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Axioplan 2 microscope system

Manufactured by Zeiss
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Sourced in Germany

The Axioplan 2 Microscope System is a high-performance optical microscope designed for advanced imaging and analysis applications. It features a modular design that allows for the integration of various components and accessories to meet the specific needs of the user. The Axioplan 2 provides reliable and consistent performance for a wide range of microscopy techniques, including brightfield, darkfield, phase contrast, and differential interference contrast (DIC) imaging.

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Lab products found in correlation

Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (4 mentions) Triton x 100, by Merck Group (3 mentions) Metamorph analysis software, by Zeiss (3 mentions) Alexa fluor 488 donkey anti goat igg antibody, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Positively charged slides, by Thermo Fisher Scientific (2 mentions) Normal goat serum (ngs), by Cell Signaling Technology (2 mentions) H6908, by Merck Group (2 mentions) Alexa fluor 488 goat anti mouse igg antibody, by Thermo Fisher Scientific (1 mentions) Bmk 2202, by Vector Laboratories (1 mentions) Ab8592, by Abcam (1 mentions) Bf f3, by Leibniz Institute DSMZ (1 mentions) Alizarin red s, by Merck Group (1 mentions) A11055, by Thermo Fisher Scientific (1 mentions) Sigma kit ht15, by Merck Group (1 mentions) Histochoice clearing agent, by Merck Group (1 mentions) Histomount, by National Diagnostics (1 mentions) Eosin y solution, by Merck Group (1 mentions) Cryostat, by Leica (1 mentions) Alexa fluor 594 donkey anti rabbit igg antibody, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Axioplan 2 microscope (979 citations) Axioplan 2 (859 citations) Axioplan microscope (451 citations) Axioplan (304 citations) AxioPlan 2 IE Motorized Microscope System (2 citations) Axioplan 2 fluorescent microscope system (2 citations)

26 protocols using axioplan 2 microscope system

1

Histochemical Analysis of Plant Sections

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Fresh hand-cut sections were subjected to histochemical analysis (described below) and photographed through with a Zeiss AxioPlan 2 microscope system with automatic exposure. Wiesner staining was performed by incubating sections in 1% phloroglucinol in ethanol:water (7:3) with 30% HCl. Mäule staining was performed by first incubating sections in KMnO4 (1%). After 10 min, sections were washed and acidified with HCl (30%) for 1 min, washed again, and then incubated in NaHCO3 (5%).
Voxeur A., Soubigou-Taconnat L., Legée F., Sakai K., Antelme S., Durand-Tardif M., Lapierre C, & Sibout R. (2017). Altered lignification in mur1-1 a mutant deficient in GDP-L-fucose synthesis with reduced RG-II cross linking. PLoS ONE, 12(9), e0184820.
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2

Cardiac Utrophin, β-DG, and β-SG Immunofluorescence

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For assessment of utrophin, fresh frozen cardiac cross-sections (10 μm thick) were blocked for 30 min with 10% fetal bovine serum (FBS)/PBS and then incubated with primary antibodies in 5% FBS and PBS for 2 hr at room temperature. For assessment of β-DG and β-SG, a mouse on mouse (MOM) detection kit (BMK-2202; Vector Laboratories; USA) was used per manufacturer’s instructions. Primary antibodies and dilutions used were as follows: utrophin A (developed in-house as previously described14 (link)) 1:2,000, β-SG (Novocastra; NCL-L-b-SARC) 1:50, and β-DG (Novocastra; NCL-b-DG) 1:100. Sections assessed for utrophin were reacted with Alexa Fluor 488 donkey anti-goat IgG antibody (Thermo Fisher Scientific; A-11055) 1:2,000 for 1 hr at room temperature. Sections assessed for β-SG and β-DG were reacted with Alexa Fluor 488 goat anti-mouse IgG antibody (Life Technologies; ab150117) 1:100 for 1 hr at room temperature. All sections were rinsed in PBS before air drying and application of coverslip with fluorescent mounting medium. Intensity was detected using a fluorescence microscope (Axioplan 2 Microscope System; Carl Zeiss, Germany). Utrophin staining, imaging, and image selection were performed blind.
Kennedy T.L., Guiraud S., Edwards B., Squire S., Moir L., Babbs A., Odom G., Golebiowski D., Schneider J., Chamberlain J.S, & Davies K.E. (2018). Micro-utrophin Improves Cardiac and Skeletal Muscle Function of Severely Affected D2/mdx Mice. Molecular Therapy. Methods & Clinical Development, 11, 92-105.
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3

