Axioimager compound microscope

Manufactured by Zeiss

Sourced in United States

The AxioImager is a compound microscope manufactured by Zeiss. It is designed for high-quality imaging and analysis of biological and material samples. The AxioImager features advanced optics and illumination systems to provide clear and detailed observations.

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Lab products found in correlation

Axiovision 4, by Zeiss (3 mentions) Axiocam icc3 color camera, by Zeiss (3 mentions) Axiocam camera, by Zeiss (2 mentions) Topo ta cloning, by Thermo Fisher Scientific (1 mentions) Dig labeling mix, by Merck Group (1 mentions) T7 rna polymerase, by Promega (1 mentions) Sp6 rna polymerase, by Thermo Fisher Scientific (1 mentions) Dfc7000 t camera, by Leica (1 mentions) Stereomicroscope, by Leica (1 mentions) Sp8 upright confocal microscope, by Leica (1 mentions) Ab245880, by Abcam (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Plan neofluar 10x 0.3 na objective, by Zeiss (1 mentions) Discovery v20, by Zeiss (1 mentions) Fv1000, by Olympus (1 mentions) Axioplan 2 compound microscope, by Zeiss (1 mentions) Lsm 700 or 800 confocal microscope, by Zeiss (1 mentions)

Close spelling variants of this product

AxioImager microscope (407 citations) Compound microscope (36 citations) Axioimager Z1 compound microscope (7 citations) AxioImager A2 compound microscope (3 citations)

11 protocols using axioimager compound microscope

Whole Mount In Situ Hybridization in Zebrafish

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Whole mount in situ hybridization was performed as described before^{94 (link)}.
To synthesize bcl6b, cx30.3, esm1, otc and si:dkey-28n18.9 probes, partial cDNA was first amplified from the cDNA of 24 hpf embryos using reverse oligo containing T7 promoter sequence. Resulting PCR product was then used as a template for RNA synthesis. Following primers were used for amplification of partial cDNA (Table S8). Anti-sense RNA was synthesized using T7 RNA polymerase (Promega) and DIG-labeling mix (Sigma-Aldrich). To synthesize bambib, bmp16, notchl and wnt11r probes, partial cDNA sequence was amplified using gene specific primers (Table S8). cDNA for each gene was subcloned into PCRII-TOPO vector using TOPO T/A cloning (ThermoFisher). Plasmids were linearized and antisense RNA was synthesized using SP6 (bmp16, wnt11r) or T7 (bambib, notchl) RNA polymerase (Promega) and DIG-labeling mix (Sigma-Aldrich). WISH embryos were imaged with a Nikon Eclipse Ni-E or Zeiss Axio imager compound microscope. HCR v3.0 probes for cldn5b, dab2, lyve1b, glula and krt18 were obtained from Molecular Technologies, Inc., Los Angeles, CA, USA. HCR in situ was performed as described previously^{95 (link)}. Embryos were mounted in 0.6% low-melting agarose and imaged using Nikon Eclipse confocal microscope. At least 15 embryos processed for WISH and HCR were analyzed.

Gurung S., Restrepo N.K., Chestnut B., Klimkaite L, & Sumanas S. (2022). Single-cell transcriptomic analysis of vascular endothelial cells in zebrafish embryos. Scientific Reports, 12, 13065.

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In situ hybridization of etv2 in embryos

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DIG-UTP labeled riboprobes were synthesized using T3, T7, or SP6 RNA polymerases (ThermoFisher). In situ hybridization was performed as described [19 (link)] using a previously reported antisense etv2 probe [20 (link)]. Processed embryos were cleared in BBA (2:1 benzyl benzoate: benzyl alcohol) or RIMS (57% Histodenz, 0.2% Tween 20, 0.02% sodium azide in 0.02M sodium phosphate buffer, pH=7.4) for imaging of tumor vessels as needed. Z-stacks of images were captured using an AxioImager compound microscope (Carl Zeiss Inc., USA) equipped with a Plan-Neofluar 10X/0.3 NA microscope objective (Carl Zeiss Inc., USA) and an AxioCam ICC3 color camera (Carl Zeiss Inc., USA). Extended focus images were produced using AxioVision 4.6 software (Carl Zeiss Inc., USA). Image levels and color balance were adjusted using Adobe Photoshop CS5.

Baltrunaite K., Craig M.P., Desai S.P., Chaturvedi P., Pandey R.N., Hegde R.S, & Sumanas S. (2017). ETS transcription factors Etv2 and Fli1b are required for tumor angiogenesis. Angiogenesis, 20(3), 307-323.

