Fluorescence

• ProLong is good for Alexa dyes – can affect autofluorescentproteinsEmbedding Media• Note: refractive index of media should be as close to that ofglass and immersion medium• Some media change RI when hardening - Mowiol• Basis: water, glycerine (RI:1,41), polyvinyl alcohol• Keep as this as possible – avoid air bubbles (scattering!)• Fingernail polish as sealant may quench GFP• Don‘t use VectaShield or ProLong with quantum dotsquenching• Antifade agent p-phenylenediamine (Vectashield) quenchescyanine dyes (Cy2, Cy3..)• Crystal Mount quenches some red dyes (phycoerythrin,phycocyanine)