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Inverted microscope

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The Inverted Microscope is a type of optical microscope where the objective lens and the specimen are positioned below the stage. This design allows for the observation of living cells and samples in their natural state. The Inverted Microscope provides a clear, high-resolution image of the specimen, enabling detailed analysis and examination.

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Lab products found in correlation

1010 electron microscope, by JEOL (2 mentions) Rpmi glutamax, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by GE Healthcare (1 mentions) Novolink polymer detection system, by Leica (1 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions) Hepes, by Thermo Fisher Scientific (1 mentions) Ht 29, by American Type Culture Collection (1 mentions) Heat inactivated fetal calf serum, by GE Healthcare (1 mentions) Abt 737, by Selleck Chemicals (1 mentions) Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions) Af488 conjugated goat anti mouse igg, by BioLegend (1 mentions) Ab55152, by Abcam (1 mentions) Igg2a, by BioLegend (1 mentions) 96 well round bottom ultra low attachment microplates, by Corning (1 mentions) Alizarin red solution, by Merck Group (1 mentions) Hybrid cell count bz h2c software, by Keyence (1 mentions) Zetasizer nano z, by Malvern Panalytical (1 mentions) Su8220, by Hitachi (1 mentions) Toc l, by Shimadzu (1 mentions)

Spelling variants of this product

Biorevo BZ-9000 inverted microscope (8 citations) BZ-9000 inverted fluorescence microscope (8 citations) BZ-X710 inverted microscope (7 citations) BZ-X710 inverted fluorescence microscope (4 citations) Inverted optical microscope (4 citations) BZ-9000 inverted fluorescence/bright field microscope (3 citations) BZ-X700 inverted microscope (3 citations) Inverted fluorescence phase contrast microscope BZ-X710 (2 citations) BZ-X710 inverted fluorescent microscope (2 citations) Inverted fluorescent microscope (2 citations)

9 protocols using inverted microscope

1

3D CRC Cell Culture and Apoptosis Assay

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The human CRC cell line HT29 was purchased from ATCC (Manassas, VA, USA). Cells were maintained in RPMI+GlutaMAX (Gibco, Karlsruhe, Germany) supplemented with 10% heat-inactivated fetal calf serum (PAA Laboratories, Colbe, Germany), 1% penicillin/streptomycin (PAA Laboratories), 1% HEPES (Gibco) and 1% non-essential amino acids (Gibco) and cultured in a humidified atmosphere (37 °C, 5% CO2). The cells were regularly tested for contaminations and routinely subcultured twice a week.
To grow human HT29 CRC cells in a three-dimensional culture assay, Alvetex scaffolds (Reinnervate, Sedgefield, UK) were used. Seeding of cells was done as described previously.11 (link) After 24 h, medium was changed and additionally supplemented with 1 μM ABT-737 (Selleckchem, Munich, Germany) or DMSO as control. After 4 days of treatment, in which medium and drug were renewed every second day, scaffolds were harvested, cryosectioned and immunostained as described.11 (link) For staining, a primary antibody against cleaved PARP (Abcam) and the NovoLink Polymer Detection System (Leica Microsystems) have been used, according to the manufacturer's protocol. At least 10 pictures per section were captured with an inverted microscope (Keyence, Neu-Isenburg, Germany), and the number of cleaved PARP-positive cells was determined by manual counting.
Scherr A.L., Gdynia G., Salou M., Radhakrishnan P., Duglova K., Heller A., Keim S., Kautz N., Jassowicz A., Elssner C., He Y.W., Jaeger D., Heikenwalder M., Schneider M., Weber A., Roth W., Schulze-Bergkamen H, & Koehler B.C. (2016). Bcl-xL is an oncogenic driver in colorectal cancer. Cell Death & Disease, 7(8), e2342-.
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2

