A simple method to overcome the interference of hemoglobin in the detection of reporter genes in vivo.
The determination of chemiluminescent intensity of reporter gene expression in vivo is generally disturbed by the presence of hemoglobin. Current methods consist in using perfusion to eliminate blood from investigated tumors or organs. In this work we propose a simple method to overcome this difficulty. The method consists in establishing an absorbance-dependence plot of the ratio R% = phi/phi(0) between the chemiluminescent intensities measured when hemoglobin is present or absent. For every measurement of the luminescent intensity phi on sample containing blood, if the absorbance A of the hemoglobin is determined, it allows one to have the intensity ratio R% which in turn gives the corrected intensity phi(0) when the absorption by hemoglobin is eliminated. The method is particularly adapted for comparative measurements of transfection levels in tumors where perfusion cannot be easily performed.
Reynier
Philippe
P
Laboratoire de Chimie Structurale et Spectroscopie Biomoléculaire, UFR de Médecine, CNRS UPRES-A 7031, Université Paris XIII, 74 rue Marcel Cachin, Bobigny Cedex, F93017, France.
Lesage
Denis
D
Cao
An
A
Briane
Dominique
D
Coudert
Robert
R
Taillandier
Eliane
E
D000818:Animals; D002138:Calibration / Q000592:standards; D017930:Genes, Reporter / Q000187:drug effects; D006454:Hemoglobins / Q000737:chemistry / Q000494:pharmacology*; D008081:Liposomes / Q000378:metabolism; D008156:Luciferases / Q000032:analysis*; D008163:Luminescent Measurements; D008545:Melanoma; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured
D006454:Hemoglobins; D008081:Liposomes; D008156:Luciferases