Sample carrier and method for imaging a sample
A sample carrier for receiving a sample includes an optical medium in which the sample is received, the optical medium having a first refractive index. A window portion defining two parallel surfaces includes an optically transparent material having a second refractive index, and is arranged at a bottom side of the sample carrier. The first and second refractive indices do not deviate by more than 2.5%.
1 . An imaging system for imaging a sample, comprising:
a sample carrier for receiving the sample, the sample carrier comprising:
an optical medium in which the sample is received, the optical medium having a first refractive index; and
a window portion defining two parallel surfaces that comprises an optically transparent material having a second refractive index, and that is arranged at a bottom side of the sample carrier, wherein the first and second refractive indices do not deviate by more than 2.5%;
a sample carrier receiving portion configured to receive the sample carrier;
an optical detection system, comprising a first objective arranged below, above or to the side of the sample carrier receiving portion and being configured to receive detection light emitted by the sample; and
a second objective arranged below, above or to the side of the sample carrier receiving portion and configured to receive the detection light emitted by the sample,
wherein an optical axis of the first objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion, and
wherein an optical axis of the second objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion.
2 . The imaging system according to claim 1 , wherein the sample carrier comprises at least one of a microplate, a carrier body, a lid or a cover, a sample container, a petri dish, a chamber slide, and a flow cell.
3 . The imaging system according to claim 1 , wherein the parallel surfaces are plan parallel surfaces.
4 . The imaging system according to claim 1 , wherein the optically transparent material is one of the following polymers or their derivatives: BIO-133, FEP, PEG-DA, PTFE, collagen, Matrigel™, agarose, alginate, cellulose, or polyacrylamide; and wherein the second refractive index has a value between 1.2977 and 1.3643.
5 . The imaging system according to claim 1 , wherein the optically transparent material is one of the following polymers or their derivatives: PMMA, EVASKY S87, or Optorez; and wherein the second refractive index has a value between 1.47 and 1.49.
6 . The imaging system according to claim 1 , wherein the optically transparent material is one of the following polymers or their derivatives: pSX-DPM, pMA-DPM, a polycarbonate, a polyphosphonate, or a polyester; and wherein the second refractive index has a value between 1.55 and 1.56.
7 . The imaging system according to claim 1 , wherein the angle enclosed by the optical axis of the at least one objective and the parallel surfaces of the window portion when the sample carrier is received in the sample carrier receiving portion is between 10° and 80°.
8 . The imaging system according to claim 1 , wherein the optical axis of the first objective and the optical axis of the second objective intersect each other; or wherein the first objective and the second objective are arranged relative to each other such that a field of view of the first objective and a field of view of the second objective intersect each other; or wherein the first objective and the second objective are adapted to generate different views of the sample.
9 . The imaging system according to claim 1 , further comprising an illumination system configured to create illumination light and to direct the illumination light through the first objective onto the sample.
10 . The imaging system according to claim 9 , wherein the illumination system is configured to form a light sheet by directing the illumination light through the first objective onto the sample.
11 . The imaging system according to claim 1 , wherein the sample carrier comprises at least one flow cell, the at least one flow cell comprising the window portion; wherein the imaging system comprises a fluid or pneumatic pump device configured to move the optical medium through the at least one flow cell, thereby moving samples received in the optical medium along a flow direction past the window portion, the flow direction being parallel to the parallel surfaces; and wherein the optical detection system is configured to sequentially image the samples being moved past the window portion.
12 . The imaging system according to claim 1 , wherein the sample carrier receiving portion is movable along at least one direction parallel to the parallel surfaces or relative to the first objective.
13 . The imaging system according to claim 1 , wherein the first objective is an immersion objective; wherein the imaging system comprises an immersion medium that is arranged between a front lens of the at least one objective and the window portion of the sample carrier; wherein the immersion medium has a third refractive index; and wherein the first and third refractive indices and/or the second and third refractive indices do not deviate by more than 2.5%.