Immunofluorescent Staining of Adipose Tissue

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Fresh tissue sections were obtained from the subcutaneous obese AT, they were placed in O.C.T. blocks (Tissue-Tek 4583) and stored at −80°C. Blocks were sectioned at 16 μm thickness and fixed in cold acetone for 10 min. For the staining, the tissue sections were fixed in acetone and washed twice with PBS. The tissue sections were then blocked for 1 h with 5% BSA at room temperature, and incubated with primary anti-CD1d antibody (abcam ab11076), or with primary anti-MHC class II antibody (abcam ab55152), both 1:100 diluted, overnight at 4°C. The next day, the slides were washed with PBS twice for 5 min and incubated with the following secondary antibodies: AF488-conjugated goat anti-mouse IgG (Biolegend 405319) for CD1d detection, and AF647-conjugated goat anti-mouse IgG (ThermoFisher InVitrogen A-21236) for MHC class II detection, both 1:100 diluted, 1 h at room temperature, or with the isotype controls [IgG1 (Biolegend 401402) and IgG2a (Biolegend 401502), respectively]. The slides where then washed 3 times, 3 min each. The cover slides were mounted with ProLong Gold Antifade Mountant and DAPI (4′,6-diamidino-2-phenylindole, ThermoFisher Scientific), which stains the nuclei of immune cells, to visualize the nuclei. Slides were imaged with a Keyence inverted microscope.
Frasca D., Diaz A., Romero M., Garcia D., Jayram D., Thaller S., del Carmen Piqueras M., Bhattacharya S, & Blomberg B.B. (2020). Identification and Characterization of Adipose Tissue-Derived Human Antibodies With “Anti-self” Specificity. Frontiers in Immunology, 11, 392.
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3

Huh-7 and Hep3B Spheroid Formation

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Huh‐7 and Hep3B cells were seeded into 96‐well round bottom ultra‐low attachment microplates (Corning, NY, USA) at densities of 1000 and 2500 cells, respectively, in 200 μL of growth medium supplemented with 10% FBS per well. The cells were allowed to spontaneously aggregate, as described previously.24 Spheroids were observed with an inverted microscope (Keyence, Osaka, Japan) and the sectional area [S (×10 000 μm2)] of each spheroid was measured with Image J software (NIH, Bethesda, MD, USA). The volume index of the spheroid was calculated as S1.5.
Kasai Y., Toriguchi K., Hatano E., Nishi K., Ohno M., Yoh T., Fukuyama K., Nishio T., Okuno M., Iwaisako K., Seo S., Taura K., Kurokawa M., Kunichika M., Uemoto S, & Nishi E. (2017). Nardilysin promotes hepatocellular carcinoma through activation of signal transducer and activator of transcription 3. Cancer Science, 108(5), 910-917.
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4

Alizarin Red Staining of Calcium Nodules

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After different treatment strategies, MC3T3-E1 cells were cultured in ODIM for 14 days, followed by being fixed with 10% paraformaldehyde for 10 min. Then, cells were stained with 1% alizarin red solution (Sigma-Aldrich, California, USA) for 5 min. Lastly, the inverted microscope (KEYENCE, Tokyo, Japan) was utilized to take images of the calcium nodules [23 (link)].
Wang Q., Xie X., Zhang D., Mao F., Wang S, & Liao Y. (2022). Saxagliptin enhances osteogenic differentiation in MC3T3-E1 cells, dependent on the activation of AMP-activated protein kinase α (AMPKα)/runt-related transcription factor-2 (Runx-2). Bioengineered, 13(1), 431-439.
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5

Quantification of Aortic Immune Cells

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Histologic images were analyzed using an inverted microscope (KEYENCE), and quantification was performed using the Hybrid cell count BZ-H2C software (KEYENCE). The number of macrophages (CD68+ cells) was determined by counting all immune-stained cell counts per aortic cross section. The levels of HIF-1α, MMPs, and pimonidazole were evaluated using immunohistochemical staining, and staining images were analyzed using the Image J software (National Institutes of Health, Bethesda, Md). We evaluated three cross-sections from each tissue sample taken at 100-μm intervals and calculated the mean. We set a threshold to compute for the area positive for each protein and then calculated the ratio of the positive area to the total aortic area.
Unno N., Tanaka H., Yata T., Kayama T., Yamanaka Y., Tsuyuki H., Sano M., Inuzuka K., Naruse E, & Takeuchi H. (2020). K-134, a phosphodiesterase 3 inhibitor, reduces vascular inflammation and hypoxia, and prevents rupture of experimental abdominal aortic aneurysms. JVS-Vascular Science, 1, 219-232.
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6

