Synchronized L1 stage pax-3(ga96) animals were obtained by allowing embryos to hatch on NGM plates with no food overnight. The following day the newly hatched L1 larvae were added to NGM plates with OP50 and allowed to feed until they reached the mid-L2 larval stage when Pn.p nuclei were observed and counted using Nomarski DIC optics on a Zeiss Axioplan2 microscope. To observe and count P cells and seam cells in pax-3(ga96); jcIs1 animals, L1 stage worms were obtained as described above and scored using fluorescent microscopy on a Zeiss Axioplan 2 microscope. To observe P cells in pax-3(RNAi) animals containing the ajm-1::gfp hypodermal junctional marker we placed starved L1 larvae on pax-3 RNAi-feeding plates and allowed animals to produce progeny. The progeny were allowed to develop to gravid adults and their embryos were isolated by standard bleach methods. These embryos were added to M9 buffer and allowed to hatch overnight. The next day, P cells were scored for the Gap and Dis phenotypes (described below). sIs12963 animals injected with pax-3 dsRNA were observed by picking progeny of injected animals and viewed using the Axioplan2 fluorescent microscope.
Axioplan 2 microscope
Manufactured by Zeiss
Sourced in Germany, United States, Canada, United Kingdom
The Axioplan 2 microscope is a high-performance optical instrument designed for advanced research and analysis applications. It features a sturdy, modular construction and a range of interchangeable optical components to accommodate diverse sample types and experimental requirements. The Axioplan 2 provides reliable and precise imaging capabilities, enabling users to observe and study specimens with clarity and accuracy.
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Lab products found in correlation
Axiovision software, by Zeiss (47 mentions)
Axiovision 4, by Zeiss (35 mentions)
Axiocam hrc camera, by Zeiss (20 mentions)
Axiocam mrm camera, by Zeiss (20 mentions)
Lsm 710 confocal microscope, by Zeiss (18 mentions)
Axiocam, by Zeiss (17 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (16 mentions)
Axiocam digital camera, by Zeiss (16 mentions)
Axiocam mrm, by Zeiss (14 mentions)
Apotome, by Zeiss (14 mentions)
Axiocam camera, by Zeiss (13 mentions)
Axiovision, by Zeiss (13 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (13 mentions)
Lsm 780 confocal microscope, by Zeiss (12 mentions)
Axiocam hrm camera, by Zeiss (12 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (12 mentions)
Lsm 510, by Zeiss (10 mentions)
Axiocam hrc, by Zeiss (9 mentions)
Axiocam ccd camera, by Zeiss (8 mentions)
Vectashield, by Vector Laboratories (8 mentions)
979 protocols using axioplan 2 microscope
1
Analyzing Pax-3 Mutant Phenotypes
2
Golgi Staining of Dendritic Morphology in hMMP-1 Transgenic Mice
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Golgi staining was performed on hMMP-1 Tg animals and WT littermate controls aged 3–4 months old using FD Rapid GolgiStain kit (FD NeuroTechnologies, Inc.; Columbia, Maryland) according to the manufacturer’s instructions. The brains were sliced on a vibratome (VT1000S; Leica) at 150 μm. Images of CA1 pyramidal neurons were taken in bright-field on an Axioplan2 Zeiss microscope at 63X or 100X. Images were coded, and dendritic spines counted in a blinded manner similar to previous protocols81 (link). Dendritic branching was assessed in a blinded manner according to previously described methods82 (link). Fully impregnated cerebral somatosensory cortical layer IV/V neurons were selected. Using a Zeiss Axioplan 2 microscope at 40X, a blinded investigator imaged multiple focal planes so that each basilar branch could be followed in its entirety. For each animal, a total of 9 neurons were evaluated to determine the number of primary, secondary, tertiary and quaternary branches.
