Optical filter system and fluorescence detection system

Patent metadata

Field	Value
Publication number	US-10145738-B2
Application number	US-201815905052-A
Country	US
Kind code	B2
Filing date	Feb 26, 2018
Priority date	Sep 1, 2015
Publication date	Dec 4, 2018
Grant date	Dec 4, 2018

Abstract

An optical detection filter has a transmission spectrum for detecting fluorescence light of a plurality of different fluorescent dyes. In the range between 350 nm and 1000 nm, the transmission spectrum has a first stopband (DS1) from Dλ1 to Dλ2, a first passband (DD1) from Dλ2 to Dλ3, a second stopband (DS2) from Dλ3 to Dλ4, a second passband (DD2) from Dλ4 to Dλ5, a third stopband (DS3) from Dλ5 to Dλ6, and a third passband (DD3) from Dλ6 to Dλ7. The stopbands (DS1, DS2, DS3) each have a mean transmittance of at most 0.01, typically at most 0.001 or at most 0.0001, and the passbands (DD1, DD2) each have a mean transmittance of at least 0.5, typically at least 0.8 or at least 0.9; wherein 350 nm≤Dλ1<Dλ2<Dλ3<Dλ4<Dλ5<Dλ6<Dλ7<1000 nm.

First claim

The invention claimed is:

1. An optical filter system for observing fluorescence, comprising:
an illumination filter; and
a detection filter;
wherein a transmission spectrum of the illumination filter, in a wavelength range from 350 nm to 1000 nm, has
a first stopband ( B S1) from 350 nm to B λ1 with a mean transmittance of B TS1,
a first passband ( B D1) from B λ1 to B λ2 with a mean transmittance of B TD1,
a second stopband ( B S2) from B λ2 to B λ3 with a mean transmittance of B TS2,
a second passband ( B D2) from B λ3 to B λ4 with a mean transmittance of B TD2, and
a third stopband ( B S3) from B λ4 to BAS with a mean transmittance of B TS3;

wherein 350 nm≤ B λ1< B λ2< B λ3< B λ4< B λ5≤1000 nm;
wherein B TS1< B TD1; B TD1> B TS2; B TS2< B TD2; B TD2> B TS3;
wherein a transmission spectrum of the detection filter, in the wavelength range from 350 nm to 1000 nm, has
a first stopband ( D S1) from 350 nm to D λ1 with a mean transmittance of D TS1,
a first passband ( D D1) from D λ1 to D λ2 with a mean transmittance of D TD1,
a second stopband ( D S2) from D λ2 to D λ3 with a mean transmittance of D TS2,
a second passband ( D D2) from D λ3 to D λ4 with a mean transmittance of D TD2, and
a third stopband ( D S3) from D λ4 to D λ5 with a mean transmittance of D TS3;

wherein 350 nm≤ D λ1< D λ2< D λ3< D λ4< D λ5≤1000 nm;
wherein D TS1< D TD1; D TD1> D TS2; D TS2< D TD2; D TD2> D TS3; and
wherein 380 nm≤ B λ1≤400 nm; 410 nm≤ B λ2≤420 nm;
425 nm≤ B λ3≤435 nm; 465 nm≤ B λ4≤485 nm; and
850 nm≤ B λ5≤1000 nm.

2. The filter system as claimed in claim 1 , wherein:
425 nm≤ D λ1≤435 nm; 465 nm≤ D λ2≤485 nm;
580 nm≤ D λ3≤620 nm; 650 nm≤ D λ4≤770 nm; and
850 nm≤ D λ5≤1000 nm.

3. The filter system as claimed in claim 1 , further comprising:
the first stopband ( D S1) of the detection filter at least partly including the first passband ( B D1) of the illumination filter;
the first passband ( D D1) of the detection filter at least partly including the second passband ( B D2) of the illumination filter; or
the first stopband ( D S1) of the detection filter at least partly including the first passband ( B D1) of the illumination filter and the first passband ( D D1) of the detection filter at least partly including the second passband ( B D2) of the illumination filter.

4. The filter system as claimed in claim 1 , wherein:
B TS1<0.01;
B TD1>0.5;
B TS2<0.1;
B TD2>0.1; and
B TS3<0.01.

5. The filter system as claimed in claim 4 , further comprising:
the third stopband ( B S3) of the illumination filter having a mean transmittance of at most 0.001 in a range from 600 nm to 750 nm.

6. The filter system as claimed in claim 1 , wherein:
D TS1<0.01;
D TD1>0.5;
D TS2<0.1;
D TD2>0.5; and
D TS3<0.01.

7. The filter system as claimed in claim 1 ,
wherein the mean transmittance of at least one of the passbands ( D D1, D D2) is at least 5 times greater than the mean transmittances of the stopbands ( D S1, D S2, D S3) spectrally adjoining the at least one of the passbands.

8. A fluorescence observation system, comprising:
a light production system, which is configured to produce excitation light for exciting fluorescence and illumination light that differs from the excitation light, and to direct the excitation light and the illumination light onto an object region;
a spatially resolving camera for detecting an image of the object region; and
an optical filter system as claimed in claim 1 ;
wherein the illumination filter of the filter system is arranged in a beam path between the light production system and the object region; and
wherein the detection filter of the filter system is arranged in a beam path between the object region and the camera.

