Cells were cultured at 25°C in standard rich media (YPD) and collected at OD 0.5–1, fixed in 4% formaldehyde/YPD for 2 hours, washed twice in PBS, and stained overnight with 250 nM phalloidin-Alexa568 (Invitrogen Ref. A12380; Carlsbad, CA, USA). Cells were imaged using a Leica TCS SP8 X White Light Laser confocal microscope (Leica, Wetzlar, Germany) equipped with an HC PL APO CS2 100×/1.4 NA oil objective and a hybrid detector. Z-stack images were collected every 0.3 μm with Las X 3.5.5.19976 software. Patches were counted manually from the maximum intensity z-stack projections.
Hc pl apo cs2 100 1.4 na oil objective
Manufactured by Leica
Sourced in Germany
The HC PL APO CS2 100×/1.4 NA oil objective is a high-performance objective lens designed for Leica microscopes. It offers a magnification of 100x and a numerical aperture of 1.4, which provides high-resolution imaging and excellent light-gathering capabilities. The objective is optimized for use with oil immersion, allowing for enhanced contrast and resolution when observing samples.
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2 protocols using hc pl apo cs2 100 1.4 na oil objective
1
Actin Cytoskeleton Imaging in Yeast
2
Imaging Actin Dynamics in Yeast
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Log phase cultures in YPD medium at 25°C were fixed with 4% formaldehyde for 2 h. For CK‐666‐sensitivity assays, cells were treated with the indicated concentration of CK‐666 (Sigma‐Aldrich SML0006) for 30 min before fixation. After fixation, cells were washed twice in PBS and stained overnight with 250‐nM Phalloidin‐Alexa568 (Invitrogen, ref. A12380) at 25°C. Samples were washed twice with PBS, resuspended in PBS‐70% glycerol and directly mounted for imaging. Cells were imaged using a Leica TCS SP8 X White Light Laser confocal microscope equipped with an HC PL APO CS2 100×/1.4NA Oil objective and a hybrid detector. Z‐stack images were collected every 0.3 µm with Las X 3.5.5.19976 software.
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