Lance D. Miller

NIH-NCI-DCS-MSO

(adapted from J. DeRisi;

Pat Brown Lab, Stanford)

 

 

Coating Slides with Poly-L-Lysine

 

Important: When handling slides, always wear powder-free latex gloves that have been washed in soap and water.

 

  1. Place 120 Gold Seal microscope slides into stainless steel slide racks (30/rack x 4; Shandon).
  2. Prepare cleaning solution as follows:    CAUTION: EXOTHERMIC REACTION.

 

350 ml/dish

1400 ml (4 dishes)

100 ml ddH20

400 ml ddH20

50 g NaOH

200 g NaOH (dissolve in H20 before adding EtOH)

250 ml 95% EtO

1000 ml 95% EtOH

(DANGER: add VERY slowly!)

 

Stir and add double distilled water (~200 ml) until solution is clear.

  1. Add 350 ml cleaning solution to each of 4 glass dishes (PGC Sci.), submerge slide-filled racks, and shake for 2 hours.
  2. Discard cleaning solution, refill with ddH2O, and shake 2-5 minutes with vigorous plunging.
  3. Refill, shake, and plunge - four more times.
    Note: all residual NaOH must be removed from slides at this step!
  1. Prepare lysine solution as follows:

 

350 ml/dish

1400 ml (4 dishes)

35 ml poly-l-lysine (Sigma)

140 ml poly-l-lysine

35 ml PBS (TC grade)

140 ml PBS

280 ml ddH2O

1120 ml ddH2O

 

  1. Add 350 ml lysine solution to each dish, submerge slides, and shake for 1 hour.
  2. Discard lysine solution and rinse slides for 10-15 seconds in ddH2O.
  3. Quickly spin dry in centrifuge (Beckman GS 6KR) @ 550 rpm (~80 x g) for 3 minutes.
  4. OPTIONAL: If vacuum oven is accessible, place racks in oven @ ~40oC for 5 minutes with vacuum.
  5. Transfer slides immediately to a clean slide box (no cork liner!), seal box, and allow to age for at least 2 weeks at room temperature. (Slides must age at least 2 weeks before aquiring optimal hydrophobic character.)

 

See Post Processing of Glass Slides