Near infrared chemical imaging microscope
A chemical imaging system is provided which uses a near infrared radiation microscope. The system includes an illumination source which illuminates an area of a sample using light in the near infrared radiation wavelength and light in the visible wavelength. A multitude of spatially resolved spectra of transmitted, reflected, emitted or scattered near infrared wavelength radiation light from the illuminated area of the sample is collected and a collimated beam is produced therefrom. A near infrared imaging spectrometer is provided for selecting a near infrared radiation image of the collimated beam. The filtered images are collected by a detector for further processing. The visible wavelength light from the illuminated area of the sample is simultaneously detected providing for the simultaneous visible and near infrared chemical imaging analysis of the sample. Two efficient means for performing three dimensional near infrared chemical imaging microscopy are provided.
1. A method for producing a volumetric image of a sample comprising the steps of:
A) illuminating an area of a sample using light in the near infrared wavelength radiation and the visible wavelength radiation so as to yield a near infrared wavelength image and a visible wavelength image of said sample;
B) collecting a spectrum of near infrared wavelength radiation light transmitted, reflected, or emitted from said illuminated area of said sample and producing a collimated beam therefrom;
C) selecting a near infrared wavelength image of said collimated beam with a near infrared imaging spectrometer;
D) capturing said filtered near infrared wavelength image;
E) processing the captured near infrared wavelength image and visible wavelength image;
F) moving said sample relative to an objective;
G) collecting near infrared wavelength and visible wavelength images of said sample relative to said objective in a plurality of focus depths using steps A–F; and
H) processing said collected near infrared wavelength and visible wavelength images to reconstruct said volumetric image of said sample.