Immunohistochemistry [IHC] was performed on 2-µm sections of formalin-fixed, paraffin-embedded tissue [FFPE] according to a standard protocol using the ABC-AP Kit, Alkaline Phosphatase [Universal] [VECTASTAIN®, identifier: AK-5200, Vector labs, USA]. Briefly, after retrieval, sections were exposed to the primary antibody diluted in Dako REAL diluent [S202230-2, Agilent Technologies, Waldbronn, Germany] for 15 min at room temerature [RT], followed by incubation with the biotinylated secondary antibody [30 min, RT] and an incubation with the Vectastain ABC-AP reagent. After a TBS wash, a 5-min exposure to the Vector Blue alkaline phosphatase substrate was done and followed by Tris-buffered saline [TBS] wash. A second retrieval was performed, followed by another primary antibody incubation including all steps described above. The sections were exposed to the ImPACT Vector Red alkaline phosphatase substrate solution for 1-5 min, washed with deionized water and finally haematoxylin-counterstained. Dilutions and antigen-retrieval strategies for antibodies are listed in Supplementary Table S2 . Imaging was done using an Axio observer Z1, which was connected to an Axiocam 506 colour [both Carl Zeiss, Jena, Germany].
Axiocam 506 colour
Manufactured by Zeiss
Sourced in Germany
The Axiocam 506 colour is a high-resolution digital camera designed for microscopy applications. It features a 6.45 megapixel CMOS sensor that can capture detailed images with a resolution of 2752 x 2208 pixels. The camera supports a wide range of bit depths, including 8-bit, 10-bit, and 12-bit, allowing for accurate color reproduction and image quality. The Axiocam 506 colour is compatible with a variety of microscope models and can be integrated into various imaging software platforms.
Automatically generated - may contain errors
Lab products found in correlation
Axio observer z1, by Zeiss (1 mentions)
Dako real diluent, by Agilent Technologies (1 mentions)
Axioskop 2 plus, by Zeiss (1 mentions)
Imager a2 microscope, by Zeiss (1 mentions)
Axio lab a1, by Zeiss (1 mentions)
Agarose, by Merck Group (1 mentions)
Planneofluar z 1x 0.25 fwd 56 mm objective, by Zeiss (1 mentions)
4 protocols using axiocam 506 colour
1
Multiplex Immunohistochemistry Protocol
2
Tissue Extraction and Fixation for CLSTN3 IHC
3
Fungal Morphological Identification Protocol
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After activating the pathogen, picked some hyphae with the inoculation needle from the culture and transferred them on a slide aseptically for morphological identification. Mycelial samples with conidiophores and conidia were observed under a Zeiss Axio Lab A1 light microscope (Carl Zeiss, Germany) and microscopic observations made with objectives of 10x, 20x, 40x and 100x oil immersion. All measurements and photographs were performed using a Zeiss Imager A2 microscope with an Axiocam 506 colour camera and integrated software. Microscopically, the characteristics of 30 conidia and conidiophores from the isolates were observed. Morphological identification was performed using the Gams and Hoozemans (1970) and Seifert and Gams (2011) (link) methods.
4
ABA-Induced Root Growth Inhibition Assay
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Seedlings were grown for six days on plates containing growth medium with the same composition as the liquid medium but containing in addition 0.8% Agarose (Sigma). Seedlings were transferred from regular plates to plates either 10 µM ABA or no ABA (GM). Positions of root tips were marked after transfer and then roots were allowed to grow for 24 hours. After 24 hours images were taken using a Zeis Axio Zoom.V16 equipped with a PlanNeoFluar Z 1x/0.25 FWD 56mm objective lens, a 25x/10foc eyepiece and a Zeiss Axiocam506 colour.
Root lengths were determined using the freehand drawing tool in Fiji Version 2.1.0. Three independent experiments were performed and for each treatment / genotype combination >15 seedling roots were measured per experiment. For calculation of relative root length, the following formula was applied:
Root lengths were determined using the freehand drawing tool in Fiji Version 2.1.0. Three independent experiments were performed and for each treatment / genotype combination >15 seedling roots were measured per experiment. For calculation of relative root length, the following formula was applied:
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