Bio-Rad Laboratories, Inc.

David M Heffelfinger

Franklin R Witney

Craig Van Horn

Chris Cunanan

Rebecca Ann Batterson

Renato Salgado

An apparatus capable of measuring quantities of biological or other types of samples that have been labeled using any of a variety of techniques including fluorescence, radioisotopes, enzyme activated light emitting chemicals, and enzyme activated fluorescent materials is provided. The apparatus allows for either simultaneous or sequential acquisition of signals from multiple sample types. The apparatus is not restricted to a particular source or wavelength of excitation or readout light, nor is the apparatus restricted to a particular emission wavelength. The provided scanner includes a source module that preferably contains an internal laser emitting two different wavelengths of approximately the same intensity. An optional external light source may be coupled to the source module, thus adding further flexibility through the addition of other wavelengths (e.g., V, visible, mid-IR, and IR). The scanner also includes a detection module. Within the detection module are two detectors, thus allowing the simultaneous detection of multiple wavelengths. A bifurcated optical cable is used to transfer the excitation and/or readout light from the source module to the sample and subsequently transfer the emitted and/or scattered light from the sample to the detection module. The scanning stage of the scanner is designed to accommodate a variety of samples, ranging from phosphor screens, gels, and fluorescent samples to microtiter plates. An internal microprocessor is used to control the various aspects of the scanner, preferably including translation stage control, source filters, and detection filters. The internal microprocessor may be coupled to an external computer. The external computer may be used to change the programming of the microprocessor, provide a user interface to the microprocessor, process and store test results, and display sample images.

Multi parameter scanner

A method and apparatus for achieving uniform illumination in an electrophoresis apparatus is provided. Uniform illumination allows quantitative measurements of an electrophoresis gel to be made, thus increasing the information which can be obtained from an electrophoretic analysis. Uniform illumination is achieved by scanning the light source, preferably in a trans-illumination mode, across the sample gel in a direction perpendicular to the axis of the source. The light source is comprised of one or more light bulbs placed in a light tray. Variations in light intensity near the source end portions may be minimized using a variety of techniques including extended light bulbs, filters, reflectors, diffusers, or supplemental sources.

Method and apparatus for correcting illumination non-uniformities

A method and apparatus for correcting non-uniformities in the source and the lens assembly of an electrophoresis apparatus is provided. Assuming a detector with a uniform responsivity, correcting for source and lens non-uniformities allows quantitative measurements of an electrophoresis gel to be made, thus increasing the information which can be obtained from an electrophoretic analysis. The non-uniformities due to the illumination source are characterized by sampling a portion of the source with a linear detector array and creating a correction data file. In order to sample the source, a mirror or beamsplitter is appropriately positioned, for example along the central optical axis of the electrophoresis apparatus. Similarly, a correction data file representing the non-uniformities due to the lens assembly is created using a secondary linear source of known uniformity. After the correction data files are stored, an image of the sample is taken and a sample data file is created. The sample data file can be normalized using the correction data files thereby creating a corrected sample file which can either be displayed or stored for future use.

Method and apparatus for the removal of non-uniformities in an

A method and apparatus for correcting non-uniformities in the lens assembly of an electrophoresis apparatus is provided. Assuming a detector with a uniform responsivity as well as a uniform illumination source, correcting for lens non-uniformities allows accurate quantitative measurements of an electrophoresis gel to be made, thus increasing the information which can be obtained from an electrophoretic analysis. Applying the system, the non-uniformities due to the lens assembly are first characterized for a range of aperture and magnification settings. A look-up table is then created which contains the non-uniformities and/or correction data files for the lens assembly according to the aperture and magnification settings. In order to correct a sample image, the aperture and magnification settings used to obtain the sample image are provided to the system processor. These settings may be automatically obtained by the processor or manually input by the user. After the processor receives the lens settings, it applies the look-up table to determine the corresponding lens non-uniformities as well as the necessary correction file. The sample image is then normalized by dividing the sample image file by the appropriate correction file. Once normalized, the corrected sample image file may either be displayed or stored for later use.

