Artifact apparatus to mimic reflection losses of solution-filled microtiter plate readers and related uses thereof
View Patent ↗An apparatus used to calibrate microtiter plate readers. The apparatus includes one or more structures having two exterior surfaces wherein one of the exterior surfaces is coated with a coating selected to establish on that surface a reflection loss that mimics reflection loss at an air-liquid interface, and the other of the two exterior surfaces is coated with a coating selected to establish a reflection loss on that surface that mimics the reflection loss at a bottom surface of a microtiter plate. The apparatus may be a single layer, a multi-layered composition or a container. The apparatus is an artifact that may be used to calibrate a plate reader by mimicking a solution-filled microtiter plate. The artifact may be used for a plurality of liquids, including water. The artifact produces reflection losses more closely mirroring reflection losses expected for a liquid-filled microtiter plate well.
1. An artifact apparatus to aid in the calibration of a microtiter plate reader by mimicking reflection losses, the artifact apparatus comprising a unitary structure having a first side and a second side, wherein the first side includes a first coating that establishes a reflection loss of the first side that mimics reflection loss at an air-liquid interface, wherein the second side includes a second coating that establishes a reflection loss of the second side that mimics the reflection loss at a bottom surface of a microtiter plate; wherein the liquid of the air-liquid interface is water and the first coating enables the first side to reflect about 2% of light incident on it, and wherein the second coating enables the second side to reflect about 6% of light incident on it.
2. The artifact apparatus of claim 1 wherein the unitary structure is a glass structure.
3. The artifact apparatus of claim 1 wherein the first coating includes alternating layers of one or more dielectric materials.
4. The artifact apparatus of claim 1 wherein the second coating includes alternating layers of one or more dielectric materials.
5. An artifact apparatus to aid in the calibration of a microtiter plate reader by mimicking reflection losses, the artifact apparatus comprising:
a. a first component in the form of a first structure, wherein the first component includes a first surface and a second surface;
b. a second component in the form of a second structure, wherein the second component includes a first surface and a second surface, wherein the first structure and the second structure are joined together at their second surfaces, wherein the first surface of the first structure includes a first coating that establishes a reflection loss of the first surface of the first structure that mimics reflection loss at an air-liquid interface, and wherein the second surface of the second structure includes a second coating that establishes a reflection loss of the second surface of the second structure that mimics the reflection loss at a bottom surface of a microtiter plate;
wherein the liquid of the air-liquid interface is water and the first coating enables the first side of the first structure to reflect about 2% of light incident on it; and
wherein the second coating of the second structure enables the second side to reflect about 6% of light incident on it.
6. The artifact apparatus of claim 5 wherein the first structure and the second structure are glass structures.
7. The artifact apparatus of claim 5 further comprising one or more additional components affixed between the first component and the second component.
8. The artifact apparatus of claim 7 wherein one or more of the one or more additional components is a neutral density glass structure.
9. The artifact apparatus of claim 7 wherein one or more of the one or more additional components is an interference filter.
10. The artifact apparatus of claim 5 wherein the first coating includes alternating layers of one or more dielectric materials.
11. The artifact apparatus of claim 5 wherein the second coating includes alternating layers of one or more dielectric materials.
12. An artifact apparatus to aid in the calibration of a microtiter plate reader by mimicking reflection losses, the artifact apparatus comprising a container having a first side and an opposing second side, wherein the first side includes a first coating that establishes a reflection loss of the first side that mimics reflection loss at an air-liquid interface,
wherein the second side includes a second coating that establishes a reflection loss of the second side that mimics the reflection loss at a bottom surface of a microtiter plate, wherein the liquid of the air-liquid interface is water and the first coating enables the first side to reflect about 2% of light incident on it, and wherein the second coating enables the second side to reflect about 6% of light incident on it.
13. The artifact apparatus of claim 12 wherein the first coating includes alternating layers of one or more dielectric materials.
14. The artifact apparatus of claim 12 wherein the second coating includes alternating layers of one or more dielectric materials.
15. A method of the present invention for calibrating a microtiter plate reader using an artifact to mimic reflection losses, the method comprising the steps of:
a. placing an artifact into a calibrated spectrophotometer, wherein the artifact includes a first side and a second side, wherein the first side includes a first coating selected so that the first side is able to establish a reflection loss that mimics reflection loss at an air-liquid interface, wherein the second side includes a second coating selected so that the second side is able to establish a reflection loss that mimics the reflection loss at a bottom surface of a microtiter plate, wherein the second side includes a second coating that establishes a reflection loss of the second side that mimics the reflection loss at a bottom surface of a microtiter plate, wherein the liquid of the air-liquid interface is water and the first coating enables the first side to reflect about 2% of light incident on it, and wherein the second coating enables the second side to reflect about 6% of light incident on it;
b. measuring light absorbance of the artifact in the calibrated spectrophotometer to obtain a calibrated absorbance value for the artifact;
c. placing the artifact in a plate reader under evaluation;
d. measuring the light absorbance of the artifact in the plate reader under evaluation to obtain a measured light absorbance; and
e. comparing the calibrated absorbance value of the artifact with the measured light absorbance of the artifact.
16. The method of claim 15 wherein the artifact is a unitary structure.
17. The method of claim 15 wherein the artifact is a multi-layer structure.
18. The method of claim 15 wherein the artifact is a container.
19. The method of claim 15 wherein the calibrated spectrophotometer is a horizontal or a vertical beam spectrophotometer.