3
Anatomical Characterization of Rice Leaves
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For leaf anatomical studies, free-hand vertical sections (vs) of the leaves of different types (haploid, dihaploid, or tetraploid) of anther culture-derived rice plants were stained with safranin and photographed using a Carl Zeiss Axioplan-2 microscope equipped with an automatic exposure system. Epidermal peels were obtained from fresh leaf materials following the standard method [19] for stomatal study. Briefly, 1-cm-long pieces of the collected leaves were scraped on their abaxial sides to remove most of the cells above the adaxial epidermis, and the isolated adaxial epidermis was then stained with 1% safranin for 30 s, washed thoroughly in distilled water, mounted with diluted glycerine and photographed using a Carl Zeiss Axioplan-2 microscope. All photographs were taken at a similar magnification (×1800).
4
Cell Size and Vascular Morphology Analysis
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Cell size and number calculations were performed by staining cells with Calcofluor White (1:20 in PBS) of various internodes and imaging on a Zeiss Axioplan 2 microscope. Sizes of cells were then measured using ImageJ and averaged using three biological replicates per target tissue. The length of each cell per micrometer of internode was used to estimate the number of cells in the total length of the internode.
For cell staining and morphology characterization, the center of mature internodes was harvested and stained with either 0.1% Toluidine Blue or Safranin O to visualize the cell wall. Images were obtained using a black and white microscope, which allowed for a greater contrast and better discrimination of cell wall and vascular bundle elements than with color images. For the abnormal vascular bundle phenotypes, total vascular bundle numbers were calculated in five images from mature internodes of both DYM and DDYM in three biological replicates and were marked as either ‘normal’ or ‘misshapen’. After summation, the numbers of misshapen vascular bundles were reported as a total percentage of all vascular bundles counted. Cellulose staining was achieved by staining cells with Calcofluor White (1:20 in PBS) for 2 min with a 5‐minute destaining step and imaged using a Zeiss Axioplan 2 microscope. Three replicates were used for this experiment.
For cell staining and morphology characterization, the center of mature internodes was harvested and stained with either 0.1% Toluidine Blue or Safranin O to visualize the cell wall. Images were obtained using a black and white microscope, which allowed for a greater contrast and better discrimination of cell wall and vascular bundle elements than with color images. For the abnormal vascular bundle phenotypes, total vascular bundle numbers were calculated in five images from mature internodes of both DYM and DDYM in three biological replicates and were marked as either ‘normal’ or ‘misshapen’. After summation, the numbers of misshapen vascular bundles were reported as a total percentage of all vascular bundles counted. Cellulose staining was achieved by staining cells with Calcofluor White (1:20 in PBS) for 2 min with a 5‐minute destaining step and imaged using a Zeiss Axioplan 2 microscope. Three replicates were used for this experiment.
5
Cytogenetic Analysis of Mouse Breast Tumors
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SKY karyotype and DNA fluorescence in situ hybridization (FISH) experiments were performed by the Mayo Clinic Cytogenetics Core using primary cultures derived from F1 mouse breast tumors and freshly prepared normal fibroblasts. Briefly, fresh tumors were mechanically dissociated and cultured in RPMI 10% FBS containing Penn-Strep for 24–48 h, any debris was removed, the media replaced, and adherent cells were cultured for 1–3 weeks until sufficiently confluent. Cells were stored in liquid nitrogen until they could be processed for SKY. Reagents for SKY were from Applied Spectral Imaging. SKY images were captured using an Axioplan 2 microscope (Zeiss) and GenASis software version 7.27 (Applied Spectral Imaging). FISH images were captured using an Axioplan 2 microscope (Zeiss) and CytoVision Imaging software version 7.4 (Leica). The mouse centromeric probe BAC RP23-209m4 located at 7qA2 was provided by the BACPAC Resource Center (https://bacpacresources.org/ ) at Children’s Hospital Oakland Research Institute (Oakland, CA) and used to test the hypothesis that the transgene integration site was adjacent. The plasmid pSV2neuT plasmid was obtained from Addgene (http://www.addgene.org/ ) and was a gift from Bob Weinberg34 (link),35 (link).
