Cells of C. albicans ATCC 10321 that were treated with essential oils were examined using the Leica TCS SP8 Confocal Laser Scanning Microscope (Leica Microsystems, Wetzlar, Germany) and the LAS X 2.0.2.15022 software (Leica Microsystems, Wetzlar, Germany) in the Laboratory of Microscopic Imaging and Specialized Biological Techniques, University of Łódź. The fluorescence excitation of oils was induced using UV (Leica Microsystems, Wetzlar, Germany) diode of 405 nm., while the detection was recorded by three separate detectors with 800 ± 5 gain and -0.5 offset: PMT 1 at a 430–480 nm (Blue Channel), PMT 3 at a 500–550 nm (Green Channel), and PMT 5 at a 600–650 nm (Red Channel). Moreover, a PMT Trans Channel was used to visualize the cells in transmitted light. Each sample was scanned in the ‘xyz’ axes to a depth of 30 µm (three-dimensional (3D) scan) while using HC PL APO CS2 63×/1.40 (Leica Microsystems, Wetzlar, Germany) Oil objective. Line Average of 4× was used to improve the quality of images. The images were visualised in 3D view using a surface and blend mode with an adjusted threshold for a cross-section of cells.
Hc pl apo cs2 63 1
Manufactured by Leica
Sourced in Germany
The HC PL APO CS2 63×/1.40 is a high-performance objective lens manufactured by Leica. It is designed for use in various microscopy applications. The lens features a numerical aperture of 1.40 and a magnification of 63×, providing high-resolution imaging capabilities.
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Lab products found in correlation
Hoechst 33342, by Thermo Fisher Scientific (2 mentions)
Las x 2, by Leica (1 mentions)
Tcs sp8 confocal laser scanning microscope, by Leica (1 mentions)
Evos floid cell imaging station, by Thermo Fisher Scientific (1 mentions)
Tcs sp8 x laser scanning microscope, by Leica (1 mentions)
Mitotracker deep red, by Thermo Fisher Scientific (1 mentions)
2 protocols using hc pl apo cs2 63 1
1
Visualizing Candida albicans Treated with Essential Oils
2
Immunocytochemistry Staining and Microscopy
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Immunocytochemistry staining was performed as described previously [16, (link)17] (link). MitoTracker Deep Red (100 nM; Thermo Fisher Scientific, Waltham, MA, USA) was added after hyperoxic treatment and cells were incubated for 20 min at 37°C. Primary and secondary antibodies used for immunocytochemistry in this study are listed in Table 2. Confocal imaging was performed using Leica TCS SP8 X laser scanning microscope (Leica Microsystems, Wetzlar, Germany), equipped with an HC PL APO CS2 63/1.40 oil immersion objective and a white light laser. The excitation wavelengths and emission detection ranges used were 405 nm and 412-460 nm for DAPI, 488 nm and 495-550 nm for Alexa488, 594 nm and 601-644 nm for Alexa594, and 644 nm and 651-700 nm for MitoTracker Deep Red, respectively. For cell morphology analysis on EVOS Floid Cell Imaging Station (Thermo Fisher Scientific, USA), live cells in the medium were stained with NAO (1,5 µM) and Hoechst 33342 (5 µg/ml; Thermo Fisher Scientific, USA) for 5 min, washed 2x with 1x PBS and maintained in 1x PBS+Mg/Ca.
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