Axioplan 2 imaging microscope

Manufactured by Zeiss

Sourced in Germany, United States, France

The Axioplan 2 imaging microscope is a high-performance research microscope designed for advanced imaging applications. It features a modular design, allowing for customization to meet specific research needs. The microscope provides excellent optical performance, with a wide range of magnification and illumination options. It is suitable for a variety of sample types and is commonly used in fields such as biology, materials science, and medical research.

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Spelling variants of this product

Axioplan 2 microscope (979 citations) Axioplan 2 (859 citations) Axioplan microscope (451 citations) Axioplan (304 citations) Axioplan 2 imaging (143 citations) Axioplan 2 imaging fluorescence microscope (14 citations) Axioplan 2 imaging fluorescent microscope (6 citations) Axioplan imaging microscope (3 citations) Axioplan 2 Imaging light microscope (2 citations) AxioPlan 2 imaging widefield microscope (2 citations)

184 protocols using axioplan 2 imaging microscope

Imaging Hippocampal Neurons and Microglia

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Hippocampal neuron morphology within the pyramidal shaped CA1 neurons were imaged (z-stack thickness of 0.5 μm) using an Axioplan 2 imaging microscope (Zeiss) equipped with a 63× (N.A. 1) oil objective accompanied with a digital camera (AxioCam MRm, Zeiss).
The microscopic images of anti–IBA-1 and Aβ (clone BAM-10) were taken within the area of cortex and hippocampus. IBA-1 cells were taken from the CA1 area of the hippocampus in three-dimensional (z-stack thickness, 1 µm) using Axioplan 2 imaging microscope (Zeiss) equipped with an ApoTome module (Zeiss) with a 20× objective (NA, 0.8) and a digital camera (AxioCam MRm; Zeiss). To analyze, a region of interest was drawn in ImageJ software (Wayne Rasband, NIH, Bethesda, MD).

Lonnemann N., Hosseini S., Marchetti C., Skouras D.B., Stefanoni D., D’Alessandro A., Dinarello C.A, & Korte M. (2020). The NLRP3 inflammasome inhibitor OLT1177 rescues cognitive impairment in a mouse model of Alzheimer’s disease. Proceedings of the National Academy of Sciences of the United States of America, 117(50), 32145-32154.

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Cell Counting Using Stereological Methods

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To estimate the number of the labeled cells, cell counts were made using the serial sections, as described previously (Dokter et al., 2015 (link)). In brief: countings were performed according to the Abercrombie’s correction formula (starting around Bregma −1.06 mm), since this method renders biases within the range of the optical disector by taking into account that the particles counted are small compared with the section thickness (von Bartheld, 2002 (link)). No guard zones were used, since the use of guard zones can bias even optical disector counting (Baryshnikova et al., 2006 (link)). The Linderstrom-Lang/Abercrombie (LLA) equation for estimating numerical neuronal densities is:

N = n * t (t + H) or N / n = f = t / (t + H)

N is an estimate of the number of objects in the defined region, n is the counted number of objects, t is the mean thickness of the virtual section, H is the mean height of the objects, and f is the conversion factor for converting n to N.
In a first step, n was quantified using an Axioplan 2 imaging microscope (Zeiss, Germany) fitted for fluorescence. In a second step, H, the height of the cells in the z-axis, was estimated using a computer-driven motorized stage (Merzhäuser, Germany) connected to the Axioplan 2 imaging microscope (Zeiss, Germany) under the control of StereoInvestigator (MBF Biosciences, USA).

Poser R., Dokter M., von Bohlen und Halbach V., Berger S.M., Busch R., Baldus M., Unsicker K, & von Bohlen und Halbach O. (2015). Impact of a deletion of the full-length and short isoform of p75NTR on cholinergic innervation and the population of postmitotic doublecortin positive cells in the dentate gyrus. Frontiers in Neuroanatomy, 9, 63.

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Polysaccharide Capsule and Melanin Production

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Formation of polysaccharide capsule was examined by differential interference microscopy (DIC) on an Axioplan 2 imaging microscope (Zeiss), after incubation for 24 h at 30°C in defined low-iron medium (LIM) and staining with India ink. Melanin production was examined on L-3,4-dihydroxyphenylalanine (L-DOPA) plates containing 0.1% glucose (Tangen et al., 2007 (link)). The influence of copper on melanin production was investigated on L-DOPA plates supplemented with 100 μM or 1 mM of CuSO₄. To observe vacuole morphology, cells from overnight cultures were incubated with lipophilic vacuole dye MDY-64 (Invitrogen, USA, 2.5 μM final concentration) for 15 minutes on ice and washed with liquid YPD medium. Cells were incubated at 30°C in YPD for an additional 30 minutes before visualized under fluorescence and DIC microscopy on an Axioplan 2 imaging microscope (Zeiss) with magnification 1000X, and a Zen Lite software. An additional vacuole-sequestered dye, carboxy-DCFDA (5-(and-6)-carboxy-2’,7’-dichlorofluorescein diacetate, Invitrogen, USA, at 10 μM final concentration), was used to assess vacuole morphology (Harrison et al., 2002 (link)).

