Charge-coupled device imaging apparatus

Claim 1

An apparatus for imaging of a biological sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional cooled or non-cooled charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for operating said charge-coupled device detector array at, or below, ambient temperature in an on-chip integrating mode under the control of a computer programmed to have the capability of varying signal acquisition time from said charge-coupled device detector array, converting the signal from each pixel of the charge-coupled detector array to a digital value, and storing said digital values as an array of values in memory accessed by said computer, and wherein the computer is programmed to be able to acquire a brightfield correction value for each pixel in the absence of the sample, and to multiply the darkfield-corrected value of the illuminated signal from each pixel by its brightfield correction value prior to readout to the display unit.

Claim 2

An apparatus for imaging of a biological sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional cooled or non-cooled charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for operating said charge-coupled device detector array at, or below, ambient temperature in an on-chip integrating mode under the control of a computer programmed to have the capability of varying signal acquisition time from said charge-coupled device detector array, converting the signal from each pixel of the charge-coupled detector array to a digital value, and storing said digital values as an array of values in memory accessed by said computer, and wherein the computer is programmed to be able to acquire a brightfield correction value for each pixel in the absence of the sample, and to multiply the darkfield-corrected value of the illuminated signal from each pixel by its brightfield correction value prior to readout to the display unit.

Claim 8

An apparatus for light imaging of a translucent sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for correcting the fluorescence value recorded for each pixel to compensate for the variation in incident energy to which that pixel is subjected due to variations in excitation energy reaching different areas of the sample contained in the imaging apparatus.

Claim 9

An apparatus for light imaging of a translucent sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for correcting the fluorescence value recorded for each pixel to compensate for the variation in incident energy to which that pixel is subjected due to variations in excitation energy reaching different areas of the sample contained in the imaging apparatus.

Claim 10

An apparatus for light imaging of a translucent sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for obtaining the image readout from the charge-coupled device detector array through a single channel output.

Claim 11

An apparatus for light imaging of a translucent sample comprising means for illuminating the sample to selectively excite molecules therein and cause fluorescence, a two-dimensional charge-coupled device detector array positioned to detect fluorescent emissions from the sample, means for varying the signal acquisition time from said charge-coupled device detector array so that the integrated signal from each pixel of said charge-coupled device detector array is below its saturation level, means for acquiring a darkfield value for each pixel of said charge-coupled device detector array and subtracting said darkfield value from the illuminated signal from that pixel to yield a darkfield-corrected signal, means for reading out said darkfield-corrected signal to a visual display unit thereby producing a two-dimensional gray scale image of the sample, and means for obtaining the image readout from the charge-coupled device detector array through a single channel output.