Muscle Fiber Quantification via Laminin-α2/MyHC-emb Staining

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Co-staining Laminin-α2/MyHC-emb were performed on transverse QUAD and EDL sections as previously described and images obtained with the Axioplan 2 Microscope System (Carl Zeiss). Laminin-α2 is used as a mask to obtain total number of muscle fibers per image. Four 10× images per muscle were quantified per muscle and mouse and percent MyHC-emb positivity obtained by dividing the number of muscle fiber positive for MYH3 by total number of fibers. All counting was performed blinded.
Guiraud S., Edwards B., Squire S.E., Moir L., Berg A., Babbs A., Ramadan N., Wood M.J, & Davies K.E. (2018). Embryonic myosin is a regeneration marker to monitor utrophin-based therapies for DMD. Human Molecular Genetics, 28(2), 307-319.
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4

Muscle Cross-Section Immunofluorescence

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Fresh frozen muscle cross-sections were blocked for 30 min with 10% FBS/PBS and then incubated with Alexa Fluor ® 488 goat anti-mouse IgG (Life Technologies; ab150117) antibody, 1:750 in 5% FBS/PBS overnight at 4 °C. Sections were rinsed in PBS before air drying and application of cover with fluorescent mounting medium, intensity was detected using a fluorescence microscope (Axioplan 2 Microscope System; Carl Zeiss, Germany).
Kennedy T.L., Moir L., Hemming S., Edwards B., Squire S., Davies K, & Guiraud S. (2017). Utrophin influences mitochondrial pathology and oxidative stress in dystrophic muscle. Skeletal Muscle, 7, 22.
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5

Multimodal Muscle Imaging Techniques

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Frozen transverse and longitudinal muscle sections were blocked in 10% foetal bovine serum/PBS for 20 min, incubated with primary antibodies for 1 h at room temperature or overnight at 4°C, washed in PBS and incubated with suitable Alexa Fluor secondary antibodies for 1 h at room temperature. Sections were examined under an Axioplan 2 Microscope System (Carl Zeiss, Germany). The following antibodies and dilution were used: goat polyclonal anti-utrophin (1:500, URD40) (75 (link)), mouse monoclonal anti- β-dystroglycan (1:100, MANDAG2, CIND), rabbit monoclonal anti- α1-CTFP/dystrobrevin (1:1000) (76 (link)), rabbit polyclonal anti desmin (1:100, ab8592, abcam), rabbit polyconal anti-collagen type I (1:100, 600-401-103-0.5, Rockland), mouse anti-MYHC1 (1:200, B8-F8), mouse anti-MYHC2A (1:200; SC-71); mouse IgM anti-MYHC2B (1:100, BF-F3; all from German Collection of Microorganisms and Cell Cultures) and polyclonal anti- α-Bungarotoxin AlexaFluo 488 conjugate antibody (1:500, B-13422, Life Technologies).
Guiraud S., Squire S.E., Edwards B., Chen H., Burns D.T., Shah N., Babbs A., Davies S.G., Wynne G.M., Russell A.J., Elsey D., Wilson F.X., Tinsley J.M, & Davies K.E. (2015). Second-generation compound for the modulation of utrophin in the therapy of DMD. Human Molecular Genetics, 24(15), 4212-4224.
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6

Histological Assessment of Muscle and Cardiac Tissue

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Serial cross-sections (10 μm) were cut through the hearts, diaphragm, EDL, soleus, and TA muscles using a refrigerated (−20°C) cryostat (Bright OTF5000 Cryostat, Casa Alvarez Scientific Material, Spain). To assess muscle architecture, fresh-frozen sections were reacted with H&E. Calcification was assessed by alizarin red Staining, sections were incubated with 2% alizarin red S (Sigma-Aldrich [pH 4.2]) for 1–5 min and washed with distilled (d)H2O and cleared. To assess fibrosis in the heart, cardiac cross-sections were stained with Masson’s trichrome as described previously.49 (link) All sections were air-dried before application of coverslip and mounting medium and acquisition of digital images (Axioplan 2 Microscope System; Carl Zeiss, Germany). Quantified of alizarin red and Masson’s trichrome was performed using Fiji ImageJ 1.49i software and expressed as percentage area.
Kennedy T.L., Guiraud S., Edwards B., Squire S., Moir L., Babbs A., Odom G., Golebiowski D., Schneider J., Chamberlain J.S, & Davies K.E. (2018). Micro-utrophin Improves Cardiac and Skeletal Muscle Function of Severely Affected D2/mdx Mice. Molecular Therapy. Methods & Clinical Development, 11, 92-105.
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7

