Inverted epifluorescence microscope

Manufactured by Olympus

Sourced in Japan, Germany

The Inverted epifluorescence microscope is an optical instrument designed for the study of fluorescent samples. It features a reversed orientation, where the light source and objectives are positioned below the stage, allowing for the observation of cells and other specimens in an upright position. This microscope is capable of detecting and imaging fluorescent signals emitted by labeled samples.

Automatically generated - may contain errors

Lab products found in correlation

μmanager, by Molecular Devices (2 mentions) Metamorph, by Molecular Devices (2 mentions) Hanks balanced salt solution (hbss), by Merck Group (2 mentions) Ethylene glycol tetraacetic acid (egta), by Merck Group (2 mentions) Regular hbss, by Thermo Fisher Scientific (2 mentions) Nunc lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions) Prolong gold w dapi, by Thermo Fisher Scientific (1 mentions) Cellsens software, by Olympus (1 mentions) Fluoromount gtm mounting medium, by Thermo Fisher Scientific (1 mentions) 4 6 diamidine 2 phenylindole dihydrochloride dapi, by Thermo Fisher Scientific (1 mentions) Phalloidin ifluor 555, by Abcam (1 mentions) Prolong gold antifade reagent with dapi, by Thermo Fisher Scientific (1 mentions) Dsred mito, by Takara Bio (1 mentions) Lsm 780 microscope, by Zeiss (1 mentions) Mtdsred, by Takara Bio (1 mentions) Imagej, by Olympus (1 mentions) In situ cell death detection kit, by Roche (1 mentions) Vectashield mounting medium, by Vector Laboratories (1 mentions) Alexa fluor 594 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Metamorph 7, by Molecular Devices (1 mentions)

Spelling variants of this product

25 protocols using inverted epifluorescence microscope

Histological Analysis of Oral and Skin Tissues in Grhl2 Mice

Check if the same lab product or an alternative is used in the 5 most similar protocols

Paraffin-embedded histological sections were stained with H&E for determination of the histological changes in epidermal and tongue epithelium in Grhl2 WT and KO mice, and were analyzed without blinding by an oral pathologist. Mouse oral mucosa and skin were fixed in 4% (wt/vol) paraformaldehyde at 4 °C for 24 h. Samples were embedded in paraffin, sectioned at 4 μm thickness, and stained as described previously^{7 (link)}. Numbers of positive staining cells were counted and plotted as % of all cells in at least 10 fields in each slide.
For IFS, cells were cultured in Nunc™ Lab-Tek™ II Chamber Slide™ System (ThermoFisher Scientific) to reach 70–80% confluence and fixed in 2% paraformaldehyde for 20 min. Cells were permeabilized with 0.2% Triton X-100 in phosphate-buffered saline (PBS) for 10 min, and then blocked for 1 h in PBS containing 2% FBS, and incubated overnight at 4 °C with the primary antibody. After three washes with PBS, cells were incubated with the secondary antibody for 1 h. Slides were mounted in Prolong Gold w/DAPI (Invitrogen). Images were captured on an Olympus epifluorescence inverted microscope (Olympus, Cypress, CA). IFS signals were quantified via Image J software and the corrected total cell fluorescence was calculated.

Chen W., Kang K.L., Alshaikh A., Varma S., Lin Y.L., Shin K.H., Kim R., Wang C.Y., Park N.H., Walentin K., Schmidt-Ott K.M, & Kang M.K. (2018). Grainyhead-like 2 (GRHL2) knockout abolishes oral cancer development through reciprocal regulation of the MAP kinase and TGF-β signaling pathways. Oncogenesis, 7(5), 38.

+ Open protocol

+ Expand

Immunofluorescence Analysis of hCECs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The hCECs were seeded in 384-well plates pre-coated with iMatrix-511 at a ratio of 500 cells/mm² and cultured in BGM alone (control) or BGM containing TGF-β1, 2, or 3, or one of the TGF-β cell signaling pathway inhibitors, for four weeks. The IF protocol was previously described by our team [29 (link),42 (link)]. Briefly, cells were fixed in pure methanol at room temperature for 15 min after rinsing with PBS containing Ca²⁺ and Mg²⁺. The cells were then rehydrated in PBS and incubated in blocking buffer (PBS, 2% bovine serum albumin, 2% goat serum) for 30 min at 37 °C. The primary antibodies, diluted to 1/300 in blocking buffer, were incubated with cells at 37 °C for one hour under gentle agitation (30 rpm). After three rinses in PBS, the secondary antibodies, diluted to 1/600, and DAPI diluted at 2 µg/mL in blocking buffer were incubated with cells at 37 °C for one hour under gentle agitation. After three rinses in PBS, the cells were immersed in Fluoromount-G^TM mounting medium (00-4958-02, Invitrogen) to protect the fluorochromes (Alexa Fluor™ 488 and DAPI). An epifluorescence inverted microscope (IX81, Olympus, Tokyo, Japan) with the CellSens software (Soft Imaging System GmbH, Olympus) was used to acquire images.

