"So you sit down at a microscope expecting to be blinded by colour, but we're met with darkness. So did we just make a massive breakthrough where we discover our protein is absent? Or did we maybe mess up our protocol? This forces us to be really impartial judges and harsh critics for our own data. So take this cell line that I was testing. I didn't know if the protein of interest, CyFib1-2, which associates with translational machinery, would be expressed. In the image we can see a web-like pattern which is very reminiscent of the ER, suggesting that this is a positive hit. If the channel is dark, go back to the red lab, prepare a counterstain and compare it to a cell line where you know the protein is expressed. If you go back and the counterstain works but the channel is still dark, congratulations, you have a new biological result. If both are dark, then you probably need to review your protocol. Good luck!"