Table 2.

Solutions and incubation times used for labeling scaffolds.

	Streptavidin-FITC	DBCO-Cy3	fl-HA	Amino-Cy3
Stock	1 mg/ml streptavidin-FITC in PBS (1×) with 0.1% sodium azide	5 mM DBCO-Cy3 in DMSO	0.1 mg/ml fl-HA solution in PBS (1×)	4 mM amino-Cy3 in 20 mM sodium borate buffer
Block	20 min: 0.5% BSA and 0.05% TWEEN in PBS (1×)	1 h: 0.2% TWEEN and 0.2% Triton X-100 in PBS (1×)	Overnight: 0.5% BSA in PBS (1×)	1 h: 0.2% TWEEN and 0.2% Triton X-100 in PBS (1×)
Wash 1	N/A	5 min each: Ultrapure water twice	30 min each: PBS (1×) twice	5 min each: Ultrapure water twice
Label	1 h: Stock diluted to 1:50 with blocking solution	1 h: Stock diluted to 1:100 with PBS with 0.5% BSA	1 h: Stock	30 min: Stock diluted to 0.1 mM in 20 mM sodium borate buffer; add EDC at 0.2 mM and NHS at 0.2 mM
Wash 2	30 min each: PBS (1×) PBS (1×) Ultrapure water	5–10 min each: PBS (1×) Ultrapure water Block solution Ultrapure water 50% IPA 100% IPA 50% IPA Ultrapure water	30 min each: PBS (1×) PBS (1×) PBS (1×) Ultrapure water	5–10 min each: 20 mM sodium borate buffer Ultrapure water Block solution Ultrapure water 50% IPA 100% IPA 50% IPA Ultrapure water