Irradiation unit for a flow-cytometry-based analytical instrument and analytical instrument including the same
View Patent ↗Disclosed is an irradiation unit for a flow-cytometry-based analytical instrument and analytical instrument including the same. The irradiation unit includes a light source, a light beam focusing module for focusing the illuminating light beam on an irradiated area, and a light beam shaper disposed between the light source and the light beam focusing module. The light shaper is used for flattening the distribution of light intensity on the irradiated area in a third direction X that is perpendicular to both a first direction in which the light beam spreads and a second direction in which cells to be detected flow. The irradiation unit and the analytical instrument including the same enable the illuminating light beam to form the irradiated area with the uniform distribution of intensity in a predetermined direction to avoid misjudging particles of the same kind entering into different positions of the irradiated area as different, thereby improving accuracy.
1. An irradiation unit for a flow-cytometry-based analytical instrument, comprising:
a light source for emitting an illuminating light beam in a first direction Z;
a light beam focusing module for focusing the illuminating light beam on an irradiated area, wherein
the irradiated area comprises an area through which cells to be detected flow in a second direction Y; and
a light beam shaper disposed between the light source and the light beam focusing module and configured for flattening distribution of light intensity on the irradiated area along the second direction Y, in which the cells to be detected flow, wherein
the light beam shaper is configured for flattening the distribution of the light intensity along the second direction Y but not along a third direction X that is perpendicular to the first direction Z and the second direction Y, and
the light beam shaper is further configured for broadening the distribution of the light intensity in the second direction Y to expand a size of the irradiation area in the second direction Y to become larger than an original size of an original irradiation area that is irradiated by the light beam without the light beam shaper.
2. The irradiation unit according to claim 1 , wherein
the light source comprises a laser source, which comprises at least two lasers emitting Gauss light beams with different characteristics; and
the light beam shaper is a diffractive optical element.
3. The irradiation unit according to claim 2 , wherein
the diffractive optical element comprises a one-dimensional diffractive optical element for flattening the distribution of light intensity on the irradiated area along the second direction Y in which the cells flow, in which the second direction Y is perpendicular to the first direction Z and the third direction X.
4. The irradiation unit according to claim 1 , wherein a factor β of the light beam shaper satisfies the following formula:
β
=
2
2
π
ω
0
y
0
λ
d
≥
40
,
in which ω 0 is the waist radius of Gauss light beams; y 0 is the half width of a shaped spot; λ is the wavelength of the incident light beam; and d is a distance from the light beam shaper to a shaping plane.
5. The irradiation unit according to claim 1 , wherein
the light beam focusing module comprises a focusing lens module for focusing the light beam in the second direction Y in which the cells flow, wherein a focus point of the focusing lens module is at a center of the irradiated area, and the focusing lens module comprising at least two cylindrical lenses for compressing and magnifying the light beam in the third direction X that is perpendicular to both the second direction Y in which the cells flow and the first direction Z in which the light beam spreads.
6. The irradiation unit according to claim 1 , wherein
the irradiated area is equal in size to the cell diameter in the second direction Y in which the cells flow.
7. A flow-cytometry-based analytical instrument, comprising:
a light source for emitting an illuminating light beam in a first direction Z;
a light beam focusing module for focusing the illuminating light beam on an irradiated area, wherein the irradiated area comprises an area through which cells to be detected flow in a second direction Y;
a light beam shaper disposed between the light source and the light beam focusing module and configured for flattening distribution of light intensity on the irradiated area along the second direction Y, in which the cells to be detected flow, wherein
the light beam shaper is configured for flattening the distribution of the light intensity along the second direction Y but not along the third direction X, and
the light beam shaper is further configured for broadening the distribution of the light intensity in the second direction Y to expand a size of the irradiation area in the second direction Y to become larger than an original size of an original irradiation area that is irradiated by the light beam without the light beam shaper;
a flow chamber for enabling cell flows encased in a sheath fluid to pass through the irradiated area one by one; and
a photodetection unit for collecting optical information from cells passing through the flow chamber and irradiated by the light beam, and converting the optical information into electrical signals.
