A. Use frosted-end slides that have been rinsed with dH2O and
stored at 4 C. It may be helpful to warm the slides to room temperature before
B. Centrifuge the cultures for 6 minutes at 150 x
C. Remove all but 0.2 - 0.4 ml of supernatant. Resuspend the cell
D. Add fixative to dilute the cell suspension, 0.2 - 1 ml may be
required depending on the quantity of cells.
E. Using a pasteur pipet,
place 2 or 3 drops of cell suspension near the frosted end of the slide. Spread
cells by tilting the slide and blowing gently. Wipe dry the back of the slide
and place it on the slide warmer at 38 C for 1- 2 minutes until the slide is
dry. It may be necessary to adjust the temperature of the wet slides and time on
slide warmer to maximize cell spreading.
F. Label slides with patient
number, culture identification letter, and slide number.
G. Note the
number of slides made from each tube on the tape label of the tube.
possible, allow slides to age overnight at room temperature (for rush patients,
that may not be possible.) Slides are now ready for G, Q, C, or NOR
I. After making slides, add a pipetful of fixative to each tube,
cap tightly and store at 4 C for short term or -20 C for long term.