Fluorescence microscopy methods and apparatus

Abstract

Methods and apparatus are provided concerning the control of photobleaching of fluorescent labels during the study of samples by fluorescence microscopy. A method is described for operating fluorescence microscopy apparatus to analyse a sample (), the apparatus including input, processing, sample irradiating and detection arrangements (), and the method including the steps of: receiving parameters in the processing arrangement via the input arrangement, wherein the parameters relate to an experiment to be conducted using the apparatus and include at least one parameter relating to a fluorescent label present in the sample; and determining with the processing arrangement an excitation procedure to be carried out during the experiment having regard to the inputted parameters and the rate of photobleaching of the fluorescent label desired during the experiment. Furthermore, confocal fluorescence microscopy apparatus is described which comprises an optical arrangement in the light path from an excitation energy source to a sample which acts to adjust the intensity profile of the light beam across its width so as to be more evenly distributed than a Gaussian profile and/or includes a spinning disk, wherein the rotational speed of the disk is variable under the control of a processing arrangement of the apparatus.