Aouimeur I., Sagnial T., Coulomb L., Maurin C., Thomas J., Forestier P., Ninotta S., Perrache C., Forest F., Gain P., Thuret G, & He Z. (2023). Investigating the Role of TGF-β Signaling Pathways in Human Corneal Endothelial Cell Primary Culture. Cells, 12(12), 1624.

+ Open protocol

+ Expand

Podocyte Immunofluorescence Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence experiments were performed with Phalloidin-iFluor 555 (Abcam, Cambridge, UK) and 4',6-diamidine-2'-phenylindole dihydrochloride (DAPI; Life Technologies, Carlsbad, USA) to visualise cell cytoskeleton and nucleus, respectively. Mouse anti-ZO-1 antibody (clone 1) and mouse anti-nephrin antibody (clone 174CT2.1.1), used to visualise cell-cell contact complexes, were purchased from BD Bioscience (Le Pont de Claix, France) and from Sigma-Aldrich, respectively. Anti-mouse Alexa Fluor 488 conjugated secondary antibodies, used to detect primary antibodies, were obtained from Life Technologies.
Differentiated podocytes grown on glass coverslips were fixed with paraformaldehyde (3.7% in PBS) and permeabilised with Triton X-100 (0.5%). Then the cells were incubated with primary antibodies directed against nephrin (1/50) or ZO-1 (1/50) for 1 h at room temperature, washed and incubated with the anti-mouse Alexa Fluor 488 conjugated secondary antibodies (1/200) in the presence of Phalloidin-iFluor 555 and DAPI to visualise F-actin and cell nucleus, respectively. The images were acquired using an epifluorescence inverted microscope (IX81, Olympus, Tokyo, Japan) equipped with a cell imaging software (Soft Imaging System GmbH, Munster, Germany).

Delézay O., He Z., Hodin S., Saleem M.A., Mismetti P., Perek N, & Delavenne X. (2017). Glomerular filtration drug injury: In vitro evaluation of functional and morphological podocyte perturbations. Experimental cell research, 361(2).

+ Open protocol

+ Expand

Donor-decay Kinetics Imaging Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Donor-decay time-course was studied as before (Plant et al., 2010 (link)), using an Olympus inverted epi-fluorescence microscope. Cells were studied in a solution comprising (in mM): NaCl 130, KCl 4, MgCl₂ 1.2, CaCl₂ 2, HEPES 10, pH was adjusted to 7.4 with NaOH. mTFP1 was excited at 445 nm and the emission collected through a 470–500 nm bandpass filter, YFP was excited at 514 nm and the emission collected through a 525–575 nm filter. Images were captured using a scientific camera controlled by μmanager or Metamorph software (Molecular Devices) and were analyzed with ImageJ.

Plant L.D., Xiong D., Romero J., Dai H, & Goldstein S.A. (2020). Hypoxia Produces Pro-arrhythmic Late Sodium Current in Cardiac Myocytes by SUMOylation of NaV1.5 Channels. Cell reports, 30(7), 2225-2236.e4.

+ Open protocol

+ Expand

Donor-decay Kinetics Imaging Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

Calcium Imaging of Cellular Response

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were seeded at 7 × 10⁴ cells per well in a 24-well plate. The next day, cells were pretreated with SsD for 6 h and then incubated with Fura-2 AM (4 µM) in regular HBSS (Invitrogen, 14175-059) at room temperature for 20–30 min in the dark. Subsequently, cells were washed three times with calcium-free HBSS containing 2 mM EGTA (Sigma, 3889). Finally, cells were placed on the stage of an Olympus inverted epifluorescence microscope with a ×20 objective and challenged with 200 µM GPN or 1 µM TG at room temperature for 10 min. Fluorescence images were collected by excitation alternating between 340 nm and 380 nm with emission at 510 nm. Images were captured every 5 s and processed by Cell R imaging software.

Li C., Huang L., Sun W., Chen Y., He M.L., Yue J, & Ballard H. (2019). Saikosaponin D suppresses enterovirus A71 infection by inhibiting autophagy. Signal Transduction and Targeted Therapy, 4, 4.

