Slide Preparation

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 making slides.

B. Centrifuge the cultures for 6 minutes at 150 x g.

C. Remove all but 0.2 - 0.4 ml of supernatant. Resuspend the cell pellet.

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.

H. If 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 banding.

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.