Apparatus and method for increasing collection efficiency in capillary based flowcytometry
View Patent ↗In a particle analyzing apparatus including a capillary for passing through a fluid containing particles to be analyzed, an optical system is employed to collect fluorescent light emitted from particles or substances labeled to the particles with improved collection efficiency preserving resolution of the instrument. The optical system may include a first collection lens attached to the capillary and a first reflection element arranged adjacent to the first collection lens configured to reflect fluorescent light of one or more wavelengths. The optical system may include a second collection lens attached to the capillary and a second reflection element arranged adjacent to the second collection lens configured to reflect fluorescent light of one or more wavelengths.
1. A particle analyzing apparatus comprising:
a capillary configured to pass a fluid containing particles to be analyzed, said particles fluoresce or are labeled with substances that fluoresce upon illumination of light;
a source of light;
a first optical system configured to direct a beam of light through the capillary to define an analyzing region in capillary;
a second optical system configured to collect fluorescent light emitted from a particle or substance labeled to the particle in the analyzing region, wherein the second optical system has an optical axis, and comprises a first collection lens attached to a first side of the capillary adjacent to the analyzing region, and a first reflection element arranged adjacent to the first collection lens configured to reflect back fluorescent light of one or more wavelengths in a direction parallel with the optical axis and transmit fluorescent light of wavelength(s) that is (are) different from the one or more wavelengths reflected by the first reflection element; and
a first detector configured to detect fluorescent light transmitted through the first reflection element.
2. The particle analyzing apparatus of claim 1 wherein
the second optical system further comprises a second collection lens attached to a second side of the capillary opposite to the first side, and a second reflection element arranged adjacent to the second collection lens and configured to reflect back fluorescent light of one or more wavelengths in a direction parallel with the optical axis and transmit fluorescent light of wavelength(s) that is (are) different from the one or more wavelengths reflected by the second reflection element; and
the particle analyzing apparatus further comprising a second detector configured to detect fluorescent light transmitted through the second reflection element.
3. The particle analyzing apparatus of claim 2 wherein:
the first reflection element is configured to reflect back fluorescent light of one or more wavelengths that are transmittable by the second reflection element, and transmit fluorescent light of wavelength(s) that are reflectible by the second reflection element; and
the second reflection element is configured to reflect back fluorescent light of one or more wavelengths that are transmittable by the first reflection element, and transmit fluorescent light of wavelength(s) that are reflectible by the first reflection element.
4. A particle analyzing apparatus comprising:
a capillary configured to pass a fluid containing particles to be analyzed, said particles fluoresce or are labeled with substances that fluoresce upon illumination of light;
a source of light;
a first optical system configured to direct a beam of light through the capillary to define an analyzing region in capillary;
a second optical system configured to collect fluorescent light emitted from a particle or substance labeled to the particle in the analyzing region, wherein the second optical system comprises:
a first collection lens at a first side of the capillary adjacent to the analyzing region, and a first reflection element arranged adjacent to the first collection lens configured to reflect fluorescent light of one or more wavelengths and transmit fluorescent light of wavelength(s) that is (are) different from the one or more wavelengths reflected by the first reflection element;
a second collection lens at a second side of the capillary opposite to the first side, and a second reflection element arranged adjacent to the second collection lens and configured to reflect fluorescent light of one or more wavelengths and transmit fluorescent light of wavelength(s) that is (are) different from the one or more wavelengths reflected by the second reflection element;
the first reflection element is configured to reflect fluorescent light of one or more wavelengths that are transmittable by the second reflection element, and transmit fluorescent light of wavelength(s) that are reflectible by the second reflection element; and
the second reflection element is configured to reflect fluorescent light of one or more wavelengths that are transmittable by the first reflection element, and transmit fluorescent light of wavelength(s) that are reflectible by the first reflection element;
a first detector configured to detect fluorescent light transmitted through the first reflection element;
a second detector configured to detect fluorescent light transmitted through the second reflection element;
wherein the second optical system further comprises a first collimating lens arranged between the first collection lens and the first reflection element, and a second collimating lens arranged between the second collection lens and the second reflection element.
5. The particle analyzing apparatus of claim 3 wherein
the second optical system further comprises a third reflection element arranged adjacent to the first reflection element and configured to reflect fluorescent light of a wavelength transmitted from the first reflection element in a direction that is unparallel with the optical axis and transmit fluorescent light of a wavelength transmitted from the first reflection element; and
the particle analyzing apparatus further comprises a third detector configured to detect fluorescent light reflected by the third reflection element.
6. The particle analyzing apparatus of claim 5 wherein the third reflection element is a dichroic filter and placed at an oblique angle with respect to an optical axis.
7. A particle analyzing apparatus of claim 1 comprising:
a capillary configured to pass a fluid containing particles to be analyzed, said particles fluoresce or are labeled with substances that fluoresce upon illumination of light;
a source of light;
a first optical system configured to direct a beam of light through the capillary to define an analyzing region in capillary;
a second optical system configured to collect fluorescent light emitted from a particle or substance labeled to the particle in the analyzing region, wherein the second optical system has an optical axis, and comprises a first collection lens attached to a first side of the capillary adjacent to the analyzing region, and a first reflection element arranged adjacent to the first collection lens configured to reflect fluorescent light of one or more wavelengths and transmit fluorescent light of wavelength(s) that is (are) different from the one or more wavelengths reflected by the first reflection element; and
a first detector configured to detect fluorescent light transmitted through the first reflection element; wherein
the second optical system further comprises a second collection lens attached to a second side of the capillary opposite to the first side, and a second reflection element arranged adjacent to the second collection lens, the second reflection element being a mirror configured to back-reflect fluorescent light of wavelengths of a range collected by the second collection lens in a direction parallel with the optical axis; and
the first reflection element is configured to reflect fluorescent light in a direction unparallel with the optical axis; and
the particle analyzing apparatus further comprises a second detector configured to detect the fluorescent light reflected by the first reflection element.
8. The particle analyzing apparatus of claim 7 wherein the first reflection element is a dichroic filter.
9. The particle analyzing apparatus of claim 1 further comprising a fluidic system configured to transport the fluid containing particles to be analyzed through the analyzing region in the capillary.
10. The particle analyzing apparatus of claim 1 further comprising a data acquisition and analyzing system configured to analyze signals of light detected by the first detector.