Inverted microscopy

Manufactured by Olympus

Sourced in Japan

The Inverted Microscopy is a type of optical microscope that is designed with the objective lens and the specimen positioned below the stage. This configuration allows for the observation of living cells and cell cultures, as well as other samples that require an upright orientation. The Inverted Microscopy provides a clear and detailed view of the specimen, enabling users to study its structure and behavior.

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Lab products found in correlation

Ultra low attachment plate, by Corning (3 mentions) Transwell chamber, by Corning (2 mentions) Matrigel, by BD (2 mentions) Goat serum, by Boster Bio (1 mentions) Primary rabbit anti cd31, by Abcam (1 mentions) Fv1200mpe, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions) Tunel kit, by Roche (1 mentions) Inverted microscope, by Olympus (1 mentions) Cck 8 reagent, by Wuhan Servicebio Technology (1 mentions) Image pro plus 6, by Olympus (1 mentions) Rpmi media, by Thermo Fisher Scientific (1 mentions) Schneider s insect medium, by Merck Group (1 mentions) Penicillin, by Merck Group (1 mentions) Streptomycin, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Hoechst 33258, by Merck Group (1 mentions) Inverted fluorescence microscope, by Olympus (1 mentions) 6 well plate, by Corning (1 mentions) Oil red o, by Abcam (1 mentions)

47 protocols using inverted microscopy

Oil Red O Staining of Adipocytes and Liver Tissue

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3T3-L1 adipocytes were treated with MOPEE (0, 25, 50, 100, 200, 400 μg/ml) for 24 h. Subsequently, the cells were washed twice with phosphate-buffered saline (PBS) and fixed with 4% paraformaldehyde for 30 min at room temperature. Excess 4% paraformaldehyde was removed, the cells were washed twice with PBS, and staining was performed with 60% oil red O dye (Sigma, St. Louis, MO, USA) for 10 min. Excess oil red O was removed, and the cells were then washed twice with PBS. The lipid droplets within the cells were visualized and photographed by inverted microscopy (Olympus, Tokyo, Japan).
Fresh hepatic tissues were embedded in tissue-freezing medium (Cell Path Ltd., Newtown, UK) and stored at −80°C. Subsequently, 8-μm frozen sections were fixed with 10% formaldehyde solution for 1 h at 4°C. Excess formaldehyde solution was removed, and the sections were washed three times with PBS. Staining was performed with oil red O dye for 15 min in the dark, and the sections were then rinsed with distilled water, stained with hematoxylin dye for 16 s, and rinsed again with distilled water for 5–10 min. The sections were observed and photographed by inverted microscopy (Olympus).

Xie J., Wang Y., Jiang W.W., Luo X.F., Dai T.Y., Peng L., Song S., Li L.F., Tao L., Shi C.Y., Hao R.S., Xiao R., Tian Y, & Sheng J. (2018). Moringa oleifera Leaf Petroleum Ether Extract Inhibits Lipogenesis by Activating the AMPK Signaling Pathway. Frontiers in Pharmacology, 9, 1447.

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Tumor Spheroid Formation Assay

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For tumor spheroid forming assay, CD133⁺ and CD133⁻ cells were plated (5 × 10⁴ cells/well) in ultralow attachment plates (Corning Inc.) in DMEM/F12 stem cell medium. Cells were maintained at 37°C in a humidified 5% CO₂ incubator and fed with 0.2 ml of fresh stem cell medium on days 2, 4 and 6. The total number of spheres was counted after 7 days by inverted microscopy (Olympus). To examine the self-renewal ability of CD133⁺ and CD133⁻ cells, primary spheres were dissociated with trypsin and 0.05% EDTA. Single cell suspension obtained from primary spheres were counted and replated in 6-well ultralow attachment plates. Secondary spheroids derived from single cells of primary spheres were examined by inverted microscopy (Olympus).

Kim Y., Yeon M, & Jeoung D. (2017). DDX53 Regulates Cancer Stem Cell-Like Properties by Binding to SOX-2. Molecules and Cells, 40(5), 322-330.

