102
ANITA RAUCH
6. Remove fixative and incubate another 5 min at room temperature.
7. Remove fIxative once again and incubate another 5 min at room temperature.
8. Remove fIxative and let the slide air dry.
9. If the slide is not further used within a few days, store it at -20°C.
Preparation of urinary cells
1. Transfer the not necessarily sterile, but fresh urine sample into a 50 ml
tube.
2. Centrifuge at 172 g for 8 min (if r=16 cm 1000 U/min).
3. Remove the supernatant leaving a few millimeters.
4. Take the sediment with the remaining supernatant in a pipette and
drop it onto a clean slide.
5. Let the slide air dry.
6. Put the slide into a Quadriperm tray.
7. Carefully overlay the slide with Otto' solution and incubate for 30-60
min at 37°e.
8. Remove Otto's solution carefully and overlay the slide with fIxative,
incubate at room temperature for 5 min.
9. Remove fIxative and incubate another 5 min at room temperature.
10. Remove fIxative once again and incubate another 5 min at room temperature.
11. Remove fIxative and let the slide air dry.
12. If the slide is not further used within a few days, store it at -20°e.
RNase/Pepsin pretreatment
RNase 1. Cover slide with prepared epithelial cells with 100 III RNase working
solution and a coverslip and incubate in a metal tray in a 37°C water
bath for 15 min.
ANITA RAUCH
6. Remove fixative and incubate another 5 min at room temperature.
7. Remove fIxative once again and incubate another 5 min at room temperature.
8. Remove fIxative and let the slide air dry.
9. If the slide is not further used within a few days, store it at -20°C.
Preparation of urinary cells
1. Transfer the not necessarily sterile, but fresh urine sample into a 50 ml
tube.
2. Centrifuge at 172 g for 8 min (if r=16 cm 1000 U/min).
3. Remove the supernatant leaving a few millimeters.
4. Take the sediment with the remaining supernatant in a pipette and
drop it onto a clean slide.
5. Let the slide air dry.
6. Put the slide into a Quadriperm tray.
7. Carefully overlay the slide with Otto' solution and incubate for 30-60
min at 37°e.
8. Remove Otto's solution carefully and overlay the slide with fIxative,
incubate at room temperature for 5 min.
9. Remove fIxative and incubate another 5 min at room temperature.
10. Remove fIxative once again and incubate another 5 min at room temperature.
11. Remove fIxative and let the slide air dry.
12. If the slide is not further used within a few days, store it at -20°e.
RNase/Pepsin pretreatment
RNase 1. Cover slide with prepared epithelial cells with 100 III RNase working
solution and a coverslip and incubate in a metal tray in a 37°C water
bath for 15 min.