50,000 – 100,000 cells were harvested, washed once at 300 × g for 5 min, resuspended in 200 μLof FACS Buffer, and spun onto poly-L-lysine coated microscope slides with a Shandon 4 (Thermo Scientific) cytocentrifuge at 300 rpm for 4 min. When visibly dry slides were transferred into May-Grünwald solution (Sigma-Aldrich) for 5 min, rinsed 4 times for 30 s in water, and transferred to Giemsa solution (Sigma-Aldrich) for 15 min. Slides were washed as described above, dry mounted with coverslips, and examined. All images shown were taken using a Metafer slide scanning platform and software (Metasystems) at 63X magnification.
Shandon 4
Manufactured by Thermo Fisher Scientific
The Shandon 4 is a high-quality tissue processor designed for routine and special tissue processing. It features a compact, easy-to-use design with a capacity of up to 100 cassettes. The Shandon 4 provides reliable and consistent tissue processing to support laboratory workflows.
Automatically generated - may contain errors
Lab products found in correlation
May gr nwald solution, by Merck Group (3 mentions)
Metafer slide scanning platform and software, by MetaSystems (3 mentions)
Giemsa solution, by Merck Group (3 mentions)
Poly l lysine coated glass slide, by Merck Group (1 mentions)
Axiovision software, by Zeiss (1 mentions)
Hoechst 33342 b2261, by Merck Group (1 mentions)
Dako fluorescent mounting medium, by Agilent Technologies (1 mentions)
Bx53 fluorescence microscope, by Olympus (1 mentions)
5 protocols using shandon 4
1
Cell Preparation for Microscopy Analysis
2
Cell Preparation for Microscopy Analysis
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50,000 – 100,000 cells were harvested, washed once at 300 × g for 5 min, resuspended in 200 μLof FACS Buffer, and spun onto poly-L-lysine coated microscope slides with a Shandon 4 (Thermo Scientific) cytocentrifuge at 300 rpm for 4 min. When visibly dry slides were transferred into May-Grünwald solution (Sigma-Aldrich) for 5 min, rinsed 4 times for 30 s in water, and transferred to Giemsa solution (Sigma-Aldrich) for 15 min. Slides were washed as described above, dry mounted with coverslips, and examined. All images shown were taken using a Metafer slide scanning platform and software (Metasystems) at 63X magnification.
3
Cell Preparation and Staining Protocol
4
Cell Preparation for Microscopic Analysis
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Approximately 50,000–200,000 cells were harvested, washed once at 300 x g for 5 min, resuspended in 200 µL FACS buffer and spun onto poly-L-lysine coated glass slides (Sigma Aldrich) with a Shandon 4 (Thermo Fisher) cytocentrifuge at 300 rpm for 4 min. Visibly dry slides were stained with May-Grünwald solution for 5 min, rinsed four times for each 30 s in H2O, transferred to Giemsa solution for 15 min and washed as described above. Slides were dried overnight and mounted with coverslip. All images were taken with AxioVision software (Zeiss) at 100 x magnification.
5
Analyzing Nuclear Morphology of Cells
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The nuclear morphology of control and 5,6 α-EC or 5,6 β-EC-treated cells was analyzed by fluorescence microscopy after staining with 1 µg/mL of Hoechst 33,342 (B2261, Sigma-Aldrich, Saint Louis, MO) [19 (link)]. Almost 4 × 104 cells were applied to glass slides by 5 min cytocentrifugation with a cytospin Shandon 4 (Thermo Fisher Scientific, Waltham, MA). Slides were mounted with the Dako fluorescent mounting medium (S302380, Dako, Copenhagen, Denmark) and observed under an Olympus BX53 fluorescence microscope (Olympus Corporation, Tokyo, Japan).
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