Machine readable storage media for detecting distribution of macromolecules between nucleus and cytoplasm in cells
DC
First Claim
Patent Images
1. A machine readable storage medium comprising a program containing a set of instructions for causing a cell screening system to execute procedures for detecting the distribution of one or more cellular macromolecule of interest between a nucleus and cytoplasm in individual cells, comprising:
- a) scanning multiple cells in an array of locations which contain multiple cells to obtain fluorescent signals from fluorescent reporter molecules in the cells, wherein the cells possess a plurality of fluorescent reporter molecules, wherein the plurality of fluorescent reporter molecules comprises fluorescent reporter molecules to report on;
(i) one or more cellular macromolecule of interest;
(ii) the cell nucleus; and
(iii) the cell cytoplasm;
b) identifying individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules;
c) creating a nuclear mask and a cytoplasmic mask in the individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules;
d) determining an intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest within the nuclear mask and within the cytoplasmic mask in the individual cells in response to contacting the cells with a test stimulus;
e) calculating one or more of the following;
i) a ratio of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
ii) a difference of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
iii) a ratio of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; and
iv) a difference of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
f) comparing the calculations to similar calculations made;
i) in the cells in response to contacting the cells with the test stimulus from at least a second time point; and
/or ii) in the individual cells that have not been contacted with the test stimulus; and
g) determining the effect of the test stimulus on the distribution of the one or more cellular macromolecule of interest between the nucleus and the cytoplasm in the individual cells from the calculations.
3 Assignments
Litigations
0 Petitions
Accused Products
Abstract
The present invention provides systems, methods, and screens for an optical system analysis of cells to rapidly determine the distribution, environment, or activity of fluorescently labeled reporter molecules in cells for the purpose of screening large numbers of compounds for those that specifically affect particular biological functions. The invention involves providing cells containing fluorescent reporter molecules in an array of locations and scanning numerous cells in each location with a high magnification fluorescence optical system, converting the optical information into digital data, and utilizing the digital data to determine the distribution, environment or activity of the fluorescently labeled reporter molecules in the cells. The array of locations may be an industry standard 96 well or 384 well microtiter plate or a microplate which is a microplate having a cells in a micropaterned array of locations. The invention includes apparatus and computerized method for processing, displaying and storing the data.
-
Citations
10 Claims
-
1. A machine readable storage medium comprising a program containing a set of instructions for causing a cell screening system to execute procedures for detecting the distribution of one or more cellular macromolecule of interest between a nucleus and cytoplasm in individual cells, comprising:
-
a) scanning multiple cells in an array of locations which contain multiple cells to obtain fluorescent signals from fluorescent reporter molecules in the cells, wherein the cells possess a plurality of fluorescent reporter molecules, wherein the plurality of fluorescent reporter molecules comprises fluorescent reporter molecules to report on;
(i) one or more cellular macromolecule of interest;
(ii) the cell nucleus; and
(iii) the cell cytoplasm;
b) identifying individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules;
c) creating a nuclear mask and a cytoplasmic mask in the individual cells from the fluorescent signals from the plurality of fluorescent reporter molecules;
d) determining an intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest within the nuclear mask and within the cytoplasmic mask in the individual cells in response to contacting the cells with a test stimulus;
e) calculating one or more of the following;
i) a ratio of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
ii) a difference of average intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
iii) a ratio of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells; and
iv) a difference of integrated intensity of the fluorescent signals from the fluorescent reporter molecules that report on the one or more cellular macromolecule of interest between the nuclear mask and the cytoplasmic mask in the multiple cells;
f) comparing the calculations to similar calculations made;
i) in the cells in response to contacting the cells with the test stimulus from at least a second time point; and
/orii) in the individual cells that have not been contacted with the test stimulus; and
g) determining the effect of the test stimulus on the distribution of the one or more cellular macromolecule of interest between the nucleus and the cytoplasm in the individual cells from the calculations. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
-
Specification
-
- Resources
Litigation Campaign Assessment
Thank you for your request. You will receive a custom alert email when the Litigation Campaign Assessment is available.
×
Litigation Data
-
Current AssigneeCellomics Incorporated (Thermo Fisher Scientific Incorporated)
-
Original AssigneeCellomics Incorporated (Thermo Fisher Scientific Incorporated)
-
InventorsDunlay, R. Terry, Giuliano, Kenneth A., Taylor, D. Lansing, Gough, Albert H.
-
Primary Examiner(s)Marschel, Ardin H.
-
Assistant Examiner(s)Smith, Carolyn L
-
Application NumberUS09/724,376Time in Patent Office1,128 DaysField of Search702/21, 702/19, 382/133, 422/68.1, 435/4US Class Current702/19CPC Class CodesB01J 2219/00315 Microtiter platesB01J 2219/00317 Microwell devices, i.e. hav...B01J 2219/00707 separated from the reactor ...B01J 2219/00743 CellsB82Y 30/00 Nanotechnology for material...B82Y 5/00 Nanobiotechnology or nanome...C12Q 1/68 involving nucleic acidsC12Q 1/6813 Hybridisation assaysC12Q 2525/179 incorporating arbitrary or ...C40B 60/14 for creating librariesG01N 15/1433 using image recognitionG01N 2015/1486 Counting the particlesG01N 2015/1497 Particle shapeG01N 35/00871 Communications between inst...
