The roots from the transgenic seedlings (low density) were excised as described for quantifying the morphological traits. For histochemical analysis of the GUS activity in CycB1;1:CDB-GUS, pPIN1:GUS, pPIN2:GUS, pPIN3:GUS, pPIN4:GUS, and pPIN7:GUS, the excised roots of the transgenic seedlings were incubated overnight at 37 °C in a GUS reaction buffer (1 mg mL−1 5-bromo-4-chloro-3-indolyl-β-D-glucuronic acid, 5 mM each of K3Fe[CN]6 and K4Fe[CN]3H2O in 100 mM sodium phosphate buffer [pH 0]) as described [17 (link)]. Stained roots (~10–12) for each of the treatments were cleared with 70% (v/v) ethanol, and GUS activity in the primary root tip was captured by using a differential interference contrast (DIC) microscopy (Axio Imager 2, Carl Zeiss, Jena, Germany). The green fluorescent protein (GFP) images of the primary root tip of transgenics DR5:GFP were captured using Axio Imager 2 (Carl Zeiss) and merged with DIC images by employing ZEN lite 2012 analysis software [www.zeiss.com/microscopy/int/products/microscope-software/zen-lite.html , accessed on 11 August 2021].
Axio imager 2
Manufactured by Zeiss
Sourced in Germany, United States, Japan, United Kingdom, Canada, Switzerland
The Axio Imager 2 is a high-performance optical microscope system designed for advanced imaging and analysis applications. It features a stable, ergonomic design and offers a range of modular components to accommodate diverse research and industrial needs.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (27 mentions)
Apotome, by Zeiss (14 mentions)
Lsm 710, by Zeiss (14 mentions)
Lsm 700, by Zeiss (12 mentions)
Zen software, by Zeiss (10 mentions)
Rhodamine phalloidin, by Thermo Fisher Scientific (9 mentions)
Axiocam mrm camera, by Zeiss (8 mentions)
Discovery v20, by Zeiss (8 mentions)
Zen 2, by Zeiss (6 mentions)
Apotome 2 module, by Zeiss (6 mentions)
Lsm 780, by Zeiss (6 mentions)
Lsm 880, by Zeiss (6 mentions)
Axiovision 4, by Zeiss (6 mentions)
Axiocam digital camera, by Zeiss (5 mentions)
Apotome 2, by Zeiss (5 mentions)
Lsm 800, by Zeiss (5 mentions)
Axiocam mrm, by Zeiss (5 mentions)
Axiocam, by Zeiss (5 mentions)
Prolong diamond antifade mountant, by Thermo Fisher Scientific (4 mentions)
Axiocam mrc, by Zeiss (4 mentions)
439 protocols using axio imager 2
1
Histochemical Analysis of GUS and GFP Expression in Transgenic Arabidopsis Roots
2
Sclerotinia sclerotiorum Infection Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A 5 mm mycelia plug of S. sclerotiorum was placed on a glass slide and cultured for 24 h to observe the formation and number of appressoria. Samples were examined and photographed under stereo microscopes (Stemi508, ZEISS, Oberkochen, Germany) and a light microscope (Axio Imager 2, ZEISS, Oberkochen, Germany). After 16 h of inoculation with S. sclerotiorum, onion epidermis was soaked in 0.5% trypan blue solution for 30 min and then decolorized using bleaching solution (ethanol:acetic acid:glycerol = 3:1:1). Samples were examined and photographed under the light microscope (Axio Imager 2, ZEISS).
S. sclerotiorum was inoculated on PDA medium containing 100 μg/mL bromophenol blue to detect whether it secreted oxalic acid.
S. sclerotiorum was inoculated on PDA medium containing 100 μg/mL bromophenol blue to detect whether it secreted oxalic acid.