8. The analytical instrument according to claim 7 , wherein
the light source comprises a laser source, which comprises at least two lasers emitting Gauss light beams with different characteristics; and
the light beam shaper is a diffractive optical element.
9. The analytical instrument according to claim 8 , wherein
the diffractive optical element comprises a one-dimensional diffractive optical element for flattening the distribution of light intensity on the irradiated area along the second direction Y that is perpendicular to both the third direction X and the first direction Z.
10. The analytical instrument according to claim 7 , wherein
a factor β of the light beam shaper satisfies the following formula:
β
=
2
2
π
ω
0
y
0
λ
d
≥
40
,
in which ω 0 is the waist radius of Gauss light beams; y 0 is the half width of a shaped spot; λ is the wavelength of the incident light beam; and d is a distance from the light beam shaper to a shaping plane.
11. The analytical instrument according to claim 10 , wherein
the light beam focusing module comprises a focusing lens module for focusing the light beam in the second direction Y in which the cells flow, wherein a focus point of the focusing lens module is at a center of the irradiated area, and the focusing lens module comprises at least two cylindrical lenses for compressing and magnifying the light beam in the third direction X that is perpendicular to both the second direction Y in which the cells flow and the first direction Z in which the light beam spreads.
12. The analytical instrument according to claim 11 , wherein
the light beam focusing module forms an irradiated area in the flow chamber, and the irradiated area is equal in size to the cell diameter in the second direction Y in which the cells flow.
13. A method for using an irradiation unit for a flow-cytometry-based analytical instrument, comprising:
using a light source for emitting an illuminating light beam in a first direction Z;
causing a light beam focusing module for focusing the illuminating light beam on an irradiated area, wherein
the irradiated area comprises an area through which cells to be detected flow in a second direction Y; and
flattening, by disposing a light beam shaper between the light source and the light beam module, distribution of light intensity on the irradiated area along the second direction Y, in which the cells to be detected flow, wherein
the act of flattening the distribution of the light intensity flattens the distribution of the light intensity along the second direction Y but not along the first direction X and
the light beam shaper is further configured for broadening the distribution of the light intensity in the second direction Y to expand a size of the irradiation area in the second direction Y to become larger than an original size of an original irradiation area that is irradiated by the light beam without the light beam shaper.
14. The method of claim 13 , wherein the act of causing the light beam focusing module for focusing the illuminating light beam on the irradiated area comprises:
focusing the light beam in the third direction X on the irradiated area; and
focusing the light beam in the second direction Y on the irradiated area.
15. The method of claim 13 , further comprising:
adjusting the light beam focusing module or the light beam shaper to approximate a dimension of the irradiated area to a size of one of the cells to be directed.
16. The method of claim 13 , further comprising:
adjusting the light beam focusing module or the light beam shaper such that a ratio of a light distribution width in the third direction X and a light distribution width in the second direction Y of the irradiated area comprises a range of 20:1 to 40:1.
17. The method of claim 13 , wherein the act of flattening the distribution of the light intensity comprises:
adjusting the light beam focusing module or the light beam shaper to increase a distribution width of the light beam in the third direction X in the irradiated area.
18. The method of claim 13 , wherein the act of flattening the distribution of the light intensity comprises:
adjusting the light beam focusing module or the light beam shaper to decrease a distribution width of the light beam in the second direction Y in the irradiated area.
19. The method of claim 13 , wherein the act of flattening the distribution of the light intensity comprises:
using the light beam shaper to obtain at least one scattering signal and at least one fluorescent signal having substantially same intensity for a plurality of same cells which are to be detected and enter the irradiated area from a plurality of positions.
20. The method of claim 19 , wherein the act of using the light beam shaper comprises:
adjusting a first distribution width in the second direction Y in the irradiated area based at least in part upon a size of a particle to be detected; and
adjusting a second distribution width in the third direction X in the irradiated area to obtain a desired ratio between the first distribution width and the second distribution ratio.