Z1 axio observer apotome

Manufactured by Zeiss

Sourced in Germany

The Z1 Axio Observer Apotome is a microscope system designed for advanced fluorescence imaging. It features a structured illumination technique that enhances optical sectioning capabilities, enabling high-quality imaging of thick samples. The core function of this product is to provide researchers with a tool for detailed analysis of biological specimens.

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Lab products found in correlation

Vglut1, by Synaptic Systems (3 mentions) Ab13970, by Santa Cruz Biotechnology (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Mouse anti nestin, by Abcam (2 mentions) Rabbit anti tom20, by Santa Cruz Biotechnology (2 mentions) Mouse anti ssea4, by Cell Signaling Technology (2 mentions) Mouse anti tra 1 60, by Cell Signaling Technology (2 mentions) Mouse anti tra 1 81, by Cell Signaling Technology (2 mentions) Alexa fluor 555, by Thermo Fisher Scientific (2 mentions) Alexa fluor 647, by Thermo Fisher Scientific (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (2 mentions) Rabbit anti pax6, by Fortrea (2 mentions) Mitosox red, by Thermo Fisher Scientific (1 mentions) Mitotracker green, by Thermo Fisher Scientific (1 mentions) Ad nc, by GenePharma (1 mentions) Phospho histone h2a x, by Cell Signaling Technology (1 mentions) Alexa fluor 488 goat anti rabbit, by Abcam (1 mentions) Zen software, by Zeiss (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)

Close spelling variants of this product

Axio Observer Z1 with Apotome Axio Observer z1 Apotome fluorescence microscope Apotome-equipped Axio Observer Z1 microscope Axio Observer Z1 Inverted Microscope with Apotome Apotome Axio Observer Z1 inverted microscope Axio Observer Apotome Axio Observer Z1 Observer Z1 Axio Observer Apotome

21 protocols using z1 axio observer apotome

Visualizing Mitochondrial ROS and Morphology

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For detecting mitochondrial ROS and morphology, cells were incubated with 0.5 mL of HEPES medium containing 0.5 µM Mitotracker Green (Molecular Probes, Eugene, OR, USA) and 1.5 µM Mitosox Red (Molecular Probes, Eugene, OR, USA) for 20 min at 37 °C. Cells were washed thrice with 1 mL of warm HEPES buffer. Once washed, cells were imaged immediately at 6–8 random places under Axio Observer Z1Apotome (Zeiss, Jena, Germany) equipped with an incubation system to maintain 5% CO₂ and 37 °C at 490/516 nm (green) and 510/580 nm (red). Networks of mitochondria were analyzed using Mitochondrial Network Analysis (MiNA) toolset macros used along with ImageJ [65 (link)]. Mean fluorescence intensity of mitochondrial ROS was calculated using ImageJ. Around 40–50 cells were counted for analysis.

Bola S., Subramanian P., Calzia D., Dahl A., Panfoli I., Funk R.H, & Roehlecke C. (2023). Analysis of Electric Field Stimulation in Blue Light Stressed 661W Cells. International Journal of Molecular Sciences, 24(4), 3433.

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Overexpression of uc.333 miRNA

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Adenoviral vectors overexpressing uc.333 mimic (Ad-uc.333) and control vector (Ad-NC) were constructed by Genepharma (Shanghai, China). The adenovirus vectors were transfected into HepG2 cells at the density of 100 multiple of infection (MOI). Transfection efficiency was detected under a Zeiss Axio Observer Z1 Apotome fluorescence microscope.

Zhang Y., Sun J., Yao H., Lin Y., Wei J., Hu G., Guo J, & Li J. (2020). Ultraconserved element uc.333 increases insulin sensitivity by binding to miR-223. Aging (Albany NY), 12(8), 6667-6679.

