Telecentric, wide-field fluorescence scanning systems and methods
Wide-field fluorescence imaging systems and methods. A bi-telecentric optical imaging system comprising imaging optics arranged and positioned such that a first telecentric space is created or exists between a sample platform and an entry aperture stop wherein Principal or chief rays from a plurality of field points on the sample platform are parallel to each other when passing through a first filter; and such that a second telecentric space is created or exists between a light detector and an exit aperture stop wherein the Principal or chief rays from the plurality of field points are parallel to each other when passing through a second filter. In this manner, light collected from different points in the field of view pass through the first filter at the same angles and also through the second filter at the same angles to thereby reduce or eliminate angular spectral shifting effects.
1. A fluorescence imaging system, comprising:
a source subsystem having:
a sample platform holding a fluorescent material;
a light source that illuminates the fluorescent material with excitation light in an absorption band of the fluorescent material; and
a first filter that passes wavelengths of light other than the excitation light, the first filter being positioned in a first light path between the sample platform and an entry aperture stop of a bi-telecentric optical imaging system;
a detector subsystem for detecting light from the fluorescent material, comprising;
a light detector having an array of sensing locations; and
a second filter that passes wavelengths of light in an emission band of the fluorescent material, the second filter being positioned in a second light path between the light detector and an exit aperture stop of the bi-telecentric optical imaging system; and
the bi-telecentric optical imaging system comprising imaging optics arranged and positioned such that a first telecentric space exists in the first light path between the sample platform and the entry aperture stop wherein Principal rays from a plurality of field points on the sample platform are parallel to each other when passing through the first filter; and such that a second telecentric space exists in the second light path between the light detector and the exit aperture stop wherein the Principal rays from the plurality of field points are parallel to each other when passing through the second filter,
wherein contiguous field points on the sample platform are simultaneously imaged onto contiguous sensing locations on the light detector.
2. The imaging system of claim 1 , wherein the bi-telecentric optical imaging system comprises an Offner relay mirror system arrangement comprising a first mirror element having a spherical mirror surface and a second mirror element having a spherical mirror surface, wherein the entry aperture stop and the exit aperture stop each comprise a portion of the first mirror element.
3. The imaging system of claim 2 , wherein the first mirror element presents a convex-shaped mirror surface, and wherein the second mirror element presents a concave-shaped mirror surface.
4. The imaging system of claim 1 , wherein the bi-telecentric optical imaging system comprises a bi-telecentric lens arrangement, wherein the entry aperture stop includes a first refractive lens element and wherein the exit aperture stop includes a second refractive lens element.
5. The imaging system of claim 1 , wherein the light source emits a beam of uniform illumination having a line-shaped profile when impinging on the fluorescent material.
6. The imaging system of claim 1 , wherein the light source comprises a diode laser.
7. The imaging system of claim 1 , wherein the light detector comprises a CCD array detector.
8. The imaging system of claim 7 , wherein the contiguous field points on the sample platform are simultaneously imaged onto contiguous sensing locations on the CCD array detector.
9. A method of imaging a fluorescent material that absorbs light in an absorption band of wavelengths and that emits fluorescent light in an emission band of wavelengths, the method comprising:
a) illuminating a first portion of a fluorescent material on a sample platform with an illumination beam having excitation light in the absorption band; and
b) detecting emissions from the first portion of fluorescent material using a detector system including a light detector having an array of sensing locations, a bi-telecentric optical imaging system, a first filter that passes wavelengths of light other than the excitation light, the first filter being positioned in a first light path between the sample platform and an entry aperture stop of the bi-telecentric optical imaging system, and a second filter that passes wavelengths of light in the emission band, the second filter being positioned in a second light path between the light detector and an exit aperture stop of the bi-telecentric optical imaging system,
wherein the bi-telecentric optical imaging system includes imaging optics arranged and positioned such that a first telecentric space exists in the first light path between the sample platform and the entry aperture stop wherein Principal rays from a plurality of field points on the sample platform are parallel to each other when passing through the first filter; and such that a second telecentric space exists in the second light path between the light detector and the exit aperture stop wherein the Principal rays from the plurality of field points are parallel to each other when passing through the second filter, and
wherein contiguous field points on the sample platform are simultaneously imaged onto contiguous sensing locations on the detector to form a first detector image.
10. The method of claim 9 , further including moving one or both of the sample platform and the illumination beam in a direction substantially parallel with a surface defined by the sample platform so as to illuminate a second portion of the fluorescent material, and repeating the step of detecting to form a second detector image.
11. The method of claim 10 , further including combining the first and second detector images.
12. The method of claim 9 , further including repeatedly moving one or both of the sample platform and the illumination beam in a direction substantially parallel with a surface defined by the sample platform so as to illuminate a multiple portions of the fluorescent material, and repeating the step of detecting to form multiple second detector images, and combining the first detector images and the multiple second detector images.
13. The method of claim 9 , wherein illuminating includes shaping an output of a laser beam so that the beam has a line-shaped profile where it impinges on the sample platform.
14. The method of claim 9 , wherein the bi-telecentric optical imaging system comprises an Offner relay mirror system arrangement including a first mirror element having a spherical mirror surface and a second mirror element having a spherical mirror surface, wherein the entry aperture stop and the exit aperture stop each comprise a portion of the first mirror element.
15. The method of claim 9 , wherein the bi-telecentric optical imaging system comprises a bi-telecentric lens arrangement, wherein the entry aperture stop includes a first refractive lens element and wherein the exit aperture stop includes a second refractive lens element.
16. The method of claim 9 , wherein the light detector comprises a CCD array detector.