Differential fluorescence of the chromocenters and nucleolar equivalents of the yeast nucleus in acridine orange

The nucleus with its limiting membrane and organelles was visible in the majority of the yeast cells stained vitally with the fluorochrome, acridine orange, at a dilution of 1 in 40,000. The intra-nuclear structures could be distinguished by their differential fluorescence. The chromocenters were green while the nucleolar equivalents were orange. The vacuole showed no fluorescence.

Acridine orange leukocyte fluorography in mice

Simultaneous non-invasive visualization of blood vessels and nerves in patients can be obtained in the eye. The retinal vasculature is a target of many retinopathies. Inflammation, readily manifest by leukocyte adhesion to the endothelial lining, is a key pathophysiological mechanism of many retinopathies, making it a valuable and ubiquitous target for disease research. Leukocyte fluorography has been extensively used in the past twenty years; however, fluorescent markers, visualization techniques, and recording methods have differed between studies. The lack of detailed protocol papers regarding leukocyte fluorography, coupled with lack of uniformity between studies, has led to a paucity of standards for leukocyte transit (velocity, adherence, extravasation) in the retina. Here, we give a detailed description of a convenient method using acridine orange (AO) and a commercially available scanning laser ophthalmoscope (SLO, HRA-OCT Spectralis) to view leukocyte behavior in the mouse retina. Normal mice are compared to mice with acute and chronic inflammation. This method can be readily adopted in many research labs.