Coating Beads on Coverslip

Procedure

Step 1. Prepare clean coverslip

  1. Clean the coverslip with HCl or NaOH.

  2. Wash several times with ddH2O.

  3. Store the coverslip in EtOH.

  4. Wash with ddH2O before use.

Step 2. Coating coverslip with Poly-L-Lysine

  1. Dilute Poly-L-lysine Solution 1:10 with deionized water. Results in 10% Poly-L-lysine.

  2. Embed coverslip in 10% Poly-L-lysine (~25uL for 5mm in diameter coverslip) for 20 mins.

  3. Wash several times with ddH2O.

Step 3. Coating beads

For dense bead sample

  1. Dilute the stock bead solution 1:1000 with deionized water. My experience is better dilute that in two steps, e.g., first 1:10, and then 1:100.

  2. Sonicate the beads for a few minutes (1-5 mins).

  3. Put bead solution on each coverslip (~25uL for 5mm in diameter coverslip) for 20 mins.

  4. Wash the coverslip several times with ddH2O.

For sparse bead sample

  1. Dilute the stock bead solution 1:10^6 with deionized water. My experience is better dilute that in two steps, e.g., first 1:1000, and then another 1:1000.

  2. Sonicate the beads for a few minutes (1-5 mins).

  3. Put 10^-6 bead solution on each coverslip (~50uL for 5mm in diameter coverslip) for just one min.

  4. Wash the coverslip several times with ddH2O.