JP4923066B2 - 免疫測定装置、免疫測定装置に用いられる光ファイバ及び免疫測定を行う方法 - Google Patents
免疫測定装置、免疫測定装置に用いられる光ファイバ及び免疫測定を行う方法 Download PDFInfo
- Publication number
- JP4923066B2 JP4923066B2 JP2008558236A JP2008558236A JP4923066B2 JP 4923066 B2 JP4923066 B2 JP 4923066B2 JP 2008558236 A JP2008558236 A JP 2008558236A JP 2008558236 A JP2008558236 A JP 2008558236A JP 4923066 B2 JP4923066 B2 JP 4923066B2
- Authority
- JP
- Japan
- Prior art keywords
- optical fiber
- cavity
- microfluidic channel
- fluorophore
- immunoassay device
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/505—Containers for the purpose of retaining a material to be analysed, e.g. test tubes flexible containers not provided for above
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/05—Flow-through cuvettes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/12—Specific details about manufacturing devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0627—Sensor or part of a sensor is integrated
- B01L2300/0654—Lenses; Optical fibres
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/087—Multiple sequential chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N2021/0346—Capillary cells; Microcells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
- G01N2021/6467—Axial flow and illumination
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6484—Optical fibres
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Hematology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- Dispersion Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Description
Claims (25)
- 少なくとも1つのマイクロ流体チャンネルを有する免疫測定装置において用いられる光ファイバであって、
励起光を前記マイクロ流体チャンネルへ伝送するとともに、発せられた蛍光を光検出器へ伝送するための光ファイバであり、
当該光ファイバは、
(a)少なくとも1つのキャビティであって、各キャビティが検体を保持するとともに反応チャンバとしての機能を果たすキャビティを備え、
前記キャビティは、前記マイクロ流体チャンネル内へ開放しており該マイクロ流体チャンネルの一部を形成し、前記キャビティは、当該光ファイバのシースを貫いて当該光ファイバのコアの中に設けられた溝である、光ファイバ。 - 検体流の方向と平行な方向で光を光検出器に伝送する請求項1に記載の光ファイバ。
- 前記少なくとも1つのマイクロ流体チャンネルの壁の少なくとも一部を形成する請求項1および2のいずれか一項に記載の光ファイバ。
- 当該光ファイバ内を前記キャビティに伝送される励起光が前記キャビティ内のフルオロフォアを励起できる請求項1に記載の光ファイバ。
- 前記フルオロフォアによって発せられる蛍光が前記キャビティから当該光ファイバの出射端に当該光ファイバ内を伝送される請求項4に記載の光ファイバ。
- 前記フルオロフォアがフルオレセインである請求項4または5に記載の光ファイバ。
- 前記キャビティが前記少なくとも1つのマイクロ流体チャンネル内にある状態で、当該光ファイバが前記少なくとも1つのマイクロ流体チャンネル内へ突出する請求項1〜6のいずれか一項に記載の光ファイバ。
- 前記少なくとも1つのマイクロ流体チャンネルのベースの少なくとも一部を形成する請求項1〜7のいずれか一項に記載の光ファイバ。
- 当該光ファイバ内で伝送される励起光が、前記フルオロフォアの励起波長に基づいて選択される波長を有している請求項4〜8のいずれか一項に記載の光ファイバ。
- 蛍光を使用して免疫測定を行なう方法であって、
a)少なくとも1つのマイクロ流体チャンネルを有する免疫測定装置内に光ファイバを設けるステップであって、前記光ファイバが、少なくとも1つのキャビティを備え、各キャビティが検体を保持するとともに反応チャンバとして機能し、前記キャビティは、前記マイクロ流体チャンネル内へ開放しており該マイクロ流体チャンネルの一部を形成し、前記キャビティは、前記光ファイバのシースを貫いて前記光ファイバのコアの中に設けられた溝である、ステップと、
b)前記少なくとも1つのマイクロ流体チャンネルに沿って複数の検体を通過させるステップであって、前記複数の検体のうちの少なくとも1つがフルオロフォアを含むステップと、
c)前記少なくとも1つのキャビティ内で前記複数の検体を反応させるステップと、
d)前記フルオロフォアが蛍光を発するように前記キャビティ内の前記フルオロフォアを励起するために、前記少なくとも1つのキャビティに前記光ファイバ内の励起光を伝送するステップと、
e)前記発せられた蛍光を光ファイバに沿って前記キャビティから前記光ファイバの出射端に伝送するステップと、
f)前記蛍光を検出するステップと、
を備える方法。 - 前記励起光が、前記発せられた蛍光を残すように前記検出でフィルタリングされる請求項10に記載の方法。
- 前記励起光および前記発せられた蛍光が、前記少なくとも1つのマイクロ流体チャンネル内での検体流の方向と略平行な方向で伝送される請求項10または11に記載の方法。
- 前記光ファイバが前記少なくとも1つのマイクロ流体チャンネルの壁の少なくとも一部を形成する請求項10〜12のいずれか一項に記載の方法。
- 前記フルオロフォアがフルオレセインである請求項10〜13のいずれか一項に記載の方法。
- 前記励起光の波長が前記フルオロフォアの励起波長に基づいて選択される請求項10〜14のいずれか一項に記載の方法。
- 少なくとも1つのマイクロ流体チャンネルと、少なくとも1つの光ファイバとを有する免疫測定装置であって、
前記光ファイバが、励起光を前記マイクロ流体チャンネルへ伝送するとともに、発せられた蛍光を光検出器へ伝送し
前記光ファイバは、
(a)少なくとも1つのキャビティであって、各キャビティが検体を保持するとともに反応チャンバとしての機能を果たすキャビティを備え、
