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Axio imager z2 apotome microscope

Manufactured by Zeiss
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The Axio Imager Z2 Apotome microscope is a high-performance optical microscope designed for advanced imaging applications. It features an Apotome optical sectioning system that enables optical sectioning of samples, allowing for the acquisition of high-quality, three-dimensional images. The microscope is equipped with a range of advanced optics and illumination options to support a variety of imaging techniques.

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Lab products found in correlation

Axiocam mrm camera, by Zeiss (4 mentions) A 21449, by Thermo Fisher Scientific (2 mentions) Ab5733, by Merck Group (2 mentions) A 21244, by Thermo Fisher Scientific (2 mentions) M4439, by Merck Group (1 mentions) A11001, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Serva Electrophoresis (1 mentions) H6908, by Merck Group (1 mentions) A10040, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions)

7 protocols using axio imager z2 apotome microscope

1

Visualizing MEF2C and UBE3A Localization

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Expression plasmids expressing human MEF2C and UBE3A and respective negative control plasmids were obtained from Sino Biologicals (MEF2C-HA: HG12320-CY, UBE3A-Myc:HG11130-CM, pCMV-Myc:CV014 and pCMV-HA:CV013) and used for transient transfection. Transfected HeLa cells were grown on poly-lysine coated coverslips, fixated with 4% paraformaldehyde in PBS for 10 minutes and stained with anti-Myc (M4439, Sigma-Aldrich) and anti-HA (H6908, Sigma-Aldrich) and with Alexa Fluor™ 488 goat anti–mouse and Alexa Fluor™ 488 donkey anti–rabbit antibodies (A11001 and A10040, Thermo Fisher). Nuclei were counterstained with DAPI (Serva). Images were taken with a Zeiss Axio Imager Z2 Apotome microscope with a 63x objective and analyzed in ImageJ.
Straub J., Gregor A., Sauerer T., Fliedner A., Distel L., Suchy C., Ekici A.B., Ferrazzi F, & Zweier C. (2020). Genetic interaction screen for severe neurodevelopmental disorders reveals a functional link between Ube3a and Mef2 in Drosophila melanogaster. Scientific Reports, 10, 1204.
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2

Gut Dissection and H2DCFDA Staining

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Guts were dissected at the indicated times on glass slides, incubated in H2DCFDA (10 μM) for 15 min, and live imaged with a Zeiss Axio Imager Z2 Apotome microscope.
Ramond E., Jamet A., Ding X., Euphrasie D., Bouvier C., Lallemant L., He X., Arbibe L., Coureuil M, & Charbit A. (2021). Reactive Oxygen Species-Dependent Innate Immune Mechanisms Control Methicillin-Resistant Staphylococcus aureus Virulence in the Drosophila Larval Model. mBio, 12(3), e00276-21.
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3

Neurite Analysis of SK-N-BE (2) Cells

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For neurite analysis SK-N-BE (2) cells were transfected with 1 µg of mCD8-GFP using jetPRIME (Polyplus-transfection) following manufacturer’s instructions on day eight of differentiation. 24 h post transfection medium was changed. Cells were subjected to immunofluorescence analysis at differentiation day 10 as described above using only DAPI as counterstaining.
Cells were analyzed with a Zeiss Axio Imager Z2 Apotome microscope. Primary and secondary neurite length was measured using the NeuronJ plugin88 (link) in Fiji89 (link) and respective number was counted. Three PHF6 KO and three control cell lines were evaluated, for each line at least 30 cells were measured. The experiment was performed in biological triplicates. Data was tested for normality using the Kolmogorov–Smirnov test. Testing for statistical significance was performed using a two-tailed t test.
Fliedner A., Gregor A., Ferrazzi F., Ekici A.B., Sticht H, & Zweier C. (2020). Loss of PHF6 leads to aberrant development of human neuron-like cells. Scientific Reports, 10, 19030.
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4

