Fluorescence microscope

Manufactured by Kodak

The Kodak Fluorescence Microscope is an optical microscope that uses fluorescence to produce high-contrast images of samples. It utilizes a high-intensity light source, such as a mercury or LED lamp, to excite fluorescent molecules within the specimen, which then emit light at a different wavelength that is captured by the microscope's optical system and detected by a camera or eyepiece.

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Lab products found in correlation

Digital camera, by Kodak (3 mentions) Ecl kit, by GE Healthcare (1 mentions) 8 chamber slide, by Thermo Fisher Scientific (1 mentions) Formalin, by Merck Group (1 mentions) Vectashield, by Vector Laboratories (1 mentions)

4 protocols using fluorescence microscope

Western Blot and Immunofluorescence Analysis

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Cells were lysed in buffer containing 50 mM HEPES, 150 mN NaCl (4.38 g), 1 mM EDTA, 1% (w/v) CHAPS and Sigma protease inhibitor cocktail. Subsequently the cell lysates were resolved by10% SDS-PAGE and transferred to nitrocellulose membrane. The following antibodies were used: GFP antibody (632381) was purchased from Cell Signaling Technology, Bcl-x antibody (610211) was purchased from Clontech. α-Tubulin antibodies (T5168) were purchased from Sigma-Aldrich. Bound antibodies were visualized with the ECL kit (GE Healthcare).
Two hundred ninety-three cells stably transfected with circular RNA expression vector upon were seeded onto poly-lysine coated glass coverslips in a 6-well plate, and induced with 1 μg/mL tetracycline. At 48 h after induction, the cells were fixed with 4% formaldehyde in 1× PBS for 20 min, washed three times with 1× PBS, and then permeabilized with 0.2% Triton X-100. The cover slips were then mounted with mounting medium (Vector shield's mounting medium with DAPI). Cells were visualized using an Olympus fluorescence microscope, and photographs were generated using a Kodak digital camera.

Wang Y, & Wang Z. (2015). Efficient backsplicing produces translatable circular mRNAs. RNA, 21(2), 172-179.

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Localization of TEAD4 and YAP by Immunofluorescence

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To determine the localization of TEAD4-FL, TEAD4-S and YAP, we performed immunofluorescence assay. In brief, cells were plated on coverslips to appropriate density. Transfected cells were fixed on the coverslips with 4% paraformaldehyde in 1 × PBS for 15 min at room temperature and washed with 1 × PBS three times. Cells were then permeabilized with 0.2% Triton X-100 for 10 min. After blocking in 3% bovine serum albumin for 30 min, slides were incubated with indicated antibodies (Flag or Myc, 1:100 dilution) antibody diluted in 1% bovine serum albumin for 2 h. Subsequently, slides were washed with 1 × PBS for three times, and then incubated with fluorophore-conjugated secondary antibodies for 1 h. The coverslips were then washed and mounted with mounting medium (Vector shield's mounting medium with 4,6-diamidino-2-phenylindole). Cells were visualized using an Olympus fluorescence microscope, and photographs were generated using a Kodak digital camera.

Qi Y., Yu J., Han W., Fan X., Qian H., Wei H., Tsai Y.H., Zhao J., Zhang W., Liu Q., Meng S., Wang Y, & Wang Z. (2016). A splicing isoform of TEAD4 attenuates the Hippo–YAP signalling to inhibit tumour proliferation. Nature Communications, 7, ncomms11840.

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Immunofluorescence Assay for Mucin Expression

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Cells were plated on 8-chamber slides (Thermo Fisher Scientific) at approximately 2 × 10⁴ cells/chamber and incubated overnight at 37°C. Following removal of the medium, cells were fixed in 4% formalin (Sigma-Aldrich) for 15 min at RT, and then treated with 0.1% Triton X-100 in PBS for another 15 min. After washed twice with PBS, cells were incubated with 10 μg/ml of hPAM4 and a murine mAb against MUC5AC, α-MUC1, or a rabbit polyclonal antibody against MUC17 in PBS plus 1% BSA for 45 min at RT. Afterwards, cells were washed twice and incubated with a mixture of FITC-GAH and Cy3-GAM or Cy3-GAR in PBS plus 1% BSA for 30 min at RT. After three washes, chambers were dissembled. Slides were mounted with an antifade solution (VectaShield, Vector Laboratories) containing the nuclear counterstain, 4, 6-diamidino-2-phenylindole (DAPI). Image acquisition and analyses were performed using an Olympus fluorescence microscope with a Kodak camera system.

Liu D., Chang C.H., Gold D.V, & Goldenberg D.M. (2015). Identification of PAM4 (clivatuzumab)-reactive epitope on MUC5AC: A promising biomarker and therapeutic target for pancreatic cancer. Oncotarget, 6(6), 4274-4285.

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Immunofluorescence Staining of Transfected Cells

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Transfected cells with appropriate density were fixed on the coverslips with 4% paraformaldehyde in 1× PBS for 15 min at room temperature and washed with 1× PBS three times. Cells were then permeabilized with 0.2% Triton X-100 for 10 min and washed with 1 × PBS three times. After blocking in 3% BSA for 30 min, slides were incubated with indicated antibodies (p-H2AX, SRSF1, or HA, 1:100 dilution) antibody diluted in 3% BSA for 2 h. Subsequently, slides were rinsed three times with 1× PBS for 5 min each and then incubated with fluorophore-conjugated secondary antibodies for 1 h. The cover slips were then washed with 1× PBS for three times and mounted with mounting medium (Vector shield's mounting medium with DAPI). Cells were visualized using an Olympus fluorescence microscope, and photographs were captured using a Kodak digital camera.

Sheng J., Zhao Q., Zhao J., Zhang W., Sun Y., Qin P., Lv Y., Bai L., Yang Q., Chen L., Qi Y., Zhang G., Zhang L., Gu C., Deng X., Liu H., Meng S., Gu H., Liu Q., Coulson J.M., Li X., Sun B, & Wang Y. (2018). SRSF1 modulates PTPMT1 alternative splicing to regulate lung cancer cell radioresistance. EBioMedicine, 38, 113-126.

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