Comprehensive Characterization of Surface Wettability

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A field emission scanning electron microscope was used to examine the surface morphology of the samples as they were developed (SEM, HITACHI-SU8220, Hitachi, Ltd., Tokyo, Japan). Chemical component was characterized by energy dispersive spectrometry (EDS, Bruker, Billerica, MA, USA). Data-physics was used to characterize surface wettability, including water contact angle (WCA), oil contact angle (OCA), underoil–water contact angle (UWCA), and underwater–oil contact angle (UOCA) (Data-Physics, DataPhysics Instrumente GmbH., Filderstadt, Germany). The liquid droplets utilized for WCA, UWCA, CA and UOCA analyses had a volume of 4 μL. The amount of water remained in the collected filtrates was determined by Karl Fischer Titrator (TP653, TimePower Measure and Control Equipment Co. Ltd., Beijing, China). Total organic carbon (TOC) equipment was used to measure the oil content of the filtrates (TOC-L, Shimadzu (Shanghai) Global Laboratory Consumables Co. Ltd., Shanghai, China). The Zetasizer Nano ZS (Malvern Instruments Ltd., Malvern, UK) was used to investigate the dispersion of water droplets in oil and oil droplets in water. The microscopy pictures of W/O and O/W emulsions were acquired by using an inverted microscope (Keyence Corporation, Osaka, Japan).
Liu X., Feng S., Wang C., Yan D., Chen L, & Wang B. (2022). Wettability Improvement in Oil–Water Separation by Nano-Pillar ZnO Texturing. Nanomaterials, 12(5), 740.
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7

Adipocyte and Muscle Fiber Analysis

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eWAT and iWAT depots were fixed in 4 % paraformaldehyde overnight, dehydrated in ethanol, embedded in paraffin, and sectioned. Sections were stained with hematoxylin and eosin. Images were taken at 10x magnification on a Keyence inverted microscope. The Adiposoft plugin for ImageJ was used to quantify adipocyte area [41 (link)]. Four animals per group and two images per animal were quantified (n = 8 per group) and average adipocyte size was calculated. TA muscles were fixed in 10 % buffered formalin overnight, dehydrated in ethanol, embedded in paraffin, and sectioned. Sections were stained with hematoxylin and eosin. Images were taken at 20x magnification on a Keyence inverted microscope. Individual muscle fibers were manually selected and cross-sectional area was quantified using ImageJ. Two animals per group and two images per animal were quantified (n = 4 per group) and average fiber cross-sectional area was calculated.
Nunn E., Jaiswal N., Gavin M., Uehara K., Stefkovich M., Drareni K., Calhoun R., Lee M., Holman C.D., Baur J.A., Seale P, & Titchenell P.M. (2024). Antibody blockade of activin type II receptors preserves skeletal muscle mass and enhances fat loss during GLP-1 receptor agonism. Molecular Metabolism, 80, 101880.
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8

Histology and TEM Analysis of BAT

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Tissues were fixed in 4% PFA overnight, washed in PBS, dehydrated in ethanol, paraffin-embedded and sectioned. Sections were stained with hematoxylin and eosin. Images were captured on a Keyence inverted microscope. For transmission electron microscopy, BAT was fixed in 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 M sodium cacodylate buffer (pH 7.4) overnight at 4°C. Thin sections were stained with uranyl acetate and lead citrate and analyzed with a JEOL 1010 electron microscope.
Angueira A.R., Shapira S.N., Ishibashi J., Sampat S., Sostre-Colón J., Emmett M.J., Titchenell P.M., Lazar M.A., Lim H.W, & Seale P. (2020). Early B Cell Factor Activity Controls Developmental and Adaptive Thermogenic Gene Programming in Adipocytes. Cell reports, 30(9), 2869-2878.e4.
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9

Histology and TEM Analysis of BAT

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Tissues were fixed in 4% PFA overnight, washed in PBS, dehydrated in ethanol, paraffin-embedded and sectioned. Sections were stained with hematoxylin and eosin. Images were captured on a Keyence inverted microscope. For transmission electron microscopy, BAT was fixed in 2.5% glutaraldehyde, 2.0% paraformaldehyde in 0.1 M sodium cacodylate buffer (pH 7.4) overnight at 4°C. Thin sections were stained with uranyl acetate and lead citrate and analyzed with a JEOL 1010 electron microscope.
Angueira A.R., Shapira S.N., Ishibashi J., Sampat S., Sostre-Colón J., Emmett M.J., Titchenell P.M., Lazar M.A., Lim H.W, & Seale P. (2020). Early B Cell Factor Activity Controls Developmental and Adaptive Thermogenic Gene Programming in Adipocytes. Cell reports, 30(9), 2869-2878.e4.
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