3
Muscle Fiber Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TA samples were mounted in tragacanth (Sigma-Aldrich # G1128) on plastic blocks and frozen in liquid isopentane cooled in liquid nitrogen and stored at -80°C. Samples were cut into 10μm cross-sections using a cryostat at -20°C then mounted on lysine coated slides (Superfrost), as described in.69 (link),73 (link) Cross-sections were brought to room temperature, rehydrated with PBS (pH 7.2), then blocked with goat serum (10% in PBS). They were then incubated with primary polyclonal anti-laminin rabbit IgG antibody (Sigma-Aldrich # L9393, 1:500) for 1 h at room temperature. Sections were then washed three times in PBS before being incubated for 1 h at room temperature with an Alexa Fluor® 594 goat anti-rabbit IgG antibody (A-11037, 1:500). Sections were then washed three times in PBS and slides were cover slipped using Prolong™ Gold (P36930, Invitrogen) as a mounting medium. Slides were imaged with a Zeiss fluorescence microscope (Zeiss Axio Imager 2). Median distribution of minimum Feret diameters of at least 300 fibers per muscle sample were analyzed using ImageJ (NIH, Bethesda, MD).74 (link) The degree of myofiber atrophy was calculated as percent difference in mean minimum Feret diameters, relative to muscles of sham Atg7f/f mice. The means, fibers distributions, and average number of fibers analyzed per muscle per mouse are shown in Figure S6 .
4
Single Cell Enumeration by Epifluorescence
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single cells were counted after in situ hybridization by epifluorescence microscopy according to the method described by Goulder (1977) (link). Triplicate filters for all samples were prepared for each dual hybridization conditions to minimize count variability and were subsequently counted in 100 independent microscopic fields. Blank samples were processed by treating 8 mL of 0.2 μm filtered PBS solution. Aggregate cells were counted as described by Lösekann et al. (2007) (link). Observations, enumerations, and images were performed with a Zeiss Axio Imager 2 epifluorescence microscope (Carl Zeiss) equipped with the slider module ApoTome® (Zeiss) at 100X magnification, the illumination system HXP-X and Colibri light technology (Zeiss) with appropriate filter sets for Cy3, Alexa Fluor 488, GFP, and DAPI fluorescence using an AxioCam MRm camera (Zeiss). Epifluorescence acquisitions were analyzed using the ZEN Pro 2012 software (Zeiss).
5
Punch Biopsy Analysis of Graft Tissue Morphology
6
Quantitative Analysis of Neuronal Markers in Insular Cortex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescent images were acquired at 40× magnification using a fluorescent microscope (Zeiss Axio Imager 2, Carl Zeiss Microscopy), and image processing was performed in FIJI (NIH, version 1.0). For each aIC and pIC subdivision (agranular, dysgranular, and granular), a squared area (200 × 200 μm) was selected to cover layers II/III. The number of neurons showing expression of pCREB, c-Fos, GAD67, or NeuN was counted manually. For colocalization of c-Fos with pCREB and pCREB with GAD67 immunoreactivity, the number of colocalized nuclei was counted in each aIC subdivision, then averaged per subdivision for each rat. For quantitative analysis of GAD67-positve puncta, images were acquired at 63× magnification, and the number of GAD67-positive puncta per pCREB-positive nucleus was counted manually in layers II/III of each aIC subdivision and then averaged per subdivision for each image.
