Cytological stain composition including verification characteristic

• US 6,661,501 B1
• Filed: 10/29/1999
• Issued: 12/09/2003
• Est. Priority Date: 10/29/1999
• Status: Expired due to Term

• Current Assignee: Cytyc Corporation (Hologic Incorporated)
• Original Assignee: Cytyc Corporation (Hologic Incorporated)
• Inventors: Zahniser, David J., Isenstein, Louise M., Soule, Norman W.
• Primary Examiner(s): Smith, Zandra V.
• Application Number:
  US09/430,196
• Time in Patent Office: 1,502 Days
• Field of Search: 356/39, 356/432, 356/436, 356/437, 436/46, 436/63, 436/164, 436/169, 422/66, 422/68.1, 422/82.09
• US Class Current: 356/39
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1472 with colour
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G01N 21/3151 using two sources of radiation of different wavelengths G01N21/33 - G01N21/39 take precedence
• CPC Class Codes: G06T 2207/30024 Cell structures in vitro; Tissue sections in vitro
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06T 7/70 Determining position or orientation of objects or cameras camera calibration G06T7/80

Abstract

The present invention relates to the analysis of cytological material. Specifically, the invention relates to stains and methods of producing the stains, methods of staining cells for cytological or histological analysis to contrast the nuclear portion of the cell from the cytoplasmic portion, and systems and methods for illuminating a cytological sample. The analysis can be automated or manual.

Claims

1. A method for verifying a cytological stain, the method comprising the steps of:

staining a cytological sample with a stain having a multiple absorption spectrum;

verifying the stain by illuminating the sample with a light source having a first narrow band wavelength that is substantially absorbed by the stain; and

imaging the sample by illuminating the sample with the light source having a second narrow band wavelength that is substantially absorbed by the stain.

2. The method of claim 1, wherein the first narrow band wavelength is between about 690 nm and about 750 nm.

3. The method of claim 1, wherein the light source comprises a red LED.

4. The method of claim 1, wherein the second narrow band wavelength is between about 500 nm and about 600 nm.

5. The method of claim 1 wherein the light source comprises a green LED.

6. The method of claim 1, wherein the multiple absorption spectrum stain absorbs light at two different narrow band wavelengths.

7. The method of claim 1, wherein the light source comprises a first light source and a second light source.

8. The method of claim 7, wherein the first light source comprises a red LED.

9. The method of claim 7, wherein the second light source comprises a green LED.

10. A method for verifying a cytological stain staining a sample, the method comprising the steps of:

illuminating the sample with a light source having a first wavelength band;

verifying the stain used based upon light absorption of the sample; and

imaging the sample by illuminating the sample with a light source having a second wavelength band different than the first wavelength band.

11. The method of claim 10 wherein the stain absorbs light at a narrow wavelength.

12. A method for verifying a cytological stain staining a sample, the method comprising the steps of:

verifying the stain by illuminating the sample with a light source having a first narrow band wavelength that is substantially absorbed by the stain; and

imaging the sample by illuminating the sample with the light source having a second narrow band wavelength that is substantially absorbed by the stain.

13. The method of claim 12 further comprising the step of staining the cytological sample with a stain having a multiple absorption spectrum.

14. The method of claim 12 wherein the light source comprises a first light source and a second light source.

15. The method of claim 14 wherein the first light source comprises a red LED.

16. The method of claim 14 wherein the second light source comprises a green LED.

17. A method of imaging a specimen stained with a stain having multiple spectral absorption peaks, the method comprising the steps of:

illuminating the stained specimen with a multiple wavelength light source; and

obtaining an image of the stained specimen, wherein the image is obtained at a narrow band wavelength generally corresponding to a valley formed between two of the multiple spectral absorption peaks of the stain.

18. The method of claim 17 wherein the image is obtained while illuminating the specimen at at least two wavelengths simultaneously.