Method and system for standardizing microscope instruments

• US 8,027,030 B2
• Filed: 01/24/2011
• Issued: 09/27/2011
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Novartis Ag
• Original Assignee: HistoRx, Inc. (NeoGenomics, Inc.)
• Inventors: Christiansen, Jason, Tedeschi, Gregory R., Pinard, Robert, Zerkowski, Maciej P.
• Primary Examiner(s): Toatley; Gregory J
• Assistant Examiner(s): Underwood; Jarreas C
• Application Number:
  US13/012,707
• Publication Number:
  US 20110116087A1
• Time in Patent Office: 246 Days
• Field of Search: 3562371-2416, 3562421-2438, 356426-431, 356600-640
• US Class Current: 356/243.8
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/276 with alternation of sample and standard in optical path
• CPC Class Codes: G01N 21/278 Constitution of standards
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

Methods and apparatus for standardizing quantitative measurements from a microscope system. The process includes a calibration procedure whereby an image of a calibration slide is obtained through the optics of the microscope system. The calibration slide produces a standard response, which can be used to determine a machine intrinsic factor for the particular system. The machine intrinsic factor can be stored for later reference. In use, images are acquired of a target sample and of the excitation light source. The excitation light source sample is obtained using a calibration instrument configured to sample intensity. The calibration instrument has an associated correction factor to compensate its performance to a universally standardized calibration instrument. The machine intrinsic factor, sampled intensity, and calibration instrument correction factor are usable to compensate a quantitative measurement of the target sample in order to normalize the results for comparison with other microscope systems.

Claims

1. A calibration instrument for sampling illumination of an excitation light source of a microscopy system, comprising:

a calibration surface positioned along an optical path to substantially uniformly scatter illumination from the excitation light source toward a detection portion of the microscopy system;

a dichromatic mirror positioned to reflect illumination from the excitation light source along an optical path through an objective toward a target sample and to transmit at least a portion of illumination from the target sample toward a detection portion of the microscopy system, wherein the calibration surface temporarily blocks the optical path between the dichromatic mirror and the objective during calibration and scatters a substantial portion of the reflected excitation light through the dichromatic mirror toward the detector; and

a housing configured to retain the dichromatic mirror and the calibration surface in a fixed relation with respect to each other, wherein the housing is adapted for placement within an interchangeable filter selection mechanism.

2. The calibration instrument of claim 1, wherein the calibration surface is opaque.

3. The calibration instrument of claim 2, wherein the opaque calibration surface comprises filter paper.

4. The calibration instrument of claim 2, wherein the opaque calibration surface comprises a substantially rigid surface.

5. The calibration instrument of claim 4, wherein the substantially rigid surface comprises a ceramic surface.

6. The calibration instrument of claim 1, further comprising at least one neutral filter positioned to attenuate at least one of the illumination from the excitation light source and the illumination from the target sample, and wherein the housing is further configured to retain the dichromatic mirror, the at least one neutral filter, and the calibration element in fixed relation with respect to each other.