7
Immunostaining of DU-145 Xenograft Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed paraffin-embedded DU-145 xenograft tissue sections were deparaffinized, rehydrated and subject to heat-induced antigen retrieval using antigen retrieval buffer (pH 6.0; Dako, Carpinteria, CA) in a pressure cooker. Sections were blocked with 2.5% normal horse serum for 30 min and incubated with mixture of rabbit monoclonal anti-CD133 antibodies (clone D2V8Q, diluted 1:100; Cell signaling, Danvers, MA) and mouse monoclonal anti-TRA (clone TRA-1-60 (S), diluted 1:1000; Cell signaling) or mixture of rabbit monoclonal anti-CD44 antibodies (clone E7K2Y, diluted 1:1000; Cell signaling) and mouse monoclonal anti-TRA (clone TRA-1-60 (S), diluted 1:1000; Cell signaling) for 1 h at room temperature, followed by incubation with mixture of Alexa Fluor 488 goat anti-rabbit IgG (ThermoFisher Scientific, Waltham, MA) and Alexa Fluor 555 goat anti-rabbit IgG (ThermoFisher Scientific). After nucleus visualization with DAPI, sections were mounted with Prolong Diamond antifade (ThermoFisher Scientific) and imaged on a Zeiss Axio Imager 2 (Carl Zeiss Microscopy, Jena, Germany).
8
Immunofluorescence Analysis of HDAC6 and SMAD7
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serial 3-μm sections from SS and LS patients were immersed in 5% PBS Bovine Serum Albumine (BSA) for 1 h at room temperature in order to prevent non-specific bindings. Samples were then incubated overnight at 4°C with HDAC6 antimouse and SMAD7 antigoat antibodies used at 1:100 dilution. After three washes in PBS for 10 min, the sections were incubated for 30 min at room temperature with secondary fluorescent antibodies donkey antigoat IgGFITC and goat antimouse IgG-TR (Santa Cruz Biotechnology Inc.) used at dilution 1:200 and 1:100 respectively. Nuclei were treated with 4’,6-diamidino-2-phenylindole (DAPI) at dilution 1:10, for 5 min at room temperature. Negative controls were obtained by keeping off the primary antibodies. Finally, the samples were observed under the Zeiss Axio Imager 2 with DFC 250 video camera (Carl Zeiss Microscopy, LLC, Thornwood, NY, USA). Microphotographswere processed with Image J software and analyzed by two pathologists (AV, RS) in a blinded fashion.
9
Visualizing Drosophila Larval Tracheal Development
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos from the appropriate crosses were collected on grape plates for four hours and after hatching, larvae were raised on mounds of yeast paste on grape plates at 22oC. For crosses involving btl-Gal4, UAS-Actin-GFP/CyO, GFP larvae were selected using a Leica MX FluoIII fluorescence microscope. For imaging, larvae were immobilized either with ether vapor before mounting in 70% glycerol or by heating for a few seconds on a 70oC hot plate [26 (link)] after mounting. Microscopes used for imaging were as follows; bright field and fluorescence images in Fig 1 (A-D, F, G) and Fig 5 (G, H)—Zeiss Axioplan2; bright field images in Fig 5 (A-D)—Zeiss Axioimager 2; bright field images in Fig 1(E) and Fig 5 (E, F)—Zeiss Axioskop. For quantitation of tracheal defects (Fig 5 ), larvae were imaged from Day1 after hatching until the day before pupation or death, as determined form the studies in Figs 3 and 4 . At least 10 larvae were imaged for each genotype and fluid-filling of the tracheae was quantitated from stored image sets.
10
Histological Assessment of Liver and Adipose Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A portion of the liver and adipose tissue was fixed with a 10-fold volume of 10% formalin. Thereafter, the tissues were embedded in paraffin blocks and sectioned using a cryo-cut microtome (Leica CM1800, Wetzler, Germany) with a thickness of 3–4 μm. The sections were stained with hematoxylin and eosin (H&E) and captured using an optical microscope (ZEISS Axio Imager 2, Carl Zeiss, Oberkochen, Germany). The liver histological changes were assessed (by a blinded observer) in three different randomly selected 20X fields for each experimental rat. Histological scores were measured using Kleiner’s histological scoring system [55 (link)] by quantifying the degree of inflammatory cell infiltration, steatosis, and balloon cells. Adipocyte size (μm2) and the number of crown-like structures (CLS) were determined in three different randomly selected 20X fields for each experimental rat. Adipocyte size and CLS number were measured using an Image J (NIH, Bethesda, MD, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!