6
TUNEL Staining for Apoptosis Assessment
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Liver tissue was frozen in optimum cutting temperature compound (Sakura Finetek) on dry ice. Tissue was finely cut using a cryostat by the Comparative Pathology Laboratory at Baylor College of Medicine. We then performed TUNEL staining in our lab with use of Takara’s In Situ Apoptosis Detection Kit (Otsu, Japan). Briefly, tissue was fixed in acetone for 30 min and washed. Tissue was permeablized with buffer contained in kit for 5 min and incubated with labeling mix (containing TdT enzyme and fluorescein-dUTP) for 90 min at 37°C. Tissue was then washed and mounted with a medium containing DAPI. Fluorescent images were captured using a Zeiss Axioplan 2 microscope at 400X using OpenLab 3.1.5 software. For primary hepatocytes, the cells were fixed in 4% PFA for 1 hr and permeabilized (0.1% Triton-X100 in 0.1% sodium citrate) for 5 min. The cells were washed and stained with TUNEL reaction mixture (2.5 mM cobalt chloride, 0.4 U/μl Tdt enzyme, and 2 μM fluorescein-dUTP) for 60 min at 37°C. The cells were washed and stained with DAPI and counted with a Zeiss Axioplan 2 microscope.
7
Azan-stained Microscopy Imaging Protocol
8
Stem Anatomy Analysis of Upright and Inclined Trees
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For both the upright and horizontally inclined trees, the stem at 10 cm above soil level was used to prepare fresh 70-µm-thick sections (vibrating blade microtome Leica VT1000S, Wetzlar, Germany) stained with Safranin : Alcian Blue (1:2) for 30 s, washed and mounted in 50% glycerol, and imaged with a Zeiss Axioplan2 microscope, AxioCam HRc camera, and AxioVision V 4.8.2 software (Carl Zeiss Light Microscopy, Göttingen, Germany). For TW : OW ratio determination, stereomicroscope images were obtained with a Canon PowerShot G7 digital camera. The distance from the center of the pith to the cambium of both the upper (TW) and lower (OW) sides was used to calculate the TW/OW ratio. Images for vessel quantification were taken with 10× magnification on a Zeiss Axioplan2 microscope. Images just inwards of the cambium, taken for NW, OW, and TW, were analyzed using ImageJ (version 1.51j8 USA).
9
Evaluating Chromosome Fragmentation and Survival
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Evaluation of chromosome fragmentation/missegregation: cells were fixed with ethanol and stained with 4′,6‐diamidino‐2‐phenylindole (DAPI) essentially, as described (Alao et al., 2014 ). Images were obtained with a Zeiss AxioCam on a Zeiss Axioplan 2 microscope with a ×100 objective, using the appropriate filter (DAPI or DIC). For quantifications at least 200 cells/replicate was counted. Three independent experiments were quantified.
Survival assay with propidium iodide (PI): Live cells were, at the indicated time point, stained with 10 µg/mL PI and subjected tor analysis by microscopy. Images were obtained with a Zeiss AxioCam on a Zeiss Axioplan 2 microscope with a ×100 objective, using the appropriate filter (red fluorescence or DIC). For quantifications, at least 200 cells/replicate were counted. Three independent experiments were quantified.
Survival assay with propidium iodide (PI): Live cells were, at the indicated time point, stained with 10 µg/mL PI and subjected tor analysis by microscopy. Images were obtained with a Zeiss AxioCam on a Zeiss Axioplan 2 microscope with a ×100 objective, using the appropriate filter (red fluorescence or DIC). For quantifications, at least 200 cells/replicate were counted. Three independent experiments were quantified.
10
Azan-stained Microscopy Imaging Protocol
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Azan-stained sections were photographed using a Zeiss Axioplan 2 microscope (Zeiss, Göttingen, Germany). Immunolabelled cryosections and whole-mount preparations were analysed using a Zeiss 510 Meta laser-scanning microscope (Zeiss) and a Zeiss Axioplan 2 microscope (Zeiss), respectively.
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