JP2016156708A5 - - Google Patents
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- JP2016156708A5 JP2016156708A5 JP2015034730A JP2015034730A JP2016156708A5 JP 2016156708 A5 JP2016156708 A5 JP 2016156708A5 JP 2015034730 A JP2015034730 A JP 2015034730A JP 2015034730 A JP2015034730 A JP 2015034730A JP 2016156708 A5 JP2016156708 A5 JP 2016156708A5
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- JP
- Japan
- Prior art keywords
- fluorescence
- sample
- fluorescence lifetime
- fluorescent components
- calculating
- Prior art date
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- 238000001514 detection method Methods 0.000 claims 2
- 238000005259 measurement Methods 0.000 claims 2
- 230000000875 corresponding Effects 0.000 claims 1
- 230000003247 decreasing Effects 0.000 claims 1
- 230000005284 excitation Effects 0.000 claims 1
- 230000001678 irradiating Effects 0.000 claims 1
Claims (1)
前記試料が配置される試料室と、
前記試料から放出される蛍光の光強度を検出する検出部と、
前記検出部で検出された光強度の時間変化であるＮ＝１とした第Ｎ減衰曲線を作成して蛍光寿命ｒを算出する制御部とを備える分光蛍光光度計に用いられる含有蛍光成分数決定方法であって、
測定終了時間より所定時間前となる時点から当該測定終了時間までの所定時間間隔となる第Ｘ時間区間の第Ｎ減衰曲線に基づいて、前記試料に含まれる蛍光成分数を１種類と仮定して蛍光寿命ｒＸを算出する第Ｘ蛍光寿命算出ステップと、
前記第Ｘ時間区間より前の所定時間間隔となる第（Ｘ−１）時間区間の第Ｎ減衰曲線に基づいて、前記試料に含まれる蛍光成分数を１種類と仮定して蛍光寿命ｒ（Ｘ−１）を算出するように、当該Ｘ値を減少させながら蛍光寿命ｒ（Ｘ−１）、ｒ（Ｘ−２）・・・を算出していく蛍光寿命変化算出ステップと、
蛍光寿命ｒＸ、ｒ（Ｘ−１）、ｒ（Ｘ−２）・・・が変化しないと判定したときには、前記試料に含まれる蛍光成分数をＮ種類とする判定ステップと、
前記判定ステップで蛍光寿命ｒＸ、ｒ（Ｘ−１）、ｒ（Ｘ−２）・・・が変化していくと判定したときには、蛍光寿命ｒＸの蛍光成分に対応する理論減衰曲線を、第Ｎ減衰曲線から除去することにより、第（Ｎ＋１）減衰曲線を作成する作成ステップと、
第（Ｎ＋１）減衰曲線について、第Ｘ蛍光寿命算出ステップと蛍光寿命変化算出ステップと判定ステップと作成ステップとを実行するように、前記Ｎ値を増加させながら蛍光成分数を算出する多成分算出ステップとを含むことを特徴とする含有蛍光成分数決定方法。 An irradiation unit for irradiating the sample with light having a set excitation wavelength;
A sample chamber in which the sample is disposed;
A detection unit for detecting the intensity of fluorescence emitted from the sample;
Determination of the number of contained fluorescent components used in a spectrofluorometer comprising a control unit that creates an Nth attenuation curve with N = 1, which is a time change in light intensity detected by the detection unit, and calculates a fluorescence lifetime r A method,
Assuming that the number of fluorescent components contained in the sample is one, based on the Nth decay curve in the Xth time interval, which is a predetermined time interval from a time point that is a predetermined time before the measurement end time to the measurement end time. a first X fluorescence lifetime calculating step of calculating a fluorescence lifetime r X,
Based on the Nth decay curve of the (X-1) th time interval, which is a predetermined time interval before the Xth time interval, assuming that the number of fluorescent components contained in the sample is one, the fluorescence lifetime r (X -1) , a fluorescence lifetime change calculating step of calculating fluorescence lifetimes r (X-1) , r (X-2) ... While decreasing the X value;
When it is determined that the fluorescence lifetimes r X , r (X-1) , r (X-2) ... Do not change, a determination step in which the number of fluorescent components contained in the sample is N types;
When it is determined in the determination step that the fluorescence lifetimes r X , r (X-1) , r (X-2) ... Change, a theoretical decay curve corresponding to the fluorescence component of the fluorescence lifetime r X is expressed as follows: Creating a (N + 1) th attenuation curve by removing from the Nth attenuation curve;
A multicomponent calculation step of calculating the number of fluorescent components while increasing the N value so as to execute the Xth fluorescence lifetime calculation step, the fluorescence lifetime change calculation step, the determination step, and the creation step for the (N + 1) th attenuation curve. A method for determining the number of fluorescent components contained.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2015034730A JP6314872B2 (en) | 2015-02-25 | 2015-02-25 | Method for determining the number of fluorescent components contained and a spectrofluorometer using the method for determining the number of fluorescent components contained |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2015034730A JP6314872B2 (en) | 2015-02-25 | 2015-02-25 | Method for determining the number of fluorescent components contained and a spectrofluorometer using the method for determining the number of fluorescent components contained |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2016156708A JP2016156708A (en) | 2016-09-01 |
| JP2016156708A5 true JP2016156708A5 (en) | 2017-07-13 |
| JP6314872B2 JP6314872B2 (en) | 2018-04-25 |
Family
ID=56825782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2015034730A Active JP6314872B2 (en) | 2015-02-25 | 2015-02-25 | Method for determining the number of fluorescent components contained and a spectrofluorometer using the method for determining the number of fluorescent components contained |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP6314872B2 (en) |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4459390B2 (en) * | 2000-06-08 | 2010-04-28 | 浜松ホトニクス株式会社 | Fluorescence measurement method, fluorescence measurement apparatus, and sample evaluation apparatus using the same |
| JP3793531B2 (en) * | 2003-10-07 | 2006-07-05 | オリンパス株式会社 | Fluorescence lifetime measuring device |
| US20060013644A1 (en) * | 2004-07-19 | 2006-01-19 | Eric Yuan | Loose-leaf binder filler with index card function |
| JP2007033159A (en) * | 2005-07-25 | 2007-02-08 | Assay Corp | Measuring object material labeled by fluorescent molecule |
| US7679745B2 (en) * | 2006-11-21 | 2010-03-16 | Neptec Optical Solutions | Time-resolved fluorescence spectrometer for multiple-species analysis |
| US7733470B2 (en) * | 2007-06-26 | 2010-06-08 | Photon Technology International, Inc. | Method and apparatus for real-time measurement and calculation of a fluorescent lifetime |
| JP5437864B2 (en) * | 2010-03-10 | 2014-03-12 | 富士フイルム株式会社 | Operating method of pH measuring device, operating method of detecting device, operating method of biological material analyzing device, and each device |
-
2015
- 2015-02-25 JP JP2015034730A patent/JP6314872B2/en active Active
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