Sample analyzing method and sample analyzer

Patent metadata

Field	Value
Publication number	US-2016281151-A1
Application number	US-201615075358-A
Country	US
Kind code	A1
Filing date	Mar 21, 2016
Priority date	Mar 27, 2015
Publication date	Sep 29, 2016
Grant date	—

Abstract

A sample analyzing method includes: denaturing DNA by heating a measurement specimen; bleaching the measurement specimen to inhibit autofluorescence from the measurement specimen; binding a fluorescent dye to a test substance in the measurement specimen; and capturing an image of fluorescence originated from the fluorescent dye by irradiating the measurement specimen with light. The DNA denaturation treatment is performed before the bleaching.

First claim

1 . A sample analyzing method comprising:
denaturing DNA by heating a measurement specimen; bleaching the measurement specimen to inhibit autofluorescence from the measurement specimen; labeling a test substance by binding a fluorescent dye to the test substance in the measurement specimen; and capturing an image of fluorescence originated from the fluorescent dye by irradiating the measurement specimen with light, wherein the denaturing of the DNA is performed before the bleaching.

2 . The sample analyzing method according to claim 1 , further comprising activating the measurement specimen,
wherein the activating is performed before the bleaching.

3 . The sample analyzing method according to claim 1 , further comprising deparaffinizing the measurement specimen,
wherein the deparaffinizing is performed before the bleaching.

4 . The sample analyzing method according to claim 1 , wherein the bleaching inhibits the autofluorescence originated from the measurement specimen by irradiating the measurement specimen with light.

5 . The sample analyzing method according to claim 1 , further comprising detecting the test substance by processing the captured image obtained in the capturing of the image.

6 . The sample analyzing method according to claim 1 , wherein
the measurement specimen is a cell taken from a diseased tissue of a breast cancer, and the test substance is HER2 gene.

7 . The sample analyzing method according to claim 6 , wherein the fluorescent dye is Alexa Fluor 647.

8 . The sample analyzing method according to claim 1 , wherein
the fluorescent dye is switchable between an activation condition in which the fluorescence is excited when the fluorescent dye is irradiated with light for excitation and an inactivation condition in which the fluorescence is not excited even when the fluorescent dye is irradiated with the light for excitation, and the capturing of the image includes
quenching the fluorescence of the fluorescent dye,
activating part of the fluorescent dye whose fluorescence is quenched, and
capturing an image of the fluorescence by irradiating the measurement specimen with the light for excitation.

9 . A sample analyzer comprising:
a pretreatment section that pretreats a measurement specimen before image capture of a test substance included in the measurement specimen; an image capture section that captures an image of the measurement specimen pretreated by the pretreatment section; and an analysis section that extracts the test substance by processing the captured image obtained by the image capture section, wherein the pretreatment section includes
a first treatment unit that performs a DNA denaturation treatment to denature DNA by heating the measurement specimen,
a second treatment unit that performs a bleaching treatment to inhibit autofluorescence from the measurement specimen, and
a third treatment unit that performs a labeling treatment to bind a fluorescent dye to the test substance in the measurement specimen, wherein
the first treatment unit performs the DNA denaturation treatment to the measurement specimen before the second treatment unit performs the bleaching treatment, and the image capture section captures an image of fluorescence originated from the fluorescent dye by irradiating the pretreated measurement specimen with light.

10 . The sample analyzer according to claim 9 , further comprising a fourth treatment unit that performs an activation treatment on the measurement specimen,
wherein the fourth treatment unit performs the activation treatment to the measurement specimen before the second treatment unit that performs the bleaching treatment.

11 . The sample analyzer according to claim 9 , further comprising a fifth treatment unit that performs a deparaffinization treatment on the measurement specimen,
wherein the fifth treatment unit performs the deparaffinization treatment to the measurement specimen before the second treatment unit that performs the bleaching treatment.

12 . The sample analyzer according to claim 9 , wherein the second treatment unit that performs the bleaching treatment inhibits the autofluorescence from the measurement specimen by irradiating the measurement specimen with light.

13 . The sample analyzer according to claim 9 , wherein
the measurement specimen is a cell taken from a diseased tissue of a breast cancer, and the test substance is HER2 gene.

14 . The sample analyzer according to claim 13 , wherein the fluorescent dye is Alexa Fluor 647.

15 . The sample analyzer according to claim 9 , wherein
the fluorescent dye is switchable between an activation condition in which the fluorescence is excited when the fluorescent dye is irradiated with light for excitation and an inactivation condition in which the fluorescence is not excited even when the fluorescent dye is irradiated with the light for excitation, and the image capture section performs
an inactivation process to quench the fluorescence of the fluorescent dye,
an activation process to activate part of the fluorescent dye whose fluorescence is quenched, and
an image capture process to capture an image of the fluorescence by irradiating the measurement specimen with the light for excitation.

Assignees

• Sysmex Corp

Inventors

• Kubo Takuya
• Iwanaga Shigeki

Classifications

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G01N2021/6441

  with two or more labels · CPC title

• G01N1/44

  Sample treatment involving radiation, e.g. heat · CPC title

• C12Q1/6841 Primary

  In situ hybridisation · CPC title

• G01N21/6428 Primary

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title