Axiovert 200m fluorescent

Manufactured by Zeiss

Sourced in Germany

The Axiovert 200M is a fluorescence microscope designed for advanced imaging applications. It features a sturdy, modular design and provides high-resolution imaging capabilities. The Axiovert 200M is equipped with a range of illumination options, including LED and laser sources, to support various fluorescence techniques.

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Lab products found in correlation

Axiocam hsm, by Zeiss (1 mentions) Fluoromount g medium, by Electron Microscopy Sciences (1 mentions) Lsm 510 confocal microscope, by Zeiss (1 mentions) Immersol 518f, by Zeiss (1 mentions) Axiovision software, by Zeiss (1 mentions)

Close spelling variants of this product

Axiovert 200M fluorescent microscope (56 citations) Axiovert 200M inverted fluorescent microscope (21 citations) Axiovert 200M (1358 citations) Axiovert (339 citations)

4 protocols using axiovert 200m fluorescent

Intravital Microscopy of Kidney Capillaries

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Intravital video microscopy (IVVM) was performed as described previously26 (link)30 (link). Briefly, following anesthesia with isoflurane, FITC-labeled dextran (2 μmol/kg in 3 ml/kg normal saline) was administered via the penile vein to visualize the capillary vascular space. After 10 minutes, the left kidney was exposed by a flank incision and positioned on a glass stage above an inverted Zeiss Axiovert 200M fluorescent microscope equipped with an Axiocam HSm camera (Zeiss, Germany). Videos of 10 seconds (approximately 30 frames/second) at 200X magnification were acquired from five randomly selected, non-overlapping fields of view. Body temperature was maintained at 35°–37 °C with a warming lamp. Approximately 150 capillaries were randomly selected and analyzed from these fields of view from the kidney of each animal. Capillaries were categorized as “continuous” if red blood cell movement was uninterrupted; “intermittent” if red blood cell movement stopped or reversed; or “no flow” if no red blood cell movement was observed. Data were expressed as the percentage of vessels in each of the three categories.

Li Y., Hadden C., Cooper A., Ahmed A., Wu H., Lupashin V.V., Mayeux P.R, & Kilic F. (2016). Sepsis-induced elevation in plasma serotonin facilitates endothelial hyperpermeability. Scientific Reports, 6, 22747.

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Immunofluorescence Staining Protocol

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The cells grown on coverslips in 12 well plates were fixed with 4% formaldehyde in PBS for 20 min and washed for 3 times with PBS. For regular permeabilization assays, the cells were incubated for 5 min in 0.2% Triton X100 in PBS followed by 1 h incubation in 3% membrane blocking agent (Amersham) in PBS. The same blocking solution was used for dilution of primary and secondary antibodies. For mild permeabilization assay primary and secondary antibodies were diluted in 0.02% saponin in PBS containing 5% fetal bovine serum as a blocking agent. The cells were incubated with all antibodies for one hour. Processed coverslips were mounted on slides using Fluoromount-G medium (Electron Microscopy Sciences). Images were taken using either Zeiss Axiovert 200M fluorescent or LSM 510 confocal microscope.

Viktorova E.G., Nchoutmboube J.A., Ford-Siltz L.A., Iverson E, & Belov G.A. (2018). Phospholipid synthesis fueled by lipid droplets drives the structural development of poliovirus replication organelles. PLoS Pathogens, 14(8), e1007280.

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Quantitative Colocalization Analysis

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Z-stack images for each channel were captured at ×100 magnification with Immersol 518F (Zeiss) oil immersion using a Zeiss Axiovert 200M fluorescent microscope running AxioVision software. Slices with the clearest resolution were selected for further analysis. The background was removed by subtracting the mean gray value for each channel in an area containing no cells. Colocalization analysis was performed using the ImageJ plug-in, “JaCoP”, according to the developer’s instructions. Negative controls were achieved by rotating one channel at least 90° and re-running the analysis. Positive controls were achieved by running colocalization analysis on two of the same channels. All graphs are plotted as the mean Pearson’s coefficient of at least three independent experiments with error bars representing the standard error. Each MCF-7 image contains ~15–20 cells, and MDA-MB-231 images contain ~5–10 cells per image.

Singhal S.K., Byun J.S., Park S., Yan T., Yancey R., Caban A., Hernandez S.G., Hewitt S.M., Boisvert H., Hennek S., Bobrow M., Ahmed M.S., White J., Yates C., Aukerman A., Vanguri R., Bareja R., Lenci R., Farré P.L., De Siervi A., Nápoles A.M., Vohra N, & Gardner K. (2021). Kaiso (ZBTB33) subcellular partitioning functionally links LC3A/B, the tumor microenvironment, and breast cancer survival. Communications Biology, 4, 150.

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Quantifying Transplanted Schwann Cells

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Complete sets of every 5th section (20 μm thick, 100 μm apart) of the spinal cord segments containing grafted SCs-GFP were processed. Fluorescent images were acquired with a Zeiss Axiovert 200M fluorescent microscope in Z-stack mode and were deconvoluted with the Axiovision software package (Carl Zeiss) to obtain near-confocal quality optical sections. Merged images were post-processed using ImageJ (NIH, Bethesda, MD) to quantify the total number of SCs-GFP per 20 μm-thick section. Only GFP⁺ SCs that contained a clear nuclear dye Hoechst 33342 were recognized and counted. All BrdU⁺ SCs-GFP, representing proliferating grafted SCs, were photographed and counted manually in each 20 μm-thick section. Likewise, all TUNEL⁺ SCs-GFP, representing grafted SCs that underwent apoptosis, were counted manually in each 20 μm-thick section. The estimated total number of GFP⁺ SCs was calculated as: (the number of GFP⁺ SCs per section) x (the number of total sections per set) x (the number of sets). The estimated total number of BrdU⁺ SCs-GFP and TUNEL⁺ SCs-GFP was calculated in the same manner. The percentage of BrdU⁺ or TUNEL⁺ SCs-GFP in all grafted SCs-GFP at different time points was calculated by dividing the total number of BrdU⁺ or TUNEL⁺ SCs-GFP by the total number of SCs-GFP.

Wang X, & Xu X.M. (2014). Long-term survival, axonal growth-promotion, and myelination of Schwann cells grafted into contused spinal cord in adult rats. Experimental neurology, 261, 308-319.

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