前記キャビティは、前記マイクロ流体チャンネル内へ開放しており該マイクロ流体チャンネルの一部を形成し、前記キャビティが、前記光ファイバのシースを貫いて前記光ファイバのコアの中に設けられた溝である、免疫測定装置。 - 前記光ファイバが検体流の方向と平行な方向で光を光検出器に伝送する請求項16に記載の免疫測定装置。
- 前記光ファイバが前記少なくとも1つのマイクロ流体チャンネルの壁の少なくとも一部を形成する請求項16および17のいずれか一項に記載の免疫測定装置。
- 前記光ファイバ内を前記キャビティに伝送される励起光が前記キャビティ内のフルオロフォアを励起できる請求項16に記載の免疫測定装置。
- 前記フルオロフォアによって発せられる蛍光が前記キャビティから前記光ファイバの出射端に前記光ファイバ内を伝送される請求項19に記載の免疫測定装置。
- 前記フルオロフォアがフルオレセインである請求項19または20に記載の免疫測定装置。
- 前記キャビティが前記少なくとも1つのマイクロ流体チャンネル内にある状態で、前記光ファイバが前記少なくとも1つのマイクロ流体チャンネル内へ突出する請求項16〜21のいずれか一項に記載の免疫測定装置。
- 前記光ファイバが前記少なくとも1つのマイクロ流体チャンネルのベースの少なくとも一部を形成する請求項16〜22のいずれか一項に記載の免疫測定装置。
- 前記光ファイバ内で伝送される励起光が、前記フルオロフォアの励起波長に基づいて選択される波長を有している請求項19〜23のいずれか一項に記載の免疫測定装置。
- 請求項1〜9のいずれか一項に記載の光ファイバを備える免疫測定装置。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/SG2006/000044 WO2007102783A1 (en) | 2006-03-07 | 2006-03-07 | Microfluidic immunoassay device |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2009529137A JP2009529137A (ja) | 2009-08-13 |
JP4923066B2 true JP4923066B2 (ja) | 2012-04-25 |
Family
ID=38475148
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008558236A Expired - Fee Related JP4923066B2 (ja) | 2006-03-07 | 2006-03-07 | 免疫測定装置、免疫測定装置に用いられる光ファイバ及び免疫測定を行う方法 |
Country Status (5)
Country | Link |
---|---|
US (1) | US8158363B2 (ja) |
EP (1) | EP2002240A1 (ja) |
JP (1) | JP4923066B2 (ja) |
AU (1) | AU2006339611A1 (ja) |
WO (1) | WO2007102783A1 (ja) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2009335000B2 (en) | 2008-12-30 | 2014-11-13 | Children's Medical Center Corporation | Method of predicting acute appendicitis |
KR101519379B1 (ko) * | 2010-04-29 | 2015-05-12 | 삼성전자 주식회사 | 원심력 기반의 미세유동장치 및 이를 이용한 면역분석방법 |
WO2012058716A1 (en) * | 2010-11-02 | 2012-05-10 | Edith Cowan University | An optical sensor for measuring a property of a fluid |
US8958071B2 (en) | 2011-11-15 | 2015-02-17 | Empire Technology Development Llc | Integrated optical sensor |
GB201120769D0 (en) | 2011-12-02 | 2012-01-11 | Biochrom Ltd | Improvements in and relating to devices for recieving liquid samples |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4844869A (en) * | 1985-09-09 | 1989-07-04 | Ord, Inc. | Immunoassay apparatus |
JPS62123358A (ja) * | 1985-11-22 | 1987-06-04 | Sumitomo Electric Ind Ltd | 光フアイバ型免疫センサ |
JP3130513B2 (ja) * | 1989-04-19 | 2001-01-31 | イビデン株式会社 | 生体活性物質測定用装置 |
US5340715A (en) * | 1991-06-07 | 1994-08-23 | Ciba Corning Diagnostics Corp. | Multiple surface evanescent wave sensor with a reference |
EP0991777A1 (en) * | 1997-06-18 | 2000-04-12 | Ulrich J. Krull | Nucleic acid biosensor diagnostics |
EE04249B1 (et) * | 1999-04-21 | 2004-02-16 | Asper O� | Meetod biopolümeermaatriksi lugemiseks ja fluorestsentsdetektor |
WO2001051207A1 (en) * | 2000-01-10 | 2001-07-19 | Genospectra, Inc. | Linear probe carrier |
US7175811B2 (en) * | 2000-04-28 | 2007-02-13 | Edgelight Biosciences | Micro-array evanescent wave fluorescence detection device |
FR2809491B1 (fr) * | 2000-05-26 | 2008-07-04 | Production Rech S Appliquees | Procede et appareil de metrologie ellipsometrique pour echantillon contenu dans une chambre ou analogue |
CA2314398A1 (en) * | 2000-08-10 | 2002-02-10 | Edward Shipwash | Microarrays and microsystems for amino acid analysis and protein sequencing |
US6600558B2 (en) | 2000-08-22 | 2003-07-29 | Nippon Telegraph And Telephone Corporation | Micro-fluidic cell for optical detection of gases and method for producing same |
