CaMKII isoforms and phosphoCaMKII (pCaMKII) were detected by indirect immunofluorescence as described in12 (link). For a list of retrieval methods, antibodies and dilutions please refer to Supplementary Table 2 . All images were acquired using identical settings on an Olympus BX61 microscope and consistently modified for best rendering using Adobe Photoshop. The fluorescent and dapi images of the same field were superimposed, or the fluorescent and Nomarski images of the same field were inverted and superimposed (subtraction).
Bx61 microscope
Manufactured by Adobe
Sourced in United States
The BX61 microscope is a high-performance optical microscope designed for a variety of laboratory applications. It features a stable and ergonomic design, providing a reliable platform for precise observation and analysis. The BX61 microscope is equipped with advanced optics, including objectives and eyepieces, to deliver clear and detailed images. It is suitable for a range of magnification levels to accommodate different sample types and research needs.
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2 protocols using bx61 microscope
1
Quantifying CaMKII and pCaMKII Localization
2
In situ hybridization of insect pheromone-binding genes
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For slnlg4-yll and slnrxI, cDNA fragments of respectively 748 bp and 565 bp were amplified by PCR using specific primers (Table S1), purified (Nucleospin Extract II, Macherey-Nagel) and cloned into pCRII-TOPO plasmid (Invitrogen), used as template for in vitro transcription to generate DIG-labeled RNA sense and antisense probes. The Pheromone-Binding protein SlPBP1 (GenBank accession number EF396284), whose transcripts are highly expressed in male antennae, was used as positive control. Antennae from three-day-old male moths were embedded in Tissue Tek 186 medium TM compound (CellPath, Newtown Powys, UK). Cryosections (7 µm) were set in cell culture insert (Greiner Bio-one, Monroe, USA). Hybridization was conducted as described previously (Durand et al. 2010a ). Pictures were acquired (Olympus BX61 microscope, ImagePro software) and digitalized using Adobe Photoshop ® 7.0 (Adobe, USA).
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