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Visualizing Skin Microvasculature and Thickness

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For measuring epidermal thickness, paraffin sections were stained with H and E. and images were taken by inverted microscopy (Olympus). Epidermal thickness was measured under microscope.
The immunofluorescent (IF) staining of mice skin biopsies for CD31 was performed to assess the microvessel densities. The 5‐μm thick tissues were sectioned from frozen tissue. To inhibit non‐specific antigen–antibody reactions, tissues were blocked with goat serum (Boster Biological Technology) for 30 min and washed for 3 times with PBS buffer. Primary rabbit anti‐CD31 (Abcam) was added and incubated at 4°C overnight. The next day, recovery temperature at 37°C, slides were washed in PBS for 3 times and incubated with secondary antibody (Goat anti‐rabbit IgG; Zhongshanjinqiao) for 2 h at room temperature. Finally, slices were washed for three times with PBS, incubated with 4′, 6‐diamidino‐2‐phenylindole (DAPI; Solarbio) for 10 min at room temperature. Sections were then imaged using LSCM (Olympus FV1200MPE).

Niu X., Han Q., Li X., Li J., Liu Y., Li Y., Wu Y, & Zhang K. (2022). EDIL3 influenced the αvβ3‐FAK/MEK/ERK axis of endothelial cells in psoriasis. Journal of Cellular and Molecular Medicine, 26(20), 5202-5212.

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Ocular Irritancy Evaluation of Topical Formulations

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The potential ocular irritancy and/or damaging effects of the formulations were evaluated by studying the histology of the eye tissues. Five microliter of the formulations (TA suspension, TA-liposomes, and TA-CHL) were instilled into the lower conjunctival sac of the left eye of each mouse five times a day for a period of seven days, while the right eye used as a control. At the end of the seven days, all the mice were euthanized, and the eyes were isolated and fixed in 4% paraformaldehyde. Sections were cut along sagittal sections of 3 μm per eye from the paraffin blocks and stained with hematoxylin and eosin (HE). In addition, the terminal deoxynucleotidyl transferase nick-end-labeling (TUNEL) assay was performed in tissue sections using a TUNEL kit (Roche, Switzerland). The sections were examined by inverted microscopy (Olympus, Japan). The number of TUNEL-positive cells per visual field was counted and expressed as percent of the total number of cells.

Li J., Cheng T., Tian Q., Cheng Y., Zhao L., Zhang X, & Qu Y. (2019). A more efficient ocular delivery system of triamcinolone acetonide as eye drop to the posterior segment of the eye. Drug Delivery, 26(1), 188-198.

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Histological and Immunohistochemical Analysis of Tumor Tissues

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Tumor and organ tissues were paraffin-embedded and sectioned into 4 mm after being fixed in 4% of paraformaldehyde solution for 24 h. The sections were deparaffinized and rehydrated before detection. For histological observation, tumor sections were stained with hematoxylin and eosin (HE), and then observed by an inverted microscopy (Olympus, Japan).
For immunohistochemistry analysis, the sections were treated with 3% hydrogen peroxide for 10 min to block endogenous peroxidase, and then incubated with Ki67 primary antibody for overnight at 4 °C. Subsequently, the sections were treated with secondary antibody at room temperature for 20 min. After being washed with PBS, the sections were incubated with DAB chromogenic solution for 10 min, followed by rewashing in PBS. The sections were counterstained with hematoxylin after DAB coloration. The antibody specificity was confirmed by negative control without primary antibody treatment. Immunopositive areas were observed by an inverted microscope (Olympus, Japan).

Zhang Y., Li Y., Sun C., Chen X., Han L., Wang T., Liu J., Chen X, & Zhao D. (2021). Effect of Pterostilbene, a Natural Derivative of Resveratrol, in the Treatment of Colorectal Cancer through Top1/Tdp1-Mediated DNA Repair Pathway. Cancers, 13(16), 4002.

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Investigating Osteosarcoma Cell Proliferation

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Osteosarcoma cells were trypsinized and resuspended in a complete medium. For the CCK-8 assay, cells were cultured in 96-well plates at an initial density of 3 × 10³ cells/well. CCK-8 reagent (Servicebio Technology) was added into each well at 0, 24, 48, and 72 h. After incubation for 1 h at 37 °C, the absorbance readings were obtained at 450 nm. For the colony formation assay, cells were seeded into six-well plates at an initial density of 5 × 10² cells/well and were cultured for ~10 days. Colonies were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. For the transwell invasion assay, 1 × 10⁵ cells in 200 μl serum-free medium were added into the upper well of a transwell chamber (Corning Costar, Corning, NY, USA) precoated with Matrigel, and 600 μl medium containing 20% FBS was added into the lower chamber. Invaded cells were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet and observed by inverted microscopy (Olympus, Tokyo, Japan).