US20020072111A1 (en) * | 2000-12-13 | 2002-06-13 | Clarkin James P. | Drawn microchannel array devices and method of analysis using same |
US6949176B2 (en) | 2001-02-28 | 2005-09-27 | Lightwave Microsystems Corporation | Microfluidic control using dielectric pumping |
US6974673B2 (en) * | 2001-09-24 | 2005-12-13 | Veridian Systems Division | Coupled capillary fiber based waveguide biosensor |
US7350569B2 (en) * | 2004-06-14 | 2008-04-01 | Weatherford/Lamb, Inc. | Separable plug for use in a wellbore |
TWI251079B (en) * | 2004-06-15 | 2006-03-11 | Univ Nat Cheng Kung | Inspection chip and the manufacturing method thereof |
US7799278B2 (en) * | 2004-07-06 | 2010-09-21 | Schlumberger Technology Corporation | Microfluidic system for chemical analysis |
-
2006
- 2006-03-07 AU AU2006339611A patent/AU2006339611A1/en not_active Abandoned
- 2006-03-07 US US12/282,166 patent/US8158363B2/en not_active Expired - Fee Related
- 2006-03-07 EP EP06717168A patent/EP2002240A1/en not_active Withdrawn
- 2006-03-07 JP JP2008558236A patent/JP4923066B2/ja not_active Expired - Fee Related
- 2006-03-07 WO PCT/SG2006/000044 patent/WO2007102783A1/en active Application Filing
Also Published As
Publication number | Publication date |
---|---|
JP2009529137A (ja) | 2009-08-13 |
AU2006339611A1 (en) | 2007-09-13 |
US20090194707A1 (en) | 2009-08-06 |
US8158363B2 (en) | 2012-04-17 |
EP2002240A1 (en) | 2008-12-17 |
WO2007102783A1 (en) | 2007-09-13 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US6137117A (en) | Integrating multi-waveguide sensor | |
US9989466B2 (en) | Method for reducing analyzer variability using a normalization target | |
CN101057134A (zh) | 用于生物检测的系统和方法及其微共振传感器 | |
RU2446402C2 (ru) | Обнаружение целевых молекул в образце | |
JP4923066B2 (ja) | 免疫測定装置、免疫測定装置に用いられる光ファイバ及び免疫測定を行う方法 | |
US20080179540A1 (en) | Surface plasmon enhanced fluorescence sensor | |
US8964183B2 (en) | Systems and methods for screening of biological samples | |
KR101957800B1 (ko) | 정량적 측정을 위한 스트립 삽입형 형광 리더기 | |
US10871447B2 (en) | Bleaching of dyes in luminescent detection | |
US10436707B2 (en) | Detection of analytes using nanoparticles as light scattering enhancers | |
EP3705875B1 (en) | An apparatus and method for detecting photoluminescent light emitted from a sample | |
US10845310B2 (en) | Quantification of topologically arranged luminescent dyes | |
JP4480130B2 (ja) | 光学分析装置 | |
EP3317650B1 (en) | Bleaching of dyes in luminescent detection | |
US10928305B2 (en) | Modulation of luminescent dyes | |
EP3317643B1 (en) | Modulation of luminescent dyes | |
KR20200040417A (ko) | 정량적 측정을 위한 스트립 삽입형 형광 리더장치 및 장치의 프로세서 | |
Aksoy et al. | Emerging Technologies for Fluorescence-Based Optical Test Strip Readers | |
JP2024002206A (ja) | 物質分離装置、分析装置及び分析方法 | |
CN116829925A (zh) | 光学模块 | |
JP2009150665A (ja) | 一分子蛍光・りん光分析方法及びそのための装置 | |
Fluorosensors et al. | Instrumentation for Cylindrical Waveguide |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110927 |
|
A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20111226 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20120124 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20120206 |
|
R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150210 Year of fee payment: 3 |
|
LAPS | Cancellation because of no payment of annual fees |