Tracing Tanycyte Remodeling after Gene Delivery

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Four weeks after LV or 3V TAT-Cre infusion in TdTomato mice, animals were anesthetized with ketamine (8mg/kg)+xylazine (3mg/kg) before perfusion with 0.9% NaCl and 4% paraformaldehyde. Brains were collected, cryoprotected in 20% sucrose solution overnight, embedded in TissuTek (Sakura) and frozen. 16μm-thick coronal sections were cut and processed for immunofluorescence using chicken anti-vimentin (1:2000; Millipore Cat#AB5733, RRID:AB_11212377) primary and AlexaFluor-647-conjugated anti-chicken secondary antibody (1/1000; ThermoFisher Scientific Cat#A-21449, RRID:AB_2535866). Images were acquired using an AxioImager Z2 Apotome microscope (AxioCam MRm camera, Zeiss) and Zen 3.1 (blue edition) software. Eight representative ME slides/animal were coded to conceal treatment groups and tanycytes (vimentin-positive cells) divided into three groups depending on their projections (ME-ARH, ventromedial hypothalamus, dorsomedial hypothalamus). DAPI+/Tomato+/vimentin+ cell numbers was reported relative to DAPI+/vimentin+ cells bordering the 3V.
FACS-sorted Tomato-positive cell numbers were compared between microdissected samples of the ME (3V) and 4V, 1 week after infusion.
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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5

Visualizing pSTAT3-Positive Cells in Mouse Brain

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Adult mice were sacrificed by decapitation at lights-on. 20μm-thick coronal sections from fresh-frozen brains were postfixed in 2% paraformaldehyde during 1h and processed for immunofluorescence as described (Bouret et al., 2012) using rabbit anti-pSTAT3 (Tyr705) (1:1000; Cell Signaling Technology Cat#9131, RRID:AB_331586) primary and AlexaFluor-647-conjugated goat anti-rabbit secondary antibodies (1/500; Molecular Probes Cat#A-21244, RRID:AB_141663). Images were acquired using an AxioImager.Z2 Apotome microscope (AxioCam MRm camera, Zeiss). Slides were then coded to conceal treatment groups, and pSTAT3-immunoreactive cells counted in eight sections/animal.
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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6

Tracing Tanycyte Remodeling after Gene Delivery

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Four weeks after LV or 3V TAT-Cre infusion in TdTomato mice, animals were anesthetized with ketamine (8mg/kg)+xylazine (3mg/kg) before perfusion with 0.9% NaCl and 4% paraformaldehyde. Brains were collected, cryoprotected in 20% sucrose solution overnight, embedded in TissuTek (Sakura) and frozen. 16μm-thick coronal sections were cut and processed for immunofluorescence using chicken anti-vimentin (1:2000; Millipore Cat#AB5733, RRID:AB_11212377) primary and AlexaFluor-647-conjugated anti-chicken secondary antibody (1/1000; ThermoFisher Scientific Cat#A-21449, RRID:AB_2535866). Images were acquired using an AxioImager Z2 Apotome microscope (AxioCam MRm camera, Zeiss) and Zen 3.1 (blue edition) software. Eight representative ME slides/animal were coded to conceal treatment groups and tanycytes (vimentin-positive cells) divided into three groups depending on their projections (ME-ARH, ventromedial hypothalamus, dorsomedial hypothalamus). DAPI+/Tomato+/vimentin+ cell numbers was reported relative to DAPI+/vimentin+ cells bordering the 3V.
FACS-sorted Tomato-positive cell numbers were compared between microdissected samples of the ME (3V) and 4V, 1 week after infusion.
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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7

Visualizing pSTAT3-Positive Cells in Mouse Brain

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Adult mice were sacrificed by decapitation at lights-on. 20μm-thick coronal sections from fresh-frozen brains were postfixed in 2% paraformaldehyde during 1h and processed for immunofluorescence as described (Bouret et al., 2012) using rabbit anti-pSTAT3 (Tyr705) (1:1000; Cell Signaling Technology Cat#9131, RRID:AB_331586) primary and AlexaFluor-647-conjugated goat anti-rabbit secondary antibodies (1/500; Molecular Probes Cat#A-21244, RRID:AB_141663). Images were acquired using an AxioImager.Z2 Apotome microscope (AxioCam MRm camera, Zeiss). Slides were then coded to conceal treatment groups, and pSTAT3-immunoreactive cells counted in eight sections/animal.
Duquenne M., Folgueira C., Bourouh C., Millet M., Silva A., Clasadonte J., Imbernon M., Fernandois D., Martinez-Corral I., Kusumakshi S., Caron E., Rasika S., Deliglia E., Jouy N., Oishi A., Mazzone M., Trinquet E., Tavernier J., Kim Y.B., Ory S., Jockers R., Schwaninger M., Boehm U., Nogueiras R., Annicotte J.S., Gasman S., Dam J, & Prévot V. (2021). Leptin brain entry via a tanycytic LepR:EGFR shuttle controls lipid metabolism and pancreas function. Nature metabolism, 3(8), 1071-1090.
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