Fluorescence observation apparatus
Fluorescence observation apparatus includes laser scanning fluorescence observation unit and wide field fluorescence observation unit. The former unit includes: lasers; scanning unit that scans sample by laser light; and detector that detects fluorescence from the sample excited by the laser light. The latter unit includes: light source to planar-illuminate the sample; excitation filters that are selectively arranged on light path from the light source, and that correspond to the lasers, respectively; and two-dimensional detector that detects fluorescence from the sample excited by light that has passed through excitation filter arranged on light path from the light source. Each of the excitation filters has optical property in which center wavelength of transmissive band is approximately the same as oscillation wavelength of corresponding laser and bandwidth of transmissive band is approximately the same as tolerance width of oscillation wavelength of corresponding laser.
1. A fluorescence observation apparatus comprising:
a laser scanning fluorescence observation unit including a plurality of lasers, a scanning unit that scans a sample by laser light from the plurality of lasers, and a detector that detects fluorescence from the sample excited by the laser light;
a wide field fluorescence observation unit including a light source to planar-illuminate the sample, a plurality of excitation filters that are selectively arranged on an optical path of light from the light source and that respectively correspond to the plurality of lasers, and a two-dimensional detector that detects a fluorescence from the sample, wherein the sample is excited by light that has passed through an excitation filter arranged on the optical path of light from the light source; and
a dichroic mirror that combines an optical path of the laser light with the optical path of light from the light source;
wherein:
each of the plurality of excitation filters has an optical property in which a center wavelength of a transmissive band is approximately the same as an oscillation wavelength of a corresponding laser and a bandwidth of a transmissive band is approximately the same as a tolerance width of an oscillation wavelength of a corresponding laser;
the laser scanning fluorescence observation unit further includes a plurality of first barrier filters selectively arranged between the dichroic mirror and the detector, each of the first barrier filters corresponding to a respective one of the plurality of lasers;
the wide field fluorescence observation unit further includes a plurality of second barrier filters selectively arranged between the dichroic mirror and the two-dimensional detector, each of the second barrier filters corresponding to a respective one of the plurality of lasers;
the first barrier filter and the second barrier filter which correspond to a same laser have a same optical property; and
when fluorescence observation is performed by using both the laser scanning fluorescence observation unit and the wide field fluorescence observation unit, (i) an excitation filter from the plurality of excitation filters, which corresponds to another laser selected from the plurality of lasers, excluding a laser to be used for fluorescence observation, is arranged on the optical path of light from the light source, (ii) a first barrier filter from the plurality of first barrier filters, which corresponds to the laser to be used for fluorescence observation, is arranged between the dichroic mirror and the detector, and (iii) a second barrier filter from the plurality of second barrier filters, which corresponds to the another laser, is arranged between the dichroic mirror and the two-dimensional detector.
2. The fluorescence observation apparatus according to claim 1 , wherein the light source is a light source with a continuous or discrete light source wavelength including oscillation wavelengths of the plurality of lasers.
3. The fluorescence observation apparatus according to claim 2 , wherein the light source is a mercury lamp.
4. The fluorescence observation apparatus according to claim 2 , wherein the light source is a white LED light source provided with a plurality of LED chips.
5. The fluorescence observation apparatus according to claim 2 , wherein the light source is a white LED light source having a fluorescent material around an LED chip.
6. A fluorescence observation apparatus comprising:
a laser scanning fluorescence observation unit including a plurality of lasers, a scanning unit that scans a sample by laser light from the plurality of lasers, and a detector that detects fluorescence from the sample excited by the laser light;
a wide field fluorescence observation unit including a light source to planar-illuminate the sample, a plurality of excitation filters that are selectively arranged on an optical path of light from the light source and that respectively correspond to the plurality of lasers, and a two-dimensional detector that detects a fluorescence from the sample, wherein the sample is excited by light that has passed through an excitation filter arranged on the optical path of light from the light source;
a dichroic mirror that combines an optical path of the laser light with the optical path of light from the light source; and
a stimulation unit that shares the plurality of lasers with the laser scanning fluorescence observation unit and optically stimulates the sample by using laser light from the plurality of lasers;
wherein:
each of the plurality of excitation filters has an optical property in which a center wavelength of a transmissive band is approximately the same as an oscillation wavelength of a corresponding laser and a bandwidth of a transmissive band is approximately the same as a tolerance width of an oscillation wavelength of a corresponding laser;
the laser scanning fluorescence observation unit further includes a plurality of first barrier filters selectively arranged between the dichroic mirror and the detector, each of the first barrier filters corresponding to a respective one of the plurality of lasers;
the wide field fluorescence observation unit further includes a plurality of second barrier filters selectively arranged between the dichroic mirror and the two-dimensional detector, each of the second barrier filters corresponding to a respective one of the plurality of lasers; and
the first barrier filter and the second barrier filter which correspond to a same laser have a same optical property.
7. The fluorescence observation apparatus according to claim 6 , wherein the light source is a light source with a continuous or discrete light source wavelength including oscillation wavelengths of the plurality of lasers.
8. The fluorescence observation apparatus according to claim 7 , wherein the light source is a mercury lamp.
9. The fluorescence observation apparatus according to claim 7 , wherein the light source is a white LED light source provided with a plurality of LED chips.
10. The fluorescence observation apparatus according to claim 7 , wherein the light source is a white LED light source having a fluorescent material around an LED chip.