+ Open protocol

+ Expand

ROS Detection in LPA-Treated Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For ROS measurement by DCF fluorescence, cells were pre-incubated with serum-containing media (and, where indicated, 400 U/mL PEG-catalase) overnight, then treated or not with 1 μM VPC32183, 1 μM DPI, or 500 nM apocynin for 30 min prior to incubation with or without 100 nM alkyl-LPA (18:1) for 30 min. Cells were incubated with 50 μM DCFH diacetate for the final 10 min of LPA treatment, then washed and visualized using an Olympus inverted epi-fluorescence microscope with FITC filters.

Klomsiri C., Rogers L.C., Soito L., McCauley A.K., King S.B., Nelson K.J., Poole L.B, & Daniel L.W. (2014). Endosomal H2O2 production leads to localized cysteine sulfenic acid formation on proteins during lysophosphatidic acid-mediated cell signaling. Free radical biology & medicine, 71, 49-60.

+ Open protocol

+ Expand

Immunohistochemistry and Western blotting protocols

Cited 3 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Basic IHC and IF procedures in FFPE sections (4 μm) were detailed previously^{11 (link),14 (link)–17 (link)}. For IHC, slides were deparaffinized in xylene and hydrated in gradient alcohols to water. Slides were treated with antigen retrieval in 10 mM citrate buffer (pH 6.0) and incubated with primary antibodies (Supplementary Table 1; optimal dilutions were determined by antibody titration experiments in pilot studies) followed by secondary antibodies and DAPI counterstaining (when applicable). IHC images were captured using an Olympus inverted (epifluorescence) microscope. The whole-mount and TMA IHC slides were scanned using Aperio ScanScope imaging system (Aperio Technologies, Vista, CA, USA) and a ×40 objective. Images were analyzed using the ScanScope software. For Western blotting analysis, whole cell lysate was prepared in RIPA buffer and run on 4–15% gradient SDS-PAGE gels. The proteins were transferred to nitrocellulose membrane followed by incubation with primary antibodies (Supplementary Table 1) and corresponding secondary antibodies. Films were developed using Western Lighting ECL Plus reagent (PerkinElmer). Representative original films for multiple Western blotting analyses were presented in Supplementary Figs. 20–25.

Li Q., Deng Q., Chao H.P., Liu X., Lu Y., Lin K., Liu B., Tang G.W., Zhang D., Tracz A., Jeter C., Rycaj K., Calhoun-Davis T., Huang J., Rubin M.A., Beltran H., Shen J., Chatta G., Puzanov I., Mohler J.L., Wang J., Zhao R., Kirk J., Chen X, & Tang D.G. (2018). Linking prostate cancer cell AR heterogeneity to distinct castration and enzalutamide responses. Nature Communications, 9, 3600.

+ Open protocol

+ Expand

Clonal and Clonogenic Assays for AR-KO and AR+ LNCaP Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For in vitro clonal assays, 5000 AR-KO or AR⁺ LNCaP cells were plated in 6-well plates, and 2 weeks later, cells were stained with 1:20 Giemsa and images scanned with an HP scanner. The number of clones was quantified with ImageJ. For clonogenic assays, 500 AR-KO or AR⁺ cells were suspended in 100 μL 50% Matrigel and 50% normal media containing 0.1 nM DHT or 2 μM Enza. The single cell suspension was spread on the edge of wells in a 24-well plate. 2 weeks later, images were taken with an Olympus inverted epifluorescence microscope and sphere numbers were counted.

+ Open protocol

+ Expand

Measuring Intracellular Calcium Dynamics

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were cultured in 24-well plates at the density of 7 × 10⁴ cells/well in regular medium overnight and were labeled with 4 μM Fura-2 AM in regular HBSS (Invitrogen, 14175-059) at room temperature for 30 min. The cells were then washed with calcium-free HBSS containing 2 mM EGTA (Sigma, 3889) 3 times and incubated in the presence or absence of vacuolin-1 at room temperature for another 10 min. Cells were put on the stage of an Olympus inverted epifluorescence microscope (Telefon, Germany) and visualized using a 20× objective. Fluorescence images were obtained by alternate excitation at 340 nm and 380 nm with emission set at 510 nm. Images were collected by a CCD camera every 3 or 6 s and analyzed by Cell R imaging software.

Lu Y., Dong S., Hao B., Li C., Zhu K., Guo W., Wang Q., Cheung K.H., Wong C.W., Wu W.T., Markus H, & Yue J. (2014). Vacuolin-1 potently and reversibly inhibits autophagosome-lysosome fusion by activating RAB5A. Autophagy, 10(11), 1895-1905.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!