Direct immunofluorescence (DIF) is a valuable diagnostic tool in the dermatology clinic. The proper use of a biopsy for DIF is dependent on several factors, including appropriate clinical indication, correct clinical site selection, and proper specimen handling and transport. Improper use of DIF can lead to false negatives, decreased diagnostic yield, and poor resource utilization. This article provides instruction on the appropriate indications and biopsy site selection for DIF. Three examples of skin diseases in which DIF would be particular useful when making a diagnosis are provided.