Briefly, cells were fixed for 30 min at room temperature (RT) with 2% paraformaldehyde (PFA) and 0.1% glutaraldehyde. After washing and blocking with 10% normal goat serum for 30 min at RT, samples were incubated overnight with the primary antibody (1:100) at 4 °C. After washing, cells were incubated with secondary antibody (1:300) and DAPI (1:50,000) for 1 h at RT. The mounting medium hardened overnight at 4 °C and the cells were imaged with a Leica DMI 6000B-CS/TCS SP8 laser confocal microscope (objective: HC PL APO CS2 63x/1.40 oil, Leica, Wetzlar, Germany). Image stacks with a size of 0.3 µm were analyzed with ImageJ software (version 1.51p 22, National Institute of Health), as previously described [11 (link)].
Pl apo cs2 63x 1.40 oil
Manufactured by Leica
Sourced in Germany
The PL APO CS2 63x/1.40 oil is a high-performance objective lens designed for microscopy applications. It features a numerical aperture of 1.40 and a magnification of 63x, making it suitable for a wide range of imaging tasks. The lens is optimized for use with oil immersion to achieve maximum resolution and light-gathering capabilities.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pl apo cs2 63x 1.40 oil
1
Immunofluorescence Imaging of Cells
2
Visualizing Lysosomal Dynamics in C. elegans
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For confocal microscopy vha-13, vha-5 and hlh-30 endogenously tagged with mNeonGreen were synchronized via egg and maintained at 25°C to induce mild stress.
hlh-30 worms were imaged as day 4 adults, while vha-13 and vha-5 worms as L4s, because of high mNeonGreen background expression at day 4 adulthood. Worms were anaesthetized with 40 μM sodium azide, mounted on slide with 2% agar pad and imaged with a Leica TCS SP8 microscope equipped with HC PL APO CS2 63X/1.40
Oil and white light laser. Images were analysed using photoshop or Fiji software. To quantitate acidic lysosomes, worms grown at 25°C were incubated 48 hours prior to imaging with 2 !M LysoTracker Red DND-99 (Life Technologies). Worms were imaged as day 4 adults as described above. Number of puncta in a predefined squared area in the intestine (approximately spanning the second to fourth gut cell) were counted.
hlh-30 worms were imaged as day 4 adults, while vha-13 and vha-5 worms as L4s, because of high mNeonGreen background expression at day 4 adulthood. Worms were anaesthetized with 40 μM sodium azide, mounted on slide with 2% agar pad and imaged with a Leica TCS SP8 microscope equipped with HC PL APO CS2 63X/1.40
Oil and white light laser. Images were analysed using photoshop or Fiji software. To quantitate acidic lysosomes, worms grown at 25°C were incubated 48 hours prior to imaging with 2 !M LysoTracker Red DND-99 (Life Technologies). Worms were imaged as day 4 adults as described above. Number of puncta in a predefined squared area in the intestine (approximately spanning the second to fourth gut cell) were counted.
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