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Multicolor Confocal Imaging of Limb Development

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Secondary antibodies were as follows: goat anti-chicken IgY (H+L) secondary antibody, Alexa Fluor 488 (Invitrogen, A11039, 1:500), goat anti-mouse IgG (H+L) cross-adsorbed ReadyProbes secondary antibody, Alexa Fluor 594 (Invitrogen, R37121, 1:500) and goat anti-mouse IgG (H+L) cross-adsorbed ReadyProbes secondary antibody, Alexa Fluor 488 (Invitrogen, R37120, 1:500).
A Leica SP8 upright confocal microscope with a 40×/1.3 HC PL Apo CS2 Oil objective was used for all confocal images except for Fig. S9B,C, which were taken with a Leica SP8 inverted confocal microscope with a 20×/0.75 HC PL Apo CS2 Multi. LAS X was used for setting tiled images and a 20% overlap between tiles was used. Limb whole-mount HCR images were taken via a Leica stereomicroscope equipped with a DFC7000T camera. Fiji was used for maximum projection of z-stacks and to adjust contrast to highlight biological relevance. If needed, images were cropped, flipped and/or rotated to highlight biological relevance.
Histological staining can be carried out on cryosectioned HCR samples. Briefly, samples were stained with Hematoxylin and Eosin according to manufacturer's protocol (Abcam, ab245880) then samples were stained for Alcian Blue (Sigma, B8438) according to manufacturer's protocol. Histology images were captured on a Zeiss AxioImager compound microscope.

Aztekin C., Hiscock T.W., Gurdon J., Jullien J., Marioni J, & Simons B.D. (2021). Secreted inhibitors drive the loss of regeneration competence in Xenopus limbs. Development (Cambridge, England), 148(11), dev199158.

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In situ Hybridization of Vascular Markers

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In situ hybridization was performed as previously described (Jowett, 1999 (link)). Anti-sense riboprobes for kdrl, fli1a, vegfc (Cermenati et al., 2013 ), kdr (Covassin et al., 2006 (link)), and flt4 labeled with DIG-UTP were synthesized using T7 RNA polymerase (Ambion/Promega) as previously described (Thompson et al., 1998 (link)). vegfaa probe was synthesized from PCR products derived from pCS2-vegfaa₁₂₁ plasmid (Liang et al., 2001 (link)) using the primers below designed with an integrated T7 promoter site. Processed embryos were dehydrated in 100% ethanol for storage at −20°C to improve contrast then rehydrated in PBS and mounted in 3% methylcellulose under cover slips for imaging. An AxioImager compound microscope with Plan-Neofluar 10X/0.3 NA objective and AxioCam ICC3 color camera (Carl Zeiss Inc., USA) controlled using AxioVision 4.6 software (Carl Zeiss Inc., USA) was used to capture Z-stack images and produce extended focus images. Image contrast and brightness was adjusted using Adobe Photoshop CS6. Primer sequences: vegfaa probe fw:

Koenig A.L., Baltrunaite K., Bower N.I., Rossi A., Stainier D.Y., Hogan B.M, & Sumanas S. (2016). Vegfa signaling promotes zebrafish intestinal vasculature development through endothelial cell migration from the posterior cardinal vein. Developmental biology, 411(1), 115-127.

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Measuring C. elegans Embryo and Larval Size

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Progeny were collected by hypochlorite treatment of worms after 96 hr of culture. Embryo length measurements are constant during development due to the rigid eggshell, so developmental stage is not a confounding factor. To measure embryo size, embryos were plated onto unseeded 10 cm plates with NGM agar. These were imaged with a Zeiss Discovery.V20 stereomicroscope with a 10x objective (KSC 190–975). The images were analyzed using FIJI and calibrated to a micrometer. Lengths of embryos were measured both manually and by automatically thresholding embryos and calculating the long axis from ellipse fitting. Area and width measurements were also generated by thresholding. Specifically, background was subtracted, images were thresholded, they were converted to binary, holes were filled, and then particles were analyzed. This analysis was done in batch and the results were manually curated to ensure only quality embryo images were used.
Size of L1 larvae and worms cultured for 48 or 96 hr (starting from L1 arrest) was measured using the Wormsizer software [64 (link)]. Worms were washed with S-basal and plated on unseeded 10 cm NGM plates. These were imaged with a Zeiss Discovery.V20 stereomicroscope and magnification was adjusted based on the stage of the worms. Representative images of adult worms were taken at 10x on a Zeiss AxioImager compound microscope.

Hibshman J.D., Hung A, & Baugh L.R. (2016). Maternal Diet and Insulin-Like Signaling Control Intergenerational Plasticity of Progeny Size and Starvation Resistance. PLoS Genetics, 12(10), e1006396.

