Abstract: A method for calibrating a flow cytometer or fluorescence microscope and for quantitating binding proteins, such as antibodies. Populations of microbeads are provided which are in equilibrium with a saturating amount of fluorescently-labeled protein. The microbeads may have an incorporated fluorescent dye which has a strong signal in a selected channel of a flow cytometer, but no signal in other channels.

Abstract: A system for rapid microbead calibration of a flow cytometer including a suspension of quantitative fluorescent microbead standards and analytical software. The software is used to take information on the microbead suspension from a flow cytometer and analyze data, smooth curves, calculate new parameters and notify of expiration of the system.

Abstract: A method of setting up a flow cytometer using microbeads fluorescing at a plurality of wavelengths, control cells for each of said wavelengths, certified blank microbeads. The blank microbeads are run and the PMT voltages are adjusted so that the blank microbeads are on scale on the flow cytometer. The control cells are run and the photomultiplier tube voltages are adjusted so that the single population falls in selected channels. The fluorescent microbeads are run on the flow cytometer and the Target Channels for the flow cytometer are determined.

Abstract: A kit of highly uniform size microbead standards for flow cytometer alignment, compensation, and/or calibration, comprising a blank microbead population and/or an auto-fluorescent microbead population, together with two or more series of calibrated microbead populations labeled with fluorescent dye(s) which (i) prior to fluorescent dye(s) labeling, match the fluorescence spectra and fluorescence intensity of the blank and/or autofluorescent microbead population, and (ii) after fluorescent dye(s) labeling, match the fluorescence spectra and fluorescence intensity of fluorescently labeled samples to be measured on the flow cytometer. Also disclosed is a corresponding method to align, compensate, and/or calibrate a flow cytometer so as to make measurements on samples comparable and independent of the specific instrument and instrument settings.

Abstract: A qualitative and quantitative method of determining the fluorescence threshold of an instrument and of determining whether the instrument is sensitive enough to measure the autofluorescence of a sample. The method utilizes non-fluorescent particles such a microbeads which are run on a flow cytometer. The peak channel position of the microbeads is used as the fluorescence threshold.

Abstract: A flow cytometer reference standard containing a highly uniform microbead population, wherein each microbead has the same multiple fluorescent labels as the other microbeads and as the samples to be measured. Target conditions are set for the sample after alignment of the flow cytometer. The microbead reference standard is used to determine peak channels (target channels) for each parameter. The target conditions may be reestablished for later uses of the flow cytometer with the same type of samples by using the microbead reference standard to adjust the flow cytometer so that the microbead peak channels fall in the originally determined target channels.

Abstract: Highly uniform microbeads containing a single fluorescence dye or a mixture of fluorescence dyes, which can be excited over a wide range of the spectrum extending from the ultraviolet to the infrared, and which can be used to align a flow cytometer or fluorescence microscope.

Abstract: A method for calibrating a flow cytometer or fluorescence microscope in terms of number of binding antibodies as a function of fluorescence intensity value measured on the flow cytometer or fluorescence microscope, and subsequent measuring of a sample to which the antibodies are bindable. Also disclosed is a microbead calibration kit for carrying out the calibration method of the invention. The disclosed calibration methodology provides a direct relationship between instrument response and numbers of binding antibodies, independent of the fluorochrome employed to label the samples being measured. The method has utility in monitoring the status of an antigenic cellular condition in which the number of antibodies binding to successively obtained cellular samples is determined, to establish the progressionary character of the antigenic cellular condition, in a host from which the cellular samples are taken.

Abstract: A method and structure for mounting fluorescent samples, e.g., fluorescent microbeads, are based on immobilizing the samples in a gel which preserves the spectral properties of the samples and mounting the gel containing the samples on a slide and beneath a coverslip.