Method and apparatus for correcting lens non-uniformities in an

A method and apparatus of imaging fluorescence in situ hybridization (FISH) is provided. The instrument allows the user to simultaneously acquire images from several different colors. This system, used in conjunction with a combinatorial fluorescence approach, is able to create a FISH karyotype with each chromosome being painted with a different color. The optical system is continuously tunable over the detection wavelengths. In one embodiment of the system the sample is simultaneously irradiated in more than one wavelength band and the detection system uses a common path interferometer to scan through the detection wavelengths.

Fluorescence imaging instrument utilizing fish

A method and apparatus for correcting an image of an electrophoresis gel for lens and detector non-uniformities is provided. The removal of such non-uniformities, in conjunction with a uniform illumination source, allows quantitative measurements of an electrophoresis gel to be made, thus increasing the information which can be obtained from an electrophoretic analysis. Lens and detector non-uniformities are removed by first determining the magnitude of the non-uniformities using a calibration standard. The calibration standard has a uniform emittance, preferably in the same wavelength band as the labeled regions of the electrophoresis gel. By placing the calibration standard in close proximity to the entrance aperture of the lens, the calibration process is relatively insensitive to illumination source non-uniformities. Thus an image of the calibration standard taken with the same lens settings as the unknown provides detailed information on the lens and detector non-uniformities. This information in conjunction with a darkfield image is used to correct the sample image.

Method and apparatus for correcting lens and detector non-uniformities

The invention relates to a method of identifying the individual autosomal and sex chromosomes of a human karyotype through the use of a set of combinatorially labeled oligonucleotide probes each member thereof: (i) having a predetermined label distinguishable from the label of any other member of said set, and (ii) being capable of specifically hybridizing with one predetermined autosomal or sex chromosome of a human karyotype.

Method for conducting multiparametric fluorescence in situ hybridization

A method and apparatus of analyzing samples contained in a microplate is provided. The instrument is capable of measuring fluorescence, luminescence, and/or absorption within multiple locations within a sample well. The instrument is tunable over the excitation and/or detection wavelengths. Neutral density filters are used to extend the sensitivity range of the absorption measuring aspect of the instrument. Due to the wavelength tuning capabilities of the instrument, the spectral dependence of the measured fluorescence, luminescence, and absorption of the materials in question can be analyzed. The combination of a data processor and a look-up table improve the ease of operation of the instrument. Several different formats are available for the output data including creation of a bit map of the sample.

Tunable excitation and/or tunable detection microplate reader

A method and apparatus to detect fluorescence from a sample is provided. The optical system allows the user to continuously tune the wavelengths and bandwidths of the excitation source and the emission detection system. A peaking function automatically peaks the detected fluorescent signal by fine tuning the excitation and emission detection systems. A look-up table allows the fluorescent signal for a specific wavelength to be corrected for the wavelength dependence of the optical train. In one embodiment of the system the sample is simultaneously irradiated in more than one wavelength band, each wavelength band being independently tunable.

Tunable excitation and/or tunable emission fluorescence imaging

A novel fiber optic cable having a bifurcated bundle of optical fibers comprised of emitter fiber(s) and collecting fiber(s), a means for focusing light into the proximal end of the emitter fiber(s), a means for focusing light out of the distal end of the emitter fiber(s), and a means for collecting light in the distal end of the collecting fiber(s) is used to increase the sensitivity and specificity of storage phosphor imaging plate scanning. The novel fiber optic cable is used in a storage phosphor imaging plate scanner to transmit light from a light source to the phosphors on the storage phosphor imaging plate and to collect any phosphorescence induced by the light. Use of the bifurcated fiber bundle with the means for focusing light into the emitter fiber(s), focus light out of the emitter fiber(s), and collect light in the collecting fiber(s) provides a method to scan storage phosphor imaging plates with high sensitivity and specificity.

Growing community of inventors

San Pablo, CA, United States of America

David M Heffelfinger