Immunofluorescent staining method for use with monoclonal antibodies

This note describes an immunofluorescent staining method for cells in the S-phase which have been allowed to take up bromodeoxyuridine into their DNA in place of thymine. The technique involves the use of fluorescinated monoclonal antibodies against bromodeoxyuridine and allows rapid and accurate estimation of cells in the S-phase, the technique does not require interpretation by skilled technicians.     

Prevention of Chemography in Prestained Epoxy Autoradiographs by an Intervening Carbon Film

Evaporation of a carbon layer over toluidine blue stained epoxy sections prior to applying emulsion eliminates or significantly reduces chemography that would otherwise be present in autoradiographs. This simple procedure permits routine proceasing of large numbers of slides without the limitations of the impermeable membranes currently recammended for light microscopic autoradiography following prestaining. The method permits “before and after” microphotography and use of simple staining procedures for sections to be studied autoradiographically.     

Proliferating hair cell precursors in the ear of a postembryonic fish are replaced after elimination by cytosine arabinoside

Identified, proliferating S-phase cells in the postembryonic fish ear are known to be the precursors to new hair cells. It is not known, however, whether the ability to proliferate is restricted to a small population of cells. The ability of cells that are not normally in the cell cycle to enter S-phase was examined using the antimitotic drug cytosine arabinoside (ara-C). The normal population of S-phase cells in the saccule was destroyed by a single large dose of ara-C. Two weeks later, the prsence of S-phase cells was evaluated using the S-phase marker bromodeoxyuridine. The results strikingly demonstrate that S-phase cells are replaced, since S-phase cells returned to the saccule in the same number as found in normal fish. The data are interpreted to suggest that a large number of nonsensory support cells are capable of entering the cell cycle and that some mechanism must regulate which of these are actually cycling at any given time. © 1995 John Wiley & Sons, Inc.