ARS | Publication request: Evaluating autofluorescence in live and dead tarnished plant bug (Lygus lineolaris)

Submitted to: National Entomological Society of America Annual Meeting
Publication Type: Other
Publication Acceptance Date: July 9, 2008
Publication Date: November 18, 2008
Citation: Allen, M.L. 2008. Evaluating autofluorescence in live and dead tarnished plant bug (Lygus lineolaris). National Entomological Society of America Annual Meeting, Paper # 35282.

Technical Abstract: Insects exhibit a wide variety of colors, often beautiful, when viewed in daylight conditions. When examined with ultraviolet illumination and fluorescence microscope filters, an additional spectrum of color is visible. Furthermore, some of the fluorescence of insects is visible only in live insects, and entirely different fluorescence is apparent in dead insects. When insects are the subjects of molecular studies, fluorescent protein markers are used to indicate somatic and germline transformation. The most popular of these fluorescent proteins, green fluorescent protein (GFP), is a small (238 amino acid coding sequence) protein used in transformation vectors. Its discovery and development led to the 2008 Nobel Prize in Chemistry. GFP and other fluorescent proteins will be used in molecular studies of Lygus lineolaris. Detection of the fluorescence of the marker protein can be masked by the inherent fluorescence, or autofluorescence, of the study organism. Photodocumentation of GFP and other fluorescent markers often requires long exposure times, which means the specimens must be immobilized or dead when photographed. In preparation for fluorescence studies in L. lineolaris, the autofluorescence characteristics at pertinent stages of development have been examined and documented. In addition, different methods of destructive specimen preparation were examined. Samples were frozen and photographed, and heat-killed and photographed. The observed autofluorescence characteristics will facilitate design of molecular study methods that will detect genetic mechanisms in this important insect pest.