Pleural and pedal ganglia cDNA was prepared and semi-quantitative PCR was performed as described in Reissner et al. (2008) (link). To ensure that amplification was kept within linear range, the number of cycles was optimized for each primer pair. apCRNF sense or antisense riboprobes were generated by in vitro transcription from SP6 or T7 promoters. FISH of cultured SNs or MNs was performed as described in Lyles et al. (2006) (link). Images were acquired with an Olympus Fluoview laser-scanning microscope.
Fluoview laser scanning microscope
Manufactured by Olympus
The Fluoview laser-scanning microscope is a high-performance imaging system designed for advanced fluorescence microscopy. It utilizes laser excitation and scanning technology to capture detailed images of biological samples.
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2 protocols using fluoview laser scanning microscope
1
Semi-quantitative PCR and FISH Analysis
2
Immunofluorescence Imaging of RAW264.7 Cells
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RAW264.7 cells were seeded and allowed to adhere on coverslips in 24-well plates, followed by treatment with 20 μg/mL rRv0297PGRS with or without ethylene glycol-bis (β-aminoethyl ether)-N, N, N′, N′-tetra acetic acid (EGTA), and calcium chloride (CaCl2). After 2 hrs. of incubation, the cells were fixed with 3.7% paraformaldehyde for 15–20 min. followed by blocking for 10 min. with 0.1% TritonX-100/1% BSA/PBS. The cells were labeled with respective primary and Alexaflour-conjugated secondary antibodies [84 (link)]. Immunofluorescence imaging was done using an Olympus Fluoview Laser-scanning microscope.
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