Microwave-assisted synthesis of thiophene fluorophores, labeling and multilabeling of monoclonal antibodies, and long lasting staining of fixed cells

J Am Chem Soc. 2009 Aug 12;131(31):10892-900. doi: 10.1021/ja902416s.

Abstract

We report the expedient microwave-assisted synthesis of thiophene based 4-sulfo-2,3,5,6,-tetrafluorophenyl esters whose molecular structure was engineered to achieve blue to red bright fluorescence. The reactivity toward monoclonal antibodies of the newly synthesized fluorophores was analyzed in comparison with that of the corresponding N-succinimidyl esters. Single-fluorophore and multiple-fluorophore labeled antibodies were easily prepared with both types of esters. Multiple-fluorophore labeling with blue and orange emitting fluorophores resulted in white fluorescent antibodies. Thiophene based fluorophores displayed unprecedented fluorescence stability in immunostaining experiments. First-principles TD-DFT theoretical calculations helped us to interpret the behavior of fluorescence emission in different environments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal* / chemistry
  • Cell Line
  • Fluorescent Dyes / chemical synthesis*
  • Humans
  • Lymphocytes
  • Microwaves
  • Staining and Labeling / methods*
  • Thiophenes / chemistry*
  • Tissue Fixation

Substances

  • Antibodies, Monoclonal
  • Fluorescent Dyes
  • Thiophenes