Hu G., Bakkeren E., Caza M., Horianopoulos L., Sánchez-León E., Sorensen M., Jung W, & Kronstad J.W. (2021). Vam6/Vps39/TRAP1-domain proteins influence vacuolar morphology, iron acquisition and virulence in Cryptococcus neoformans. Cellular microbiology.

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Subcellular Localization and Lamellipodia Visualization

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To observe Ct-OATP1B3 subcellular localization, cells were incubated with primary antibodies against Ct-OATP1B3 (1:100, Santa Cruz, USA) at 4 °C overnight, then incubated with secondary antibody at 37 °C for 1 h and co-stained with DAPI for 5 min at room temperature. To observe cellular lamellipodia, cells were incubated with anti-cortactin antibody (1:100, Bioss, China) for overnight at 4 °C, then treated with 100 nM TRITC Phalloidin (Solarbio, China) for 30 min. Cell micrographs were obtained using a Zeiss Axioplan 2 imaging microscope (Zeiss AG, Oberkochen, Germany).

Huang Y., Du Y., Zheng Y., Wen C., Zou H., Huang J., Zhou H., Zhao H, & Wu L. (2022). Ct-OATP1B3 promotes high-grade serous ovarian cancer metastasis by regulation of fatty acid beta-oxidation and oxidative phosphorylation. Cell Death & Disease, 13(6), 556.

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Lentiviral Transduction of mCherry-eGFP-LC3B

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Lentiviral vectors (pMD2GVSGV, pMDlg/pRRE, and pRSV‐rev) are used to pack and deliver mCherry‐eGFP‐LC3B and a puromycin resistance gene to H358 and A549 as described (Parejo et al, 2019 (link)). Selection in puromycin is conducted for 3–5 days starting 2 days post‐infection. Images were acquired on a ZEISS Axioplan 2 imaging microscope (Carl Zeiss MicroImaging, Göttingen, Germany) and processed using Adobe Photoshop CS6 v.13 (Adobe Systems, San Jose, CA, USA). Rationmetrics of mCherry/eGFP was performed on a BD Biosciences LSRII flow cytometer.

Yang Z., Liang S., Saliakoura M., Yang H., Vassella E., Konstantinidou G., Tschan M., Hegedüs B., Zhao L., Gao Y., Xu D., Deng H., Marti T.M., Kocher G.J., Wang W., Schmid R.A, & Peng R. (2021). Synergistic effects of FGFR1 and PLK1 inhibitors target a metabolic liability in KRAS‐mutant cancer. EMBO Molecular Medicine, 13(9), e13193.

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Microscopy Imaging and Analysis

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All immunofluorescent photographs were acquired using a Zeiss Axioplan 2 Imaging microscope (Carl Zeiss Microimaging, Inc., Thornwood, NY, USA) with a digital black and white camera. Images were analyzed using the Zeiss Axiovision Image Analysis software v4.6 (Carl Zeiss Microimaging, Inc.).

Mac Nair C.E., Fernandes K.A., Schlamp C.L., Libby R.T, & Nickells R.W. (2014). Tumor necrosis factor alpha has an early protective effect on retinal ganglion cells after optic nerve crush. Journal of Neuroinflammation, 11, 194.

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Histopathological Analysis of Murine Lungs

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The chemotherapeutic murine model was performed additionally for histopathology. Three mice in each group (including the CEA10, ΔtslA, and ΔtslA+tslA strain groups) were humanely euthanized at day 2 and day 4 postinoculation. Lungs were harvested from each group and fixed in 10% formalin before embedding in paraffin was performed. Sections (5 μm in thickness) were taken and stained with either H&E (hematoxylin and eosin) or GMS (Gomori’s methenamine silver stain) as previously described (72 (link)). Slides were analyzed microscopically with a Zeiss Axioplan 2 imaging microscope (Carl Zeiss Microimaging, Inc., Thornwood, NY) fitted with a QImaging Retiga-SRV Fast 1394 red-green-blue (RGB) camera. The analysis was performed in Phylum Live 4 imaging software. Images were captured at ×50 magnification as indicated in each image.

