Cells were grown to mid-logarithmic phase and fixed with 4% formaldehyde in 0.1 M potassium phosphate buffer. Cells were then converted to spheroplasts using 0.1 M potassium phosphate buffer containing 1.2 M sorbitol and 500 μg of zymolyase. Spheroplasts were washed in 2× SSC buffer and incubated overnight at 37 °C with Cy3-labeled oligonucleotide probe (5′-Cy3-ATG CTC TTG CCA AAA CAA AAA AAT CCA TTT TCA AAA TTA TTA AAT TTC TT-3′) that is complementary to the 5′ portion of ITS1 (Faza et al., 2012). DNA was stained with 0.5 μg/ml DAPI. Cells were visualized using THUNDER Imager 3D Assay (Leica, Germany) equipped with a HCX PL-APO Fluotar 100×/1.44 NA oil immersion objective (Leica, Germany). Images were acquired with a fitted digital sCMOS camera and LAS X premium software (Leica, Germany).
Hcx pl apo fluotar 100 1.44 na oil immersion objective
Manufactured by Leica
Sourced in Germany
The HCX PL-APO Fluotar 100×/1.44 NA oil immersion objective is a high-performance microscope objective designed for advanced imaging applications. It features a 100x magnification and a numerical aperture of 1.44, providing exceptional optical resolution and light-gathering capability. This objective is optimized for use with oil immersion techniques to achieve the highest possible image quality and contrast.
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2 protocols using hcx pl apo fluotar 100 1.44 na oil immersion objective
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Fluorescent In Situ Hybridization of Yeast Cells
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Fluorescent In Situ Hybridization of Yeast Cells
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Cells were grown to mid-logarithmic phase and fixed with 4% formaldehyde in 0.1 M potassium phosphate buffer. Cells were then converted to spheroplasts using 0.1 M potassium phosphate buffer containing 1.2 M sorbitol and 500 μg of zymolyase. Spheroplasts were washed in 2× SSC buffer and incubated overnight at 37 °C with Cy3-labeled oligonucleotide probe (5′-Cy3-ATG CTC TTG CCA AAA CAA AAA AAT CCA TTT TCA AAA TTA TTA AAT TTC TT-3′) that is complementary to the 5′ portion of ITS1 (Faza et al., 2012). DNA was stained with 0.5 μg/ml DAPI. Cells were visualized using THUNDER Imager 3D Assay (Leica, Germany) equipped with a HCX PL-APO Fluotar 100×/1.44 NA oil immersion objective (Leica, Germany). Images were acquired with a fitted digital sCMOS camera and LAS X premium software (Leica, Germany).
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