A total of 4 × 104 cells were resuspended in 150 μL of phosphate-buffered saline (PBS) and centrifuged on glass slides using a Cytofuge® (Medite, Burgdorf, Germany) at 700× g for 10 min. Afterwards, May–Grünwald Giemsa staining was performed. In short, cells were stained in 0.25% (w/v) May–Grünwald solution (Roth, Karlsruhe, Germany) for 5 min, washed in distilled water and stained with 1:20 diluted GIEMSA solution (Roth) for 20 min. After a second wash step, the slides were dried overnight and embedded in Roti-Histokitt mounting solution (Roth, Karlsruhe, Germany). Pictures were taken using an Olympus IX71 with the CellSens Dimension imaging software (Olympus, Shinjuku, Japan).
Roti histokitt mounting solution
Manufactured by Carl Roth
Sourced in Germany
Roti-Histokitt is a mounting solution designed for use in histology and microscopy. It is a high-quality, non-aqueous medium that provides a clear, durable, and long-lasting mounting of histological samples on microscope slides.
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2 protocols using roti histokitt mounting solution
1
May-Grünwald Giemsa Staining Protocol
2
Macrophage Cytospin Staining Protocol
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3×104 macrophages were resuspended in 150 µL phosphate-buffered saline (PBS) and centrifuged on glass slides (Menzel) using a Cytofuge (Medite) for 10 min at 700×g. Slides were stained with a 0.25% (w/v) May-Grünwald solution (Roth, Karlsruhe, Germany) for 5 min, washed thoroughly, and stained for 20 min with 1:20 diluted GIEMSA solution (Roth). The slides were washed a second round and dried over night . until coverage in Roti-Histokitt mounting solution (Roth). All images were taken with an Olympus IX71 using the CellSens Dimension imaging software (Olympus, Hamburg, Germany).
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