulation of Tim-3 and galectin-9 in cervical cancer.
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7

Cell Cycle and Proliferation Assay

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Glioma cells were and stained with Propidium iodide (PI) in the presence of RNase A for 15 min. Flow cytometer (BD Biosciences) was used to perform cell cycle analysis according to the protocol.
Ethynyl-2'-deoxyuridine (EdU) cell proliferation assay EdU assay kit (Ribobio, China) was used to test the cell proliferation ability according to the manufacturer's instructions. Glioma cells were seeded into wells of poly-l-ornithine precoated 12-well plates. Cells were then incubated with 200 μl of 5-ethynyl-20-deoxyuridine for 2 h at 37℃. Nuclei were counterstained with Hoechst 33342. Representative images were obtained with a Leica inverted uorescence microscope.
Xue H., Qi Y., Jin C., Qiu W., Zhao R., Wang S., Li B., Zhang Z., Guo Q., Zhang S., Gao Z., Zhao S., Pan Z., Fan Y., Chen Z., Wang H., Xu J., Deng L., Ni S., Wang J., Xue F., & Li G. (2021). The Double Face of Glioma Exosomal microRNAs: Glioma Eliminates Tumor Suppressor miR-1298-5p via Exosomes to Promote Immunosuppressive Effects of MDSCs.
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8

Immunofluorescence Staining of Cell Markers

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The cells were seeded on the poly-L-lysine (ZSGB-Bio, China) coated coverslips and then xed post culturing. After xation by adopting 4% paraformaldehyde solution at room temperature, the cells were washed with 0.01M PBS solution for 3 times. The cells was added 5% goat serum blocking liquid with 0.3% Triton X-100 for 30 minutes. The IF was performed in the wet box by overnight incubation at 4 ℃ with primary antibodies against CHN1(1:150, Abcam, UK), E-cadherin (1:200, Proteintech Group, USA), and vimentin (1:300, Proteintech Group, USA), respectively. The Alexa Fluor® 597-conjugated secondary antibody (1:300, ZSGB-Bio, China) was incubated at 37 ℃ for 1 h after the 0.01M PBS washing. The cell nuclei were stained with DAPI (0.2 µg/ml, Sigma, USA) and images were acquired by inverted uorescence microscope (Leica, Germany).
Zhao H., Wang L., Wang S., Chen X., Liang M., Zhang X., Wang J., & Xu X. (2021). CHN1 Promotes EMT via the Akt / GSK-3β / Snail Pathway in Cervical Carcinoma.
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