Polarization characteristic measuring method and apparatus

• US 6,025,917 A
• Filed: 05/01/1998
• Issued: 02/15/2000
• Est. Priority Date: 09/24/1996
• Status: Expired due to Term

• Current Assignee: Nikon Corporation
• Original Assignee: Laboratory Of Molecular Biophotonics
• Inventors: Suga, Takayuki, Nakano, Yoshitaro, Shiroshita, Masahisa, Toyonaga, Shuji
• Primary Examiner(s): Font, Frank G.
• Assistant Examiner(s): Punnoose, Roy M.
• Application Number:
  US09/068,087
• Time in Patent Office: 655 Days
• Field of Search: 356/364, 356/367, 356/369, 356/372, 356/300, 356/301, 250/225, 359/385, 359/386
• US Class Current: 356/364
• CPC Class Codes: G01J 3/4406 Fluorescence spectrometry
• CPC Class Codes: G01J 3/447 Polarisation spectrometry
• CPC Class Codes: G01J 4/00 Measuring polarisation of light
• CPC Class Codes: G01N 21/6445 Measuring fluorescence polarisation
• CPC Class Codes: G01N 21/65 Raman scattering

Abstract

Provided is a polarization characteristic measuring method and apparatus for accurately measuring a polarization characteristic of fluorescence or Raman-scattered light emitted when a sample is exposed to light. The sample is exposed to excitation light radiated from a pulsed excitation light source and converted to p-polarized light by polarizer and half-wave plate, and photodetectors measure an intensity I_pp of a p-polarized component and an intensity I_ps of an s-polarized component of fluorescence emitted from the sample under irradiation with the excitation light. In similar fashion, the sample is exposed to the excitation light of s-polarized light and the detectors measure an intensity I_sp of a p-polarized component and an intensity I_ss of an s-polarized component of fluorescence emitted from the sample under irradiation. From these measured values, G factor is calculated according to the following equation:

G=[(I.sub.pp ·I.sub.sp)/(I.sub.ps ·I.sub.ss)].sup.1/2

and polarization response correction is effected based on this G factor to obtain the polarization characteristic of fluorescence.

Claims

1. A polarization characteristic measuring method using beam splitting means for reflecting one of a first beam for irradiation of a sample and a second beam emitted from said sample and for transmitting the other, said polarization characteristic measuring method being adapted for guiding said first beam via said beam splitting means onto said sample, thereby effecting irradiation of the sample, and for measuring a polarization characteristic of said second beam emitted from said sample under the irradiation and traveling via said beam splitting means,

said polarization characteristic measuring method comprising:

a first step of measuring an intensity I_pp of a p-polarized component and an intensity I_ps of an s-polarized component of said second beam emitted when said sample is irradiated with said first beam in the form of p-polarized light with respect to said beam splitting means;

a second step of measuring an intensity I_sp of a p-polarized component and an intensity I_ss of an s-polarized component of said second beam emitted when said sample is irradiated with said first beam in the form of s-polarized light with respect to said beam splitting means;

a third step of calculating a polarization response correction factor G according to the following equation;

space="preserve" listing-type="equation">G=[(I.sub.pp ·

I.sub.sp)/(I.sub.ps ·

I.sub.ss)].sup.1/2 ; and

a fourth step of performing polarization response correction based on said polarization response correction factor G to obtain the polarization characteristic of said second beam.

2. The polarization characteristic measuring method according to claim 1, wherein said second step comprises a step of radiating said first beam toward said sample set in the same location as during measurement of said first step.

3. The polarization characteristic measuring method according to claim 1, wherein said first step comprises a step of radiating said first beam toward said sample located at a position making a predetermined rotation angle about a center axis on a predetermined direction, and

wherein said second step comprises a step of radiating said first beam toward said sample located at a position making a rotation angle 90°

different from said position of the predetermined rotation angle about said center axis.

4. The polarization characteristic measuring method according to claim 1, wherein said fourth step comprises a step of obtaining a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.1 (p)=(I.sub.pp -G·

I.sub.ps)/(I.sub.pp +G·

I.sub.ps);

or

space="preserve" listing-type="equation">P.sub.1 (s)=(G·

I.sub.ss -I.sub.sp)/(G·

I.sub.ss +I.sub.sp).

5. The polarization characteristic measuring method according to claim 1, wherein said fourth step comprises a step of obtaining a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.2 (p)=(I.sub.pp -G·

I.sub.ps)/(I.sub.pp +2·

G·

I.sub.ps);

or

space="preserve" listing-type="equation">P.sub.2 (s)=(G·

I.sub.ss -I.sub.sp)/(G·

I.sub.ss +2·

I.sub.sp).

6. The polarization characteristic measuring method according to claim 1, wherein said fourth step comprises a step of obtaining a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.3 (p)=1-G·

(I.sub.ps /I.sub.pp);

or

space="preserve" listing-type="equation">P.sub.3 (s)=G-(I.sub.sp /I.sub.ss).

7. The polarization characteristic measuring method according to claim 1, wherein said fourth step comprises a step of obtaining a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.4 (p)=(I.sub.pp /I.sub.ps)-G;

or

space="preserve" listing-type="equation">P.sub.4 (s)=G·

(I.sub.ss /I.sub.sp)-1.