14 . The imaging system according to claim 1 , wherein the optical detection system is a widefield microscope.
15 . The imaging system according to claim 1 , wherein the imaging system is configured to perform at least one of widefield microscopy, widefield fluorescence microscopy, spinning disc microscopy, fluorescence lifetime microscopy, and light sheet microscopy.
16 . A method for imaging a sample with an imaging system, the method comprising the following steps:
providing a sample carrier having an optical medium for embedding the sample, the optical medium having a first refractive index, and a window portion defining two parallel surfaces, comprising an optically transparent material that has a second refractive index, and is arranged at a bottom side of the sample carrier, wherein the first and second refractive indices do not deviate by more than 2.5%;
arranging the sample in the sample carrier by immersing the sample in the optical medium;
positioning the sample carrier in a sample carrier receiving portion of an imaging device; and
capturing at least one image of the sample with an optical detection system of the imaging device, wherein the optical detection system comprises first objective arranged below, above or to the side of the sample carrier and configured to receive detection light emitted by the sample,
wherein the optical axis of the first objective and the parallel surfaces of the window portion enclose an oblique angle,
wherein the imaging system comprises a second objective arranged below, above or to the side of the sample carrier receiving portion and configured to receive the detection light emitted by the sample, and
wherein the optical axis of the second objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion.
17 . An imaging system for imaging a sample, comprising:
a sample carrier for receiving a sample, comprising:
an optical medium in which the sample is received, the optical medium having a first refractive index; and
a window portion defining two parallel surfaces that comprises an optically transparent material having a second refractive index, and that is arranged at a bottom side of the sample carrier, wherein the first and second refractive indices do not deviate by more than 2.5%;
a sample carrier receiving portion configured to receive the sample carrier;
an optical detection system, comprising a first objective arranged below, above or to the side of the sample carrier receiving portion and being configured to receive detection light emitted by the sample, wherein an optical axis of the first objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion; and
a second objective arranged below, above or to the side of the sample carrier receiving portion and configured to receive the detection the light emitted by the sample, wherein an optical axis of the second objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion,
wherein at least one of an illumination characteristic and a detection characteristic are different for the first objective and for the second objective for generating images from different dyes in the sample with the first and the second objective, or
wherein the optical axis of the first objective encloses an angle with the optical axis of the second objective within a range of 50° to 90°.
18 . The imaging system according to claim 17 , wherein the at least one of the illumination characteristic and the detection characteristic are different for the first objective and for the second objective for generating images from different dyes in the sample with the first and the second objective.
19 . The imaging system according to claim 17 , wherein the optical axis of the first objective encloses the angle with the optical axis of the second objective within the range of 50° to 90°.
20 . A method for imaging a sample with an imaging system, the method comprising the following steps:
providing a sample carrier having an optical medium for embedding the sample, the optical medium having a first refractive index, and a window portion defining two parallel surfaces, comprising an optically transparent material that has a second refractive index, and is arranged at a bottom side of the sample carrier, wherein the first and second refractive indices do not deviate by more than 2.5%;
arranging the sample in the sample carrier by immersing the sample in the optical medium;
positioning the sample carrier in a sample carrier receiving portion of an imaging device; and
capturing at least one image of the sample with an optical detection system of the imaging device, wherein the optical detection system comprises a first objective arranged below, above or to the side of the sample carrier and configured to receive detection light emitted by the sample,
wherein the optical axis of the first objective and the parallel surfaces of the window portion enclose an oblique angle,
wherein the imaging system comprises a second objective arranged below, above or to the side of the sample carrier receiving portion and configured to receive the detection light emitted by the sample, and wherein the optical axis of the second objective and the parallel surfaces of the window portion enclose an oblique angle when the sample carrier is received in the sample carrier receiving portion,
wherein at least one of an illumination characteristic and a detection characteristic are different for the first objective and for the second objective for generating images from different dyes in the sample with the first and the second objective, or
wherein the optical axis of the first objective encloses an angle with the optical axis of the second objective within a range of 50° to 90°.