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Serial Sectioning of Fish Tissues

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Mutant and pharmacologically treated fish, as well as their controls, were sacrificed according to the Belgian law on the protection of laboratory animals (KB d.d. 13 September 2004) by an overdose of the anaesthetic MS222, fixed in paraformaldehyde-glutaraldehyde, embedded in epon, serially sectioned into 1 μm cross sections, stained with toluidine blue and mounted in DePex, as described previously (Huysseune and Sire, 1992 (link)). Specimens used for ISH were likewise sacrificed and, following ISH, embedded in epon and serially sectioned into 4 μm cross sections, as described in Verstraeten et al. (2012 ). Serial sections are required for detecting young tooth germs prior to any matrix deposition, as well as to identify their developmental stage and the tissue layers showing expression. Inevitably this procedure reduces the number of specimens that can be analyzed within reasonable time limits.
All sections were examined using a Zeiss Axio Imager compound microscope. Photographs were made with an Axio MRC camera. A schematic representation of the dentition of each specimen was obtained based either on superimposition of drawings made by camera lucida, or on serial photographs.

Huysseune A., Soenens M, & Elderweirdt F. (2014). Wnt signaling during tooth replacement in zebrafish (Danio rerio): pitfalls and perspectives. Frontiers in Physiology, 5, 386.

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Whole Mount in situ Hybridization in Zebrafish

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Whole mount in situ hybridization (ISH) was performed as previously described (Jowett, 1999 (link)). DIG-labeled antisense RNA probes for etv2, lyve1b, and flt4 were synthesized as previously described (Hogan et al., 2009a (link); Sumanas et al., 2005 (link); Thompson et al., 1998 (link)). Prior to imaging, zebrafish embryos or larvae were dehydrated to 100% ethanol, slowly rehydrated, whole mounted in 3% methylcellulose or 0.6% low melting agarose and imaged with the AxioImager compound microscope (Carl Zeiss) equipped with a Plan-Neofluar 10×/0.3 N.A. microscope objective (Carl Zeiss) and an AxioCam Icc3 color camera (Carl Zeiss, Inc.). A series of z-slices were acquired using AxioVision 4.6 software (Carl Zeiss, Inc.) to produce extended focus projected images.

Davis J.A., Koenig A.L., Lubert A., Chestnut B., Liu F., Desai S.P., Winkler T., Pociute K., Choi K, & Sumanas S. (2018). ETS transcription factor Etsrp / Etv2 is required for lymphangiogenesis and directly regulates vegfr3 / flt4 expression. Developmental biology, 440(1), 40-52.

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Quantitative Imaging of Tracheal Epithelium

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z stacks of the full epithelial thickness were acquired at an optical resolution of 1,024 × 1,024 with an optical z slice every 1 μm. Clones were scored manually by looking through the entire z depth of the tracheal epithelium in FV viewer or LAS AF software to score the identity of all labeled cells. For tetO-H2B-GFP samples, z stacks were acquired at an optical resolution of 1,024 × 1,024, with a z slice every 0.38 μm. Fluorescence intensity was assessed in Fiji, using the Gurdon Institute Imaging Facility’s plugin, ObjectScan.
Cryosections for analysis of cellular composition/density were imaged on an Olympus FV1000, using a 100× oil objective (numerical aperture [NA] 1.4). The length of the basement membrane in each image was measured in Fiji. Density was calculated as the number of cells present per μm of basement membrane. Cryosections for BrdU analysis were imaged on a Zeiss AxioImager compound microscope, using a 20× air objective (NA 0.8) and counted in Fiji.

Watson J.K., Rulands S., Wilkinson A.C., Wuidart A., Ousset M., Van Keymeulen A., Göttgens B., Blanpain C., Simons B.D, & Rawlins E.L. (2015). Clonal Dynamics Reveal Two Distinct Populations of Basal Cells in Slow-Turnover Airway Epithelium. Cell Reports, 12(1), 90-101.

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Quantifying Embryonic VIT-2 Expression

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Embryos were prepared by standard hypochlorite treatment of worms raised in AL or DR liquid culture conditions for 96 hr and were mounted on 4% noble agar pads. Images were taken at 1000x magnification using an AxioImager compound microscope equipped with an AxioCam camera (Zeiss). VIT-2::GFP was quantified using the ‘Measure’ function in Image J. A threshold that included the embryos but not the surrounding agar was manually identified for each reporter strain and culture system (AL/DR liquid culture and RNAi on plates). That threshold was used to remove background for all embryos. Average pixel intensity for the area above background was reported.

Jordan J.M., Hibshman J.D., Webster A.K., Kaplan R.E., Leinroth A., Guzman R., Maxwell C.S., Chitrakar R., Bowman E.A., Fry A.L., Hubbard E.J, & Baugh L.R. (2019). Insulin/IGF signaling and vitellogenin provisioning mediate intergenerational adaptation to nutrient stress. Current biology : CB, 29(14), 2380-2388.e5.

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Worm Imaging at 400x Magnification

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Worms were imaged at 400x magnification, using an AxioCam camera equipped with Zen software, on an AxioImager compound microscope (Zeiss). Images were merged using Fiji. Images were cropped and features were added using Inkscape.

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