Utrophin Immunohistochemistry in TA Muscle

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Frozen TA muscle sections (10 μm thick) were fixed in acetone for 10 min, washed for 5 min in PBS and blocked for 1 h in MOM blocking solution (M.O.M.™ Kit, FMK-2202. Vector Laboratories). Following 2 × 2 min washes, sections were incubated overnight with primary antibodies at 4 °C. The following antibody and dilutions were used: goat polyclonal anti-utrophin (1:500, URD40). After 2 × 2 min washes and 5 min in MOM diluent (M.O.M.™ Kit, FMK-2202. Vector Laboratories), sections were incubated in anti-goat (1:2000; A11055, Life Technologies) Alexa Fluor® 488 secondary antibody for 2 h at room temperature. Sections were examined under an Axioplan 2 Microscope System (Carl Zeiss, Germany) and multi-acquisition module used to obtain images.
Babbs A., Berg A., Chatzopoulou M., Davies K.E., Davies S.G., Edwards B., Elsey D.J., Emer E., Figuccia A.L., Fletcher A.M., Guiraud S., Harriman S., Moir L., Robinson N., Rowley J.A., Russell A.J., Squire S.E., Thomson J.E., Tinsley J.M., Wilson F.X, & Wynne G.M. (2020). Synthesis of SMT022357 enantiomers and in vivo evaluation in a Duchenne muscular dystrophy mouse model. Tetrahedron, 76(2), 130819.
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8

Histological Analysis of Skeletal Muscle

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Muscle samples were frozen in liquid nitrogen-chilled isopentane, and stored at −80°C. Transverse and longitudinal extensor digitorum longus (EDL), tibialis anterior (TA), soleus (SOL), diaphragm (DIA) and heart cryosections (10 μm) were stained with Haematoxylin and Eosin solution (H&E), Masson's trichrome (MT) and Alizarin red (AR). The Axioplan 2 Microscope System (Carl Zeiss, Germany) was used to obtain the images. The proportion of centrally nucleated fibres was determined by analysing the H&E images. Areas of necrosis were quantified based on the DMD_M.1.2.007 MDC1A_M.1.2.004 TREAT-NMD SOPS and performed with the Fiji ImageJ 1.49i software (73 ) on the whole EDL, SOL and DIA sections. Fibrosis of the whole DIA sections was observed using Masson's trichrome staining to visualize connective tissue and muscle fibres (Sigma kit HT15; SigmaAldrich) and quantified using a previously published macro (74 (link)) with the Fiji ImageJ 1.49i software.
Guiraud S., Squire S.E., Edwards B., Chen H., Burns D.T., Shah N., Babbs A., Davies S.G., Wynne G.M., Russell A.J., Elsey D., Wilson F.X., Tinsley J.M, & Davies K.E. (2015). Second-generation compound for the modulation of utrophin in the therapy of DMD. Human Molecular Genetics, 24(15), 4212-4224.
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9

Histological Staining of Frozen Muscle

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Frozen TA muscle sections (10 μm thick) were air dried for 10 min before being placed in hematoxylin solution (Sigma, UK #GHS232) for 8 min, slides were washed in tap water to remove excess stain and placed in 70% ethanol, 1% HCl for 10 s followed by 5 s in 1% Eosin Y solution (Sigma, UK #E4382) in 80% ethanol. Following a series of alcohol washes unbound stain was removed in Histochoice clearing agent (Sigma UK #H2779) and mounted using Histomount (National Diagnostics, UK #HS-103). Slides were examined under an Axioplan 2 Microscope System (Carl Zeiss, Germany) to obtain images.
Babbs A., Berg A., Chatzopoulou M., Davies K.E., Davies S.G., Edwards B., Elsey D.J., Emer E., Figuccia A.L., Fletcher A.M., Guiraud S., Harriman S., Moir L., Robinson N., Rowley J.A., Russell A.J., Squire S.E., Thomson J.E., Tinsley J.M., Wilson F.X, & Wynne G.M. (2020). Synthesis of SMT022357 enantiomers and in vivo evaluation in a Duchenne muscular dystrophy mouse model. Tetrahedron, 76(2), 130819.
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10

Immunofluorescent Muscle Fiber Analysis

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Fresh-frozen muscle cross-sections were blocked for 30 min with 10% FBS and PBS and then incubated with Alexa Fluor 488 goat anti-mouse IgG (Life Technologies; ab150117) antibody, 1:750 in 5% FBS and PBS overnight at 4°C. Sections were rinsed in PBS before air drying and application of cover with fluorescent mounting medium; intensity was detected using a fluorescence microscope (Axioplan 2 Microscope System; Carl Zeiss, Germany).
Kennedy T.L., Guiraud S., Edwards B., Squire S., Moir L., Babbs A., Odom G., Golebiowski D., Schneider J., Chamberlain J.S, & Davies K.E. (2018). Micro-utrophin Improves Cardiac and Skeletal Muscle Function of Severely Affected D2/mdx Mice. Molecular Therapy. Methods & Clinical Development, 11, 92-105.
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