Sections of 5 μm thickness were deparaffinized in xylene and rehydrated in graded alcohols. Immunohistochemistry was performed using the Mouse specific HRP/DAB (ABC) Detection IHC Kit (Abcam, Cambridge, UK) according to manufacturer's protocol. The antigen unmasking was achieved with MS-unmasker solution (DIAPATH, Martinengo, BG, Italy) in microwave. GLUD2 primary antibody (cat. number SAB1400112, Sigma Aldrich, St Louis, MO) was used at 1:150 dilution for 1 h at room temperature. Slides were developed with diaminobenzidine chromogen (DAB) (DAKO, Glostrup, DK) and counterstained with hematoxylin. Negative controls included the omission of the primary antibody. Slides were analyzed using the inverted microscope CARL ZEISS Axio Observer Z1FLMot, and images were taken with CARL ZEISS AXIOCAM Icc1 camera (Zeiss, Oberkochen, Germany).
Carl axio observer z1flmot
Manufactured by Zeiss
Sourced in Germany
The CARL ZEISS Axio Observer Z1FLMot is a motorized fluorescence microscope designed for advanced imaging applications. It features fully motorized components for automated control of the optical system.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using carl axio observer z1flmot
1
Immunohistochemical Analysis of GLUD2
2
Analyzing Cellular Morphology of shANK Cells
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To observe the morphology of both shANK and shCTRL cells, we spread a drop from each cell suspensions over the glass-slide. Then the slides were allowed to air dry for 30 min and fixed for 10 min with 4% formaldehyde. At the end the slides were stained with Papanicolaou stain and reviewed by a senior cytopathologist under the inverted microscope Carl Zeiss Axio Observer Z1FLMot (Carl Zeiss Microscopy GmbH, Oberkochen, GE) who evaluated the main characteristics of malignancy in shANK cells: nuclear pleomorphism, ratio, multinucleation, pleomorphism of nucleoli and presence of multiple nucleoli. Images were taken with Carl Zeiss AxioCam Icc1 camera (Carl Zeiss Microscopy GmbH). These data were then confirmed through flow cytometric analysis, where multinucleated cells were detected with a propidium iodide staining. Analysis of flow cytometry data was carried out with FCS-Express 4 software (De Novo Software).
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