To study the interaction between actin filaments and focal adhesion points in fixed fibroblast cells using a confocal microscope, a researcher plans to perform immunostaining with antibodies against actin and paxillin (a focal adhesion protein). After incubating with the primary antibodies and washing, the researcher plans to use secondary antibodies that bind to the primary antibodies. Below is a list of secondary antibodies conjugated to different fluorophores (dyes) available to the researcher.

A. Alexa 594

B. TRITC

C. Alexa 488

D. Cy5

Which combination of fluorophores should the researcher use to observe co-localization in a confocal microscope?

1
 Alexa 594 (A) and Alexa 488 (C)
2
Alexa 594 (A) and TRITC (B)
3
TRITC (B) and Cy5 (D)
4
Alexa 488 (C) and Cy5 (D)
5
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