WO2002034944A1

WO2002034944A1 - High precision and intellectual biochip arrayer having function of respotting

Info

Publication number: WO2002034944A1
Application number: PCT/KR2000/001200
Authority: WO
Inventors: Heui Jae Pahk; Jung Hwan Kim
Original assignee: Dia Chip Limited
Priority date: 2000-10-23
Filing date: 2000-10-23
Publication date: 2002-05-02
Also published as: AU2001210599A1; JP2004512514A

Abstract

The present invention relates to a bio-chip arrayer. The bio-chip arrayer in accordance with the present invention comprises a substrate retaining stand which is separable from the bio-chip arrayer and has a plurality of substrate retaining grooves, wherein each substrate retaining groove includes a retaining edge, a retaining protuberance and a aligning boss for inserting tightly a bio-chip substrate. By using the bio-chip arrayer in accordance with the present invention, the bio-chip substrate may be arrayed on a same position every time. Also, the bio-chip substrate may be re-arrayed by setting up easily a central position and co-ordinates.

Description

HIGH PRECISION AND INTELLECTUAL BIOCHIP ARRAYER HAVING FUNCTION

OF RESPOTTING

Technical Field

The present invention relates to a high precision and intellectual biochip arrayer

with a respotting function, and more particularly to a biochip arrayer capable of precise

respotting through a substrate, retaining grooves formed thereon for accurately retuning a

biochip substrate to its exact previous position, or an optical sensor for precisely

recognizing a reference position so as to define a central position and a coordinate system.

Also, to achieve higher efficiency and wider application, the present invention

relates to a biochip arrayer having a function of precise respotting, comprising an optical

sensor used for recognizing an identifier attached to a biochip so as to extract information

from said identifier such as the type of biochip spot, the procedure used to deposit the spot,

the arrangement of the spot array, the reference position, the spot size, and the intervals

between spots from a database corresponding to the identifier.

Background Art

A biological chip or a biochip is called a biological array. The concept of a biochip

is commonly understood today. A biochip has a substrate having biological material such

as nucleic acid. A deoxyribonucleic acid (DNA) chip is one example of a well-known biochip. The DNA chip has a substrate with DNA fixed thereon. A protein chip is another

example of a biochip and has a substrate with protein formed thereon.

A biochip operates on the basis of the interaction between a target molecule and a

fixed molecule attached to a substrate. For example, the DNA chip operates by means of a

complementary combination of oligonucleotides fixed to a substrate and the bases of the

DNA existing in the sample. Alternatively, a protein chip operates by means of the

interaction between protein molecules such as those found in the antigen-antibody bond or

the ligand-accepter combination.

Because of the recent improvements in DNA sequencing technology, the gene

arrangements of various living things from bacteria to humans are being mapped and vast

quantities of information about the configuration and function of the human genome will

be understood due to the accomplishments of the Human Genome Project. However, it is

difficult to research the hundreds pieces of genetic information newly discovered each day

by means of conventional research methods since these methods require much time and

efforts, therefore, faster and more precise technology is required for researching numerous

genetic databases at the same time.

Therefore, in order to efficiently research genetic information, a DNA chip has

been developed that combines the conventional biological technology, mechanical

automation, and electronic control technology. The DNA chip means the substrate on

which a plurality of DNA is stored at high-density levels, awaiting retrieval. Conventional biological researching techniques only study a small number of

samples at a time. For example, Southern Blot, Northern Blot, mutation retrieval and DNA

sequencing. However, many genes can be more efficiently and automatically analyzed at

once due to the development of the DNA chip.

The DNA chip has many advantages. Enormous amounts of data can be obtained

through a single experiment. Manipulation of the DNA chip and mechanical automation

can easily be accomplished, so research of DNA chips may replace conventional biological

researching methods. Also, the DNA chip can be widely applied to various fields such as

the analysis of the function of genes, identifying genes responsible for causing diseases

like cancer, gene therapy, the quarantine of animals and plants, the testing of food, the

development of new medicines, the retrieval of mutated genes, the analysis of the

arrangement of bases, the testing of tissues, identifying disease causing microorganisms,

forensic medicine, etc.

There are four types of DNA chips, differentiated by the manufacturing method

used to fix the oligonucleotides onto their substrates. They are as follows:

1) Pin micro-array Type: wherein DNA is implanted at identical positions using a

pin,

2) Ink Jet Type: wherein genes contained in a cartridge are injected onto a

substrate by means of electrical force,

3) Photolithography Type: wherein the DNA is directly composed on a substrate by means of photosensitive chemicals, and

4) Electronic array Type: wherein the DNA representing a minus (-) charge is

attached to a predetermined position on a substrate while the material representing a plus

(+) charge is coated.

Also, a fluorescent reader and an electric signal reader are developed as the

apparatus used to collect data contained on a DNA chip.

As it is described above, though many technologies surrounding DNA chips have

been developed for genetic research, advances in the use of