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Immunofluorescent Detection of DNA Damage

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Cells were rinsed briefly for 1 min with 2 mL PBS and fixed with 4% PFA for 5 min at room temperature. Once fixed, cells were gently washed with 2 mL PBS and permeabilized with 0.5 mL of 100% ice cold methanol for 10 min at −20 °C. Once rinsed with 2 mL of PBS for 1 min, cells were blocked with 2% BSA/PBS for 30 min at room temperature. Subsequently, 0.25 mL of rabbit polyclonal Phospho-Histone H2AX (1:150; Cell Signaling, Danvers, MA, USA) primary antibody was added to the cells and incubated for overnight at 4 °C. Following day cells were washed thrice with 2 mL PBS for 5 min each and incubated with 0.25 mL of goat anti rabbit Alexa Fluor 488 (1:800, Abcam, Cambridge, UK) secondary antibody at room temperature for 1 h in the dark. Cells were rinsed thrice with 2 mL PBS for 5 min each and incubated with DAPI (1:2500, Sigma Aldrich, Schnelldorf, Germany) for 10 min. Finally, cells were mounted with 2.5% DABCO (in PBS/Glycerol) solution. Around 200 cells were imaged at 6–8 random areas of the µ-slide using wide field microscope Axio Observer Z1apotome (Zeiss, Jena, Germany) controlled by Zen software (Zeiss, Jena, Germany).

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RNA FISH Protocol for uc.333 and U6

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The probes to uc.333 RNA and U6 were designed following the Stellaris RNA fluorescence in situ hybridization probe designer (Biosearch Technologies). Cell fixation, permeabilization, and hybridization to probes were performed following protocols for adherent cells. Images were acquired with a Zeiss Axio Observer Z1 Apotome fluorescence microscope.

Zhang Y., Sun J., Yao H., Lin Y., Wei J., Hu G., Guo J, & Li J. (2020). Ultraconserved element uc.333 increases insulin sensitivity by binding to miR-223. Aging (Albany NY), 12(8), 6667-6679.

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Erythroid Cell A Antigen Expression Analysis

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Expression of A antigen was analysed by direct fluorescence staining with 50 μg/ml AF488‐conjugated helix pomatia agglutinin (HPA) lectin (ThermoFisher) in living cultured erythroid cells. As positive and negative controls, RBCs from individuals of A and O blood group types were fixed with 4% paraformaldehyde previous direct staining. Nuclei were stained with DAPI. Images were taken using Zeiss Axio Observer Z1 – Apotome inverted fluorescent microscope and analysed using the Image J software.¹⁹

Petazzi P., Miquel‐Serra L., Huertas S., González C., Boto N., Muñiz‐Diaz E., Menéndez P., Sevilla A, & Nogués N. (2022). ABO gene editing for the conversion of blood type A to universal type O in Rhnull donor‐derived human‐induced pluripotent stem cells. Clinical and Translational Medicine, 12(10), e1063.

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Visualizing uc.372 RNA and U6 via FISH

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The probes tiling the uc.372 RNA and U6 were designed following Stellaris^® RNA-fluorescence in situ hybridization probe designer (Biosearch Technologies). Cell fixation, permeabilization, and hybridization to probes were performed following protocols for adherent cells. Images were acquired on a Zeiss Axio Observer Z1 Apotome fluorescence microscope.

Guo J., Fang W., Sun L., Lu Y., Dou L., Huang X., Tang W., Yu L, & Li J. (2018). Ultraconserved element uc.372 drives hepatic lipid accumulation by suppressing miR-195/miR4668 maturation. Nature Communications, 9, 612.

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Live/Dead Cell Viability Assay

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Control and treated IGROV or IGROV-CP20 cells were gently washed with PBS. The live/dead reagents (Invitrogen) were dissolved in sterile PBS. Live/dead solution was added to the cells for 30 min to generate a green fluorescent signal in live cells and a red signal in dead cells. The labeled cells were viewed under the ZEISS Axio Observer.Z1 APOTOME fluorescence microscope using FITC and RFP filters to count live/dead cells in the treated and control specimens. Quantification was done in 10 fields for each condition and the proportion of live cells was calculated as % of total cells in the specimen.

Mariniello M., Petruzzelli R., Wanderlingh L.G., La Montagna R., Carissimo A., Pane F., Amoresano A., Ilyechova E.Y., Galagudza M.M., Catalano F., Crispino R., Puchkova L.V., Medina D.L, & Polishchuk R.S. (2020). Synthetic Lethality Screening Identifies FDA-Approved Drugs that Overcome ATP7B-Mediated Tolerance of Tumor Cells to Cisplatin. Cancers, 12(3), 608.