9. The fluorescence observation system as claimed in claim 8 , wherein the light production system comprises:
a plurality of light sources, wherein an intensity of light produced by each of the light sources is independently adjustable.

10. The fluorescence observation system as claimed in claim 8 , wherein the light sources for producing excitation light emit substantially exclusively within the stopbands of the detection filter.

11. The fluorescence observation system as claimed in claim 8 , the light production system for producing excitation light comprising at least one of:
a laser with a central emission wavelength of Λ1 or a narrowband violet LED with a central emission wavelength of Λ2,
wherein:
405 nm−ΔΛ Laser ≤Λ1≤405 nm+ΔΛ Laser , with ΔΛ Laser =5 nm;

410 nm−ΔΛ LED ≤Λ2≤410 nm+ΔΛ LED ,with ΔΛ LED =20 nm; and/or

a laser with a central emission wavelength of Λ3 or a narrowband cyan LED with a central emission wavelength of Λ4,
wherein:
488 nm−ΔΛ Laser ≤Λ3≤488 nm+ΔΛ Laser , with ΔΛ Laser =5 nm;

490 nm−ΔΛ LED ≤Λ4≤490 nm+ΔΛ LED , with ΔΛ LED =20 nm; or

a laser with a central emission wavelength of Λ5 or a narrowband IR LED with a central emission wavelength of Λ6,
wherein:
785 nm−ΔΛ Laser ≤Λ5≤785 nm+ΔΛ Laser , with ΔΛ Laser =5 nm;

800 nm−ΔΛ LED ≤Λ6≤800 nm+ΔΛ LED , with ΔΛ LED =20 nm.

12. The fluorescence observation system as claimed in claim 8 , wherein the light production system for producing illumination light comprises at least one of:
a narrowband blue LED with a central emission wavelength in the range from 450 nm−ΔΛ LED to 450 nm+ΔΛ LED ; with ΔΛ LED =20 nm;
a broadband LED, which emits light in the spectral range from 400 nm to 700 nm; or
a narrowband red LED with a central emission wavelength in the range from 660 nm−ΔΛ LED to 660 nm+ΔΛ LED ; with ΔΛ LED =20 nm.

13. The fluorescence observation system as claimed in claim 8 ,
wherein the camera is configured to record a first image of the object region while the excitation light and the illumination light are directed onto the object region.

14. The fluorescence observation system as claimed in claim 13 ,
wherein the camera is configured to record a second image of the object region while only the excitation light or only the illumination light is directed onto the object region.

15. The fluorescence observation system as claimed in claim 14 , further comprising:
a fifth mode of operation,
wherein, in the fifth mode of operation of the fluorescence observation system, a laser with a central emission wavelength of Λ1, 405 nm−ΔΛ Laser ≤Λ1≤405 nm+ΔΛ Laser , ΔΛ Laser =5 nm, or a narrowband violet LED with a central emission wavelength of Λ2, 410 nm−ΔΛ LED ≤Λ2≤410 nm+ΔΛ LED , ΔΛ LED =20 nm, produces the excitation light, and, additionally or alternatively, a laser with a central emission wavelength of Λ3, 488 nm−ΔΛ Laser ≤Λ3≤488 nm+ΔΛ Laser , ΔΛ Laser =5 nm, or a narrowband cyan LED with a central emission wavelength of Λ4, 490 nm−ΔΛ LED ≤Λ4≤490 nm+ΔΛ LED , ΔΛ LED =20 nm, produces the excitation light, and
a broadband LED which emits light in the spectral range from 400 nm to 700 nm produces the illumination light.

16. The fluorescence observation system as claimed in claim 13 , further comprising:
at least one of a first mode of operation, a second mode of operation, and a third mode of operation;
wherein, in the first mode of operation of the fluorescence observation system, a laser with a central emission wavelength of Λ1, 405 nm−ΔΛ Laser ≤Λ1≤405 nm+ΔΛ Laser , ΔΛ Laser =5 nm, or a narrowband violet LED with a central emission wavelength of Λ2, 410 nm−ΔΛ LED ≤Λ2≤410 nm+ΔΛ LED , ΔΛ LED =20 nm, produces the excitation light, and a narrowband blue LED with a central emission wavelength in the range from 450 nm−ΔΛ LED to 450 nm+ΔΛ L

Assignees

• Zeiss Carl Meditec Ag

Inventors

• Nieten Christoph
• Geissler Enrico
• Wicker Kai
• Regensburger Alois
• Jess Helge
• Guckler Roland
• Hauger Christoph

Classifications

• G01N21/6458

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

• G02B21/16

  adapted for ultraviolet illumination {; Fluorescence microscopes (G02B21/0076 takes precedence)} · CPC title

• G01N2021/6471

  Special filters, filter wheel · CPC title

• G01N2201/06113

  Coherent sources; lasers · CPC title

• G01J3/4406 Primary

  Fluorescence spectrometry · CPC title