Xia K., Zheng D., Wei Z., Liu W, & Guo W. (2023). TRIM26 inhibited osteosarcoma progression through destabilizing RACK1 and thus inactivation of MEK/ERK signaling. Cell Death & Disease, 14(8), 529.

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Wound Healing Assay for Cell Migration

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Migration of HaCaT and human fibroblasts was measured using the monolayer wound assay in vitro. Cells were plated in 6-well plates (1 × 10⁵ cells per well) with culture medium until completely confluent. Cells were scraped across the plate with a 200-μL pipette tip. Cells were cultured with serum-free DMEM with or without hADSC-CM and hFDSPC-CM (20 μg/mL) for 24 h. Cell migrations at 0 and 24 h were imaged with inverted microscopy (Olympus, Tokyo, Japan), and five randomly selected fields from each well were used for scratch area calculation using Image-Pro Plus 6 software. The results are presented as scratch area at 0 h − scratch area at 24 h (μm²).

Xin Y., Xu P., Wang X., Chen Y., Zhang Z, & Zhang Y. (2021). Human foreskin-derived dermal stem/progenitor cell-conditioned medium combined with hyaluronic acid promotes extracellular matrix regeneration in diabetic wounds. Stem Cell Research & Therapy, 12, 49.

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Quantifying Mature Osteoclasts via TRAP Staining

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The fixed cells and tissues were subjected to TRAP staining using the Acid Phosphatase, Leukocyte (TRAP) Kit (387A; Sigma‐Aldrich) as previously described.⁽²⁸, ³⁵⁾ TRAP‐positive cells with more than three nuclei were counted as mature osteoclasts and observed by an inverted microscopy (Olympus Corporation, Tokyo, Japan).

Zhao S., Liu H., Chen J., Qian D., Kong F., Jie J., Yin G., Li Q, & Fan J. (2020). Macrophage GIT1 Contributes to Bone Regeneration by Regulating Inflammatory Responses in an ERK/NRF2‐Dependent Way. Journal of Bone and Mineral Research, 35(10), 2015-2031.

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Modeling Acute Anti-Thy1 Mesangioproliferative Glomerulonephritis

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In this study, OX7 plus NRS was used for modeling. OX7 is an anti-thy 1 monoclonal antibody that can specifically bind to the thy-1 antigen on RMCs to form an antigen-antibody complex. NRS contain complement so that using OX7 plus NRS can simulate the pathogenesis of acute anti-thy1 MsPGN in vivo [20 (link), 21 (link)]. RMCs at passages 5 to 10 were grown in RPMI media (Gibco) with 10% fetal bovine serum (FBS). Then, the cells were placed in 6-well plates at 4.8 × 10⁵ cells/well and allowed to adhere overnight. After being washed three times with PBS, the cells were placed in serum-free media for an additional 12 h. The cells were sensitized by incubation with OX7 antibodies (0.5 μg/ml, 2.5 μg/ml and 10 μg/ml) at 37 °C for 60 min, followed by incubation with 8% NRS or serum-free RPMI. Moreover, Huaier (10 mg/ml) or an equal volume of PBS was added. RMC morphology was observed and recorded by inverted microscopy (Olympus) within 24 h.

Geng W., Tu C., Chen D., Lu Z., Mao W, & Zhu H. (2022). Huaier attenuates the adverse effects of pyroptosis by regulating the methylation of rat mesangial cells: an in vitro study. BMC Complementary Medicine and Therapies, 22, 92.

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Osteosarcoma Cell Migration Assay

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The transfected osteosarcoma cells were seeded in six-well plates and cultured to 90 % cell density. Next, a scratch was created using a 200 µL pipette tip. The scratches were observed and photographed every 24 h using inverted microscopy (Olympus, Tokyo, Japan), and the migration distance was measured.

Cheng S., Huang X, & Guo W. (2022). TIAM2 promotes proliferation and invasion of osteosarcoma cells by activating the JAK2/STAT3 signaling pathway. Journal of Bone Oncology, 37, 100461.

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