Thammahong A., Caffrey-Card A.K., Dhingra S., Obar J.J, & Cramer R.A. (2017). Aspergillus fumigatus Trehalose-Regulatory Subunit Homolog Moonlights To Mediate Cell Wall Homeostasis through Modulation of Chitin Synthase Activity. mBio, 8(2), e00056-17.

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Murine Model of Aspergillus Infection

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CD-1 female mice, 20–24 grams, were immune suppressed as described above. Mice were inoculated with 2x10⁶ conidia of CEA10, ΔcreA, or creA^R and lungs were harvested 48 hours post inoculation for histopatholoical sectioning and staining. For early germination studies, immune suppressed mice were inoculated with 1x10⁷ conidia and lungs were harvested 8 hpi for sectioning and staining. Briefly, lungs were perfused with 10% buffered formalin solution upon collection, then fixed in 10% buffered formalin overnight. Lungs were blocked in paraffin, sectioned, and stained with Gömöri methenamine silver (GMS) and hematoxylin and eosin (H&E) stains. Images were obtained with a Zeiss Axioplan 2 imaging microscope (Carl Zeiss Microimaging, Inc.) fitted with Qimiging RETIGA-SRV Fast 1394 RGB camera using Phylum Live 4 imaging software.

Beattie S.R., Mark K.M., Thammahong A., Ries L.N., Dhingra S., Caffrey-Carr A.K., Cheng C., Black C.C., Bowyer P., Bromley M.J., Obar J.J., Goldman G.H, & Cramer R.A. (2017). Filamentous fungal carbon catabolite repression supports metabolic plasticity and stress responses essential for disease progression. PLoS Pathogens, 13(4), e1006340.

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Immunohistochemical Staining of CD8+ T Cells

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The snap frozen tumor tissue were sectioned (thickness: 4 μm) and stained with rat anti-mouse CD8 antibody (1:200) from Abcam (Cambridge, UK). Tissue slides were blocked with 0.1% bovine serum albumin for 1 hour before incubation with primary antibodies overnight at 4 °C. After incubation, the slides were washed 3 times with PBS and then incubated with fluorochrome conjugated secondary antibodies (1:400) for one hour at room temperature. Stained slides were washed 3 times with PBS and mounted with ProLong® Antifade Mountant solution containing 4’, 6-diamidino-2-phenylindole (DAPI) (Invitrogen, Carlsbad, CA). All stained slides were stored in dark at 4 °C, and examined within 3 days using a Zeiss Axioplan 2 imaging microscope (Carl Zeiss, Hamburg, Germany). Representative images were captured using a Spot digital camera (Diagnostic Instruments Inc., Sterling Heights, Michigan, USA). The original magnifications were ×200.

He L., Law P.T., Wong C.K., Chan J.C, & Chan P.K. (2017). Exendin-4 Exhibits Enhanced Anti-tumor Effects in Diabetic Mice. Scientific Reports, 7, 1791.

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Immunofluorescence Analysis of Spleen Sections

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Spleens were snap-frozen in OCT compound (Sakura Finetek, Torrance, CA) at the time of sacrifice. Cryostat spleen sections (5 μm) were fixed in acetone, washed with PBS, and blocked with PBS/5% fetal bovine serum. Spleen sections were stained with: FITC-anti-IgD or -anti-CD45.1/CD45.2 (BD Biosciences); biotin-conjugated PNA (Sigma-Aldrich), HEL, anti-CD45.1/CD45.2, anti-B220 (BD Biosciences), anti-IgM^a, or anti-CD4 (Cedarlane Laboratories); and PE-anti-IgM^a or -anti-CD4 (BD Biosciences). Biotinylated Ab staining was revealed with rhodamine (tetramethylrhodamine)-conjugated streptavidin (Molecular Probes) or 7-amino-4-methylcoumarin-3-acetic acid-conjugated streptavidin (AMCA, Jackson ImmunoResearch) as a secondary reagent. Stained sections were mounted with Fluoro-Gel (Electron Microscopy Sciences), and tissue fluorescence was visualized using a Zeiss Axioplan 2 imaging microscope (Zeiss, Oberkochen, Germany).

Chang N.H., Manion K.P., Loh C., Pau E., Baglaenko Y, & Wither J.E. (2017). Multiple tolerance defects contribute to the breach of B cell tolerance in New Zealand Black chromosome 1 congenic mice. PLoS ONE, 12(6), e0179506.

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