Protoplasts were prepared according to Yoo et al (2007) (link). The youngest fully expanded leaves (3rd or 4th true leaf for Col-0 and 1st or 2nd true leaves for slo3) were used. Protoplasts were stained with 1/2,000 dilutions of SYBR Green I (TaKaRa, https://www.takarabio.com ) and 1 µM mitoTracker Orange CMTMRos (ThermoFisher Scientific). Protoplasts were observed using a Leica STELLARIS5 confocal microscope (Leica) with a HC PL APO CS2 20× DRY objective lens (N.A. = 0.75) and HyD detectors. The Leica STELLARIS5 microscope was controlled by LAS × software (Leica), SYBR Green I and mitoTracker Orange CMTMRos were excited with 488 and 561 nm tuned white light laser, respectively. The fluorescence emissions were detected through a band-pass filter at 503–570 nm with 6.25% gain and a filter at 583–640 nm with 1.25% gain for SYBR Green I and mitoTracker Orange CMTMRos, respectively. The obtained images were analyzed by Image J for mitochondria and nucleoids.
Hc pl apo cs2 20 dry objective lens
Manufactured by Leica
The HC PL APO CS2 20× DRY objective lens is a high-performance objective lens designed for use with Leica microscopes. It features a 20× magnification and a dry working distance, making it suitable for a variety of microscopy applications. The lens is part of the Leica HC PL APO series, which is known for its excellent optical performance and chromatic aberration correction.
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Lab products found in correlation
Stellaris 5 microscope, by Leica (2 mentions)
Stellaris 5 confocal microscope, by Leica (2 mentions)
Sybr green 1, by Takara Bio (1 mentions)
Mitotracker orange cmtmros, by Thermo Fisher Scientific (1 mentions)
Las software, by Leica (1 mentions)
Las x software, by Leica (1 mentions)
Mc170 hd, by Leica (1 mentions)
2 protocols using hc pl apo cs2 20 dry objective lens
1
Visualizing Mitochondria and Nucleoids in Protoplasts
2
Visualizing Sorghum and Onion Cell Structures
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Enlarged images of the secondary branches and spikelets of sorghum and Kasalath were captured under a stereomicroscope (M125; Leica, Wetzlar, Germany) using a CCD camera (MC170 HD; Leica). Onion epidermal cells expressing GFP and DAI-GFP under control of the 35S promoter and calcofluor white-stained dissected awns were observed using a Leica STELLARIS5 confocal microscope (Leica) with an HC PL APO CS2 20× DRY objective lens (numerical aperture = 0.75), HC PL APO CS2 63× OIL objective lens (numerical aperture = 1.40), and HyD detectors. The Leica STELLARIS5 microscope was controlled using the LAS X software (Leica); 4′,6-diamidino-2-phenylindole dihydrochloride/calcofluor white and GFP were excited using 405 nm diode laser and 495 nm tuned white light lasers, respectively.
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