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Quenching fluorescence signal

MD Matthew S. Dietz
TS Thomas L. Sutton
BW Brett S. Walker
CG Charles E. Gast
LZ Luai Zarour
SS Sidharth K. Sengupta
JS John R. Swain
JE Jennifer Eng
MP Michael Parappilly
KL Kristen Limbach
AS Ariana Sattler
EB Erik Burlingame
YC Yuki Chin
AG Austin Gower
JM Jose L. Montoya Mira
AS Ajay Sapre
YC Yu-Jui Chiu
DC Daniel R. Clayburgh
SP SuEllen J. Pommier
JC Jeremy P. Cetnar
JF Jared M. Fischer
JJ Jerry J. Jaboin
RP Rodney F. Pommier
BS Brett C. Sheppard
VT V. Liana Tsikitis
AS Alison H. Skalet
SM Skye C. Mayo
CL Charles D. Lopez
JG Joe W. Gray
GM Gordon B. Mills
ZM Zahi Mitri
YC Young Hwan Chang
KC Koei Chin
MW Melissa H. Wong
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After successful scanning, slides were soaked in 1 × PBS for 10–30 min in a glass Coplin jar, waiting until glass coverslip slid off without agitation. Quenching solution containing 20 mM sodium hydroxide (NaOH) and 3% hydrogen peroxide (H2O2) in 1 × PBS was freshly prepared from stock solutions of 5 M NaOH and 30% H2O2, and each slide placed in 10 mL quenching solution. Slides were quenched under incandescent light, for 30 min for FFPE tissue slides and 20 min for PBMCs adhered to glass slides. Slides were then removed from chamber with forceps and washed three times for two minutes in 1 × PBS. The next round of primary antibodies was applied, diluted in blocking buffer as previously described, and imaging and quenching were repeated over ten rounds for FFPE tissue slides, and two rounds for PBMCs adhered to glass slides.

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