Two excitation wavelength video imaging microscope

• US 5,149,972 A
• Filed: 01/18/1990
• Issued: 09/22/1992
• Est. Priority Date: 01/18/1990
• Status: Expired due to Term

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Fay, Frederic, Dellaville, Fernando
• Primary Examiner(s): Fields, Carolyn E.
• Assistant Examiner(s): Dunn, Drew A.
• Application Number:
  US07/466,889
• Time in Patent Office: 978 Days
• Field of Search: 250/372, 250/461.1, 250/461.2, 250/365
• US Class Current: 250/461.1
• CPC Class Codes: G01J 2001/4493 with image intensifyer tube [IIT]
• CPC Class Codes: G01N 2021/1772 Array detector
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 2201/0696 Pulsed
• CPC Class Codes: G02B 21/0096 with photometer devices photometers per se G01J
• CPC Class Codes: G02B 21/16 adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: H04N 23/70 Circuitry for compensating brightness variation in the scene
• CPC Class Codes: H04N 5/30 Transforming light or analogous information into electric information scanning details H04N3/00; cameras or camera modules comprising electronic image sensors, or control thereof H04N23/00; circuitry of solid-state image sensors [SSIS] or control thereof H04N25/00

Abstract

An imaging apparatus which includes: a fluorescent imaging microscope; a uv radiation source capable of producing a plurality of uv excitation wavelengths; a filter device to select a first and a second excitation wavelength from the plurality of uv excitation wavelengths; a sample chamber to hold a sample for illumination by the radiation of the first and second wavelengths; a photometer to measure the intensity of the excitation wavelength being selected by the filter device and to generate an intensity signal representative of the measured intensity; and a processor in communication with the photometer to record the intensity signal produced by the photometer.

Claims

1. An apparatus for measuring the concentration of an ion in a sample comprising:

a fluorescent imaging microscope;

a uv radiation source capable of producing a plurality of uv excitation wavelengths;

a filter device to select a first and a second excitation wavelength from said plurality of uv excitation wavelengths, said first excitation wavelength capable of exciting fluorescence by said ion in one valence state and said second excitation wavelength capable of exciting fluorescence by said ion in a second valence state;

a sample chamber to hold a sample for illumination by said radiation of said first and second wavelengths;

a photometer to measure the intensity of the excitation wavelength being selected by said filter device and to generate an intensity signal representative of the measured intensity;

a beam splitter to direct a portion of the intensity of said selected wavelength onto said sample to be measured and to direct a second portion of the intensity of said selected wavelength onto said photometer;

optical elements to collect the fluorescent light emitted by said sample;

an image intensifier positioned to receive the fluorescent light collected by said optical elements and to produce a intensified image of said sample;

a video camera to view the intensified image and to produce an electronic signal representative of a frame of said image;

a frame digitizer to digitize said frame of said electronic signal;

a synchronizer in communication with said filter device and said video camera to synchronize the selection of one of said excitation wavelengths with the beginning of the production of said frame of said electronic signal by said video camera; and

a processor in communication with said uv radiation source and said photometer to control the intensity of said uv radiation source and to record the intensity signal produced by said photometer, said computer also in communication with said frame digitizer to process the frame digitized by said frame digitizer and store the processed results on a disk.

2. The apparatus of claim 1 wherein said uv radiation source has a fast rise-time.

3. The apparatus of claim 2 wherein said uv radiation source is a uv strobe.

4. The apparatus of claim 2 wherein said uv radiation source is a uv laser.

5. The apparatus of claim 1 wherein said filter device is a rotatable filter wheel.

6. The apparatus of claim 1 wherein said beam splitter is a dichroic mirror.

7. The apparatus of claim 1 wherein said synchronizer further comprises a filter position sensor to determine which excitation wavelength is being selected and said synchronizer is in communication with said processor.

8. The apparatus of claim 6 wherein said synchronizer comprises a differential amplifier having a first input terminal and a second input terminal, said first input terminal to receive a video synch pulse from said video camera and said second input terminal to receive a position signal from said filter position sensor, said differential amplifier producing an error signal in response thereto.

9. An imaging apparatus comprising:

a fluorescent imaging microscope;

a pulsed uv radiation source capable of producing a plurality of uv excitation wavelengths;

a filter device to select a first and a second excitation wavelength from said plurality of uv excitation wavelengths;

a sample chamber to hold a sample for illumination by said radiation of said first and second wavelengths;

a processor in communication with said radiation source to control the switching on and off of said radiation source in synchrony with the selection of said first and second excitation wavelengths by said filter device.