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Mitochondrial Dynamics in Benzo[a]pyrene-Treated Cells

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HCoEpC were grown on cover glasses, treated with Benzo[a]pyrene (5 µM) and/or Hydroxytyrosol (1 µM) for 6 days, and mitochondrial labelling was performed by adding MitoTracker^TM Green-FM (Invitrogen, Cat# M7514, MA, USA) fluorescent dye (200 nM) to cell culture medium for the last 30 min at 37 °C. Then, cells were washed in PBS and cover glasses mounted with PBS:Glycerol (1:1) on microscope slides. Cells were analyzed with Apotome Axio Observer Z1 inverted microscope (Zeiss, Munich, Germany) equipped with an AxioCam MRM Rev.3 (Germany) at 40× magnification.

Santarelli R., Pompili C., Gilardini Montani M.S., Evangelista L., Gonnella R, & Cirone M. (2022). 3,4-Dihydroxyphenylethanol (DPE or Hydroxytyrosol) Counteracts ERK1/2 and mTOR Activation, Pro-Inflammatory Cytokine Release, Autophagy and Mitophagy Reduction Mediated by Benzo[a]pyrene in Primary Human Colonic Epithelial Cells. Pharmaceutics, 14(3), 663.

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Detailed Microscopy Acquisition Settings

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All images were acquired using the Axio Scan. Z1 (Zeiss). The acquisition settings are summarized in Table 2. However, other acquisition systems have been tested (Additional file 1: Table S1), such as an Apotome widefield (Apotome Axio Observer Z1, Zeiss), a confocal microscope (LSM 700 AxioObserver, Zeiss), and a spinning disk confocal (Cell Voyager CV1000, Yokogawa, Japan).Table 2

Acquisition settings of tested images for the developed analysis feature

Acquisition		Analysis
Acquisition		centro-nucleated fibers	Vessels	Satellite cells	Fiber type
Image dimensions	Z-Stack	No	No	No	No
	Channels	2	3	3	4
	Scaling xy (per pixel)	0.325 μm × 0.325 μm	0.325 μm × 0.325 μm	0.38 μm × 0.38 μm	0.325 μm × 0.325 μm
Acquisition information	Microscope	AxioScan.Z1	AxioScan.Z1	LSM 700	AxioScan.Z1
	Objective	20×	20×	40×	20×
Channel 1	Reflector	DAPI	DAPI	DAPI	DAPI
	Excitation wavelength	353	353	404	353
	Emission wavelength	465	465	444	465
Channel2	Reflector	FITC	Cy3	Cy3	Cy3
	Excitation wavelength	495	548	561	548
	Emission wavelength	519	561	575	561
Channel 3	Reflector		FITC	FITC	FITC
	Excitation wavelength		495	488	495
	Emission wavelength		519	517	519
Channel 4	Reflector				Cy5
	Excitation wavelength				650
	Emission wavelength				673

Mayeuf-Louchart A., Hardy D., Thorel Q., Roux P., Gueniot L., Briand D., Mazeraud A., Bouglé A., Shorte S.L., Staels B., Chrétien F., Duez H, & Danckaert A. (2018). MuscleJ: a high-content analysis method to study skeletal muscle with a new Fiji tool. Skeletal Muscle, 8, 25.

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Immunofluorescence Staining of iPSCs and Neurons

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iPSCs, NPCs, and 2D neurons were fixed with 4% paraformaldehyde for 15 min. After three washes with PBS, cells were permeabilized with 0.25% Triton X-100 for 15 min, blocked with 3% bovine serum albumin (BSA) and incubated overnight at 4 °C with primary antibodies diluted in 3% BSA. The following day, cells were washed and incubated with the secondary antibodies for 1 h. Antibodies used in this study can be found in Table S9. For nuclei staining, DAPI solution (1 μg/mL) was used. The slides were mounted using ProLong Gold antifade reagent and analyzed under a fluorescence microscope (Z1 Axio Observer Apotome, Zeiss).

Negraes P.D., Trujillo C.A., Yu N.K., Wu W., Yao H., Liang N., Lautz J.D., Kwok E., McClatchy D., Diedrich J., de Bartolome S.M., Truong J., Szeto R., Tran T., Herai R.H., Smith S.E., Haddad G.G., Yates JR 3.r.d, & Muotri A.R. (2021). Altered network and rescue of human neurons derived from individuals with early-onset genetic epilepsy. Molecular Psychiatry, 26(11), 7047-7068.

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