title: Improving slide-based assays by stirring: application of liquid-on-liquid mixing to immunofluorescence staining of polytene chromosomes.
abstract: A new method for stirring thin liquid films has been developed and demonstrated to increase the sensitivity of immunofluorescence staining of polytene chromosomes. This liquid-on-liquid mixing (LOLM) technique uses a stirrer fluid, immiscible with the thin film, to transmit shear at the liquid-liquid interface. Here, we stir mineral oil layered over an aqueous thin film of antibody solution, which stains transcription apparatuses on chromosomes previously fixed to a glass slide. The quality of staining was assessed at varying antibody concentrations and incubation or stirring times. Our data indicate that the LOLM technique overcomes the diffusion barrier associated with traditional slide-based biological assays.
authors: Laboratory of Atomic and Solid-State Physics, Cornell University, Ithaca, NY 14853-2501, USA.
authors: Richard C
authors: RC
authors: Yeh
authors: Jerome K
authors: JK
authors: Hyun
authors: Amber K
authors: AK
authors: Boehm
authors: John T
authors: JT
authors: Lis
authors: Carl
authors: C
authors: Franck
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D002875:Chromosomes / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000737:chemistry; D005455:Fluorescent Antibody Technique / Q000295:instrumentation / Q000379:methods; D005898:Glass; D008856:Microscopy, Fluorescence / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D000906:Antibodies