Transformed roots expressing GFP were further identified by fluorescence detection using an MZ10F dissecting microscope from Leica (Solms, Germany) equipped with a mercury UV-lamp and an eGFP band-pass or a DsRed2 long-pass filter. Confocal Laser Scanning Microscopy (CLSM) was carried out on an Axiovert Observer microscope with an LSM800 confocal module from Zeiss (Jenna, Germany), using a 488 nm diode laser line for excitation and collecting an emission wavelength range of 515 ± 15 nm, with the pinhole set to 1.0 airy unit and detector settings set to collect the full dynamic range of emitted fluorescence.
Mz10f dissecting microscope
Manufactured by Leica
Sourced in Germany
The Leica MZ10F is a dissecting microscope designed for detailed observation and analysis. It features a high-performance optical system with up to 80x magnification and a wide field of view. The microscope is equipped with LED illumination for bright, uniform illumination of the specimen.
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3 protocols using mz10f dissecting microscope
1
GFP and DsRed2 Fluorescence Imaging
2
Cloning and Characterization of MtNAS2 Promoter
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MtNAS2 promoter region was obtained by amplifying the 1,940 bp upstream of the start codon using the primers indicated in Supplementary Table 2
3
Plant total RNA extraction and GUS assay
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Total RNA was prepared from plant tissues using TRI Reagent according to the manufacturer's instructions (Invitrogen). Reverse transcription was performed using standard procedures with Improm-II reverse transcriptase (Promega, www.promega.com) . PCR amplifications from the cDNA template were performed using gene-specific primers (see Table S1). UBQ10 was used as the reference gene to normalize the data.
Histochemical GUS activity assay GUS activity was assayed by incubating plant tissues in GUS staining buffer (Bu et al. 2014 ) for 3-6 hours at 37 °C. Plant material was cleared in 70% ethanol, and observed directly under a Leica MZ10F dissecting microscope (www.leicamicrosystems.com).
Histochemical GUS activity assay GUS activity was assayed by incubating plant tissues in GUS staining buffer (Bu et al. 2014 ) for 3-6 hours at 37 °C. Plant material was cleared in 70% ethanol, and observed directly under a Leica MZ10F dissecting microscope (www.leicamicrosystems.com).
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