8. The polarization characteristic measuring method according to claim 1, wherein said first beam is a beam of a wavelength that can undergo multiple photon absorption by said sample and said second beam is a beam emitted on the occasion of said multiple photon absorption.

9. A polarization characteristic measuring apparatus using beam splitting means for reflecting one of a first beam for irradiation of a sample and a second beam emitted from said sample and for transmitting the other, said polarization characteristic measuring apparatus being adapted for guiding said first beam via said beam splitting means onto said sample, thereby effecting irradiation of the sample, and for measuring a polarization characteristic of said second beam emitted from said sample under the irradiation and traveling via said beam splitting means,

said polarization characteristic measuring apparatus comprising:

a light source portion for outputting the first beam of linearly polarized light for irradiation of the sample;

polarized direction rotating means for rotating a polarized direction of said first beam outputted from said light source portion;

detecting means for detecting intensities of respective p-polarized component and s-polarized component of said second beam traveling via said beam splitting means, out of beams emitted when said sample is irradiated with said first beam;

correction factor calculating means for calculating a polarization response correction factor G according to the following equation;

space="preserve" listing-type="equation">G=[(I.sub.pp ·

I.sub.sp)/(I.sub.ps ·

I.sub.ss)].sup.1/2,

based on an intensity I_pp of a p-polarized component and an intensity I_ps of an s-polarized component of said second beam detected by said detecting means when said sample is irradiated with said first beam changed to p-polarized light with respect to said beam splitting means by said polarized direction rotating means and on an intensity I_sp of a p-polarized component and an intensity I_ss of an s-polarized component of said second beam detected by said detecting means when said sample is irradiated with said first beam changed to s-polarized light with respect to said beam splitting means by said polarized direction rotating means; and

polarization characteristic calculating means for performing polarization response correction based on said polarization response correction factor G to obtain a polarization characteristic of said second beam.

10. The polarization characteristic measuring apparatus according to claim 9, wherein said correction factor calculating means calculates said polarization response correction factor G, based on said intensities I_pp, I_ps, I_sp, and I_ss detected by said detecting means, for said sample located in a fixed location.

11. The polarization characteristic measuring apparatus according to claim 9, further comprising sample rotating means for rotating said sample about a center axis on a predetermined direction,

wherein said correction factor calculating means calculates said polarization response correction factor G. based on said intensities I_pp and I_ps detected by said detecting means, for said sample located at a position making a predetermined rotation angle about said center axis and on said intensities I_sp and I_ss detected by said detecting means, for said sample set at a position making a rotation angle 90°

different from said position of the predetermined rotation angle about said center axis by said sample rotating means.

12. The polarization characteristic measuring apparatus according to claim 9, wherein said polarization characteristic calculating means calculates a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.1 (p)=(I.sub.pp -G·

I.sub.ps)/(I.sub.pp +G·

I.sub.ps);

or

space="preserve" listing-type="equation">P.sub.1 (s)=(G·

I.sub.ss -I.sub.sp)/(G·

I.sub.ss +I.sub.sp).

13. The polarization characteristic measuring apparatus according to claim 9, wherein said polarization characteristic calculating means calculates a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.2 (p)=(I.sub.pp -G·

I.sub.ps)/(I.sub.pp +2·

G·

I.sub.ps);

or

space="preserve" listing-type="equation">P.sub.2 (s)=(G·

I.sub.ss -I.sub.sp)/(G·

I.sub.ss +2·

I.sub.sp).

14. The polarization characteristic measuring apparatus according to claim 9, wherein said polarization characteristic calculating means calculates a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.3 (p)=1-G·

(I.sub.ps /I.sub.pp);

or

space="preserve" listing-type="equation">P.sub.3 (s)=G-(I.sub.sp /I.sub.ss).

15. The polarization characteristic measuring apparatus according to claim 9, wherein said polarization characteristic calculating means calculates a polarization characteristic P of said second beam according to either equation below:

space="preserve" listing-type="equation">P.sub.4 (p)=(I.sub.pp /I.sub.ps)-G;

or

space="preserve" listing-type="equation">P.sub.4 (s)=G·

(I.sub.ss /I.sub.sp)-1.

16. The polarization characteristic measuring apparatus according to claim 9, wherein said light source portion outputs as said first beam a beam of a wavelength that can undergo multiple photon absorption by said sample, and said detecting means detects as said second beam a beam emitted on the occasion of said multiple photon absorption.

17. A polarization characteristic measuring apparatus comprising:

a light source portion for outputting a first beam for irradiation of a sample;

first beam splitting means for guiding said first bean onto said sample to effect irradiation of said sample and receiving a second beam emitted from said sample under the irradiation, said first beam splitting means reflecting one of said first beam and said second beam but transmitting the other;

second beam splitting means having a spectral characteristic equal to that of said first beam splitting means, said second beam splitting means having an plane of incidence 90°

different from an plane of incidence of said first beam splitting means for said second beam, said second beam splitting means receiving said second beam outputted from said first beam splitting means;

detecting means for detecting intensities of respective p-polarized component and s-polarized component of said second beam traveling via said second beam splitting means; and

polarization characteristic calculating means for measuring a polarization characteristic of said second beam, based on the intensities of the respective p-polarized component and s-polarized component of said second beam detected by said detecting means.