US 5,062,942 AGranted

Fluorescence detection type electrophoresis apparatus

AssigneeHitachi, Ltd.

InventorsHideki Kambara, Tetsuo Nishikawa, Tomoaki Sumitani, Keiichi Nagai, Yoshitoshi Ito

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Abstract

In a fluorescence detection type electrophoresis apparatus, comprising an electrophoresis separation gel plate (2, 102, 205), an excitation laser beam source (3, 201, 201) for emitting fluorescent light, and a fluorescent light detector (9, 109, 212) for detecting the emitted fluorescent light, a fluorescent light images are divided into a plurality of virtual images by an image divider (5, 7.sub.2, 104, 105, 209) and at the same time, the lights corresponding to the individual divided images are wavelength-selected by bandpass filters (8, 8.sub.3, 107, 108, 120, 227-230), thereby providing highly accurate, sensitive separation and detection of DNA fragments and the like and determination of base sequence of DNA and the like.

Claims25

Dependent claims

A fluorescence detection type electrophoresis apparatus, comprising:

(i) a migration lane on which sample fragments labeled with fluorophore migrate electrophoretically;
(ii) means for irradiating locally a predetermined portion to be irradiated with an excitation light for exciting said fluorophore, said predetermined portion being on said migration lane;
(iii) optical means for focusing a fluorescent light image which is emitted from said fluorophore at said predetermined irradiated portion, on a plurality of focusing positions separated from each other, said optical means comprising:
a) a focusing lens disposed on a predetermined distance from said predetermined irradiated portion;
and
b) an image dividing means disposed on an optical path from said predetermined irradiated portion to said focusing lens and makes said fluorescent light image visible as a plurality of virtual images positionally different from each other when said light image is seen from the focusing lens side;
iv) a plurality of wavelength selection means disposed on the respective optical paths from said predetermined irradiated portion to said plurality of focusing positions, said respective optical paths lying separately from each other at the position of said wavelength selection means, and said plurality of wavelength selection means having transmission wavelength band different from each other;
and
v) a light detector for detecting with positional distinction each light amount of a plurality of fluorescent light images each of which had passed through each of said wavelength selection means and focused on each of said focusing positions.

6 more elements

A fluorescent detection type electrophoresis apparatus comprising:

i) a platelike migration lane on which a plurality of sample fragments migrate electrophoretically, said plurality of sample fragments being labeled respectively with a plurality of fluorophores having different maximum emission wavelengths from each other;
ii) means for irradiating locally a predetermined portion to be irradiated with at least one laser beam for exciting said plurality of fluorophores, said predetermined portion being on said migration lane and having a bandlike shape with a longitudinal side perpendicular to the migration direction of said fragments;
iii) optical means for focusing a bandlike fluorescent light image, which is emitted from the fluorophore at said predetermined irradiated portion, on a plurality of focusing positions separated from each other, said optical means comprising:
a) a focusing lens disposed on a predetermined distance from said predetermined irradiated portion;
and
b) an image dividing means which is disposed on an optical path from said predetermined irradiated portion to said focusing lens and which makes said fluorescent light image visible as a plurality of virtual images positionally different from each other when said light image is seen from the focusing lens side;
iv) a plurality of band-pass filters disposed on the respective optical paths from said predetermined irradiated portion to said plurality of focusing positions, said respective optical paths lying separately from each other at the positions of said band-pass filters, and each of said plurality of band-pass filters selecting a fluorescent light emitted from one of said fluorophores from the other fluorescent lights emitted from the other fluorophores with respect to the wavelength and transmitting the selected fluorescent light;
and
v) a light detector for detecting with positional distinction each light amount of a plurality of band-like fluorescent light images which has passed through each of said band-pass filters and focused on each of said focusing positions, to thereby detect each of said plurality of sample fragments in passing through said predetermined irradiated portion.

6 more elements

A fluorescent detection type electrophoresis apparatus comprising:

i) a platelike migration lane on which sample fragments labeled with fluorophore migrate electrophoretically;
ii) means for irradiating locally a predetermined portion to be irradiated with a least one laser beam for exciting said fluorophores, said predetermined portion being on said migration lane and having a bandlike shape with a longitudinal side perpendicular to the migration direction of said fragments;
iii) a prism having an incidence plane on which the fluorescent light emitted from said fluorophore at the predetermined irradiated portion is made to be incident and having a plurality of light-outgoing planes;
iv) a focusing lens which is disposed at a position capable of viewing said predetermined irradiated portion though said plurality of light-outgoing planes of said prism and by which said fluorescent lights outgoing from said plurality of light out-going planes focus respectively on focusing positions different from each other to thereby form a plurality of respective bandlike image;
v) a plurality of wavelength selection means disposed on the respective optical paths from said predetermined irradiated portion to said plurality of focusing positions, said respective optical paths lying separately from each other at the position of said wavelength selection means, and said plurality of wavelength selection means having transmission wavelength bands different from each other;
and
vi) a light detector for detecting with positional distinction each light amount of a plurality of bandlike fluorescent light images each of which has passed through each of said wavelength selection means and focused on each of said focusing positions.

4 more elements

A fluorescent detection type electrophoresis apparatus comprising:

i) a platelike migration lane on which a plurality of sample fragments migrate electrophoretically, said plurality of sample fragments being labeled respectively with a plurality of fluorophores having different maximum emission wavelengths from each other;
ii) means irradiating locally a predetermined portion to be irradiated with at least one laser beam for exciting said plurality of fluorophores, said predetermined portion being on said migration lane and having a bandlike shape with a longitudinal side perpendicular to the migration direction of said fragments;
iii) a cylindrical convex lens converging a fluorescent light emitted from said fluorophore at said predetermined irradiated portion, in a width direction of said predetermined irradiated portion of bandlike shape;
iv) a prism having an incidence plane on which the fluorescent light transmitted through said cylindrical convex lens is made to be incident and having a plurality of light-outgoing planes;
v) a cylindrical concave lens diverging a fluorescent light outgoing from said plurality of light-outgoing plane of said prism, in a width direction of said predetermined irradiated portion of bandlike shape;
vi) a focusing lens which is disposed at a position capable of viewing said predetermined irradiated portion of bandlike shape through said cylindrical concave lens, said prism and said cylindrical convex lens and which focuses said fluorescent lights outgoing from said plurality of light-outgoing planes respectively on focusing positions different from each other to thereby form respectively a plurality of bandlike images;
vii) a plurality of band-pass filters disposed on the respective optical paths from said predetermined irradiated portion to a plurality of said focusing portions, said respective optical paths lying separately from each other at the positions of said band-pass filters, and each of said plurality of band-pass filters, and each of said plurality of band-pass filters selecting a fluorescent light emitted from one of said fluorophores from the other fluorescent lights emitted from the other fluorophores with respect to the wavelength and transmitting the selected fluorescent light;
and
viii) a light detector for detecting with positional distinction each light amount of a plurality of bandlike fluorescent light images which has passed through each of said band-pass filters and focused on each of said focusing positions, to thereby detect each of said plurality of sample fragments in passing through said predetermined irradiated portion.

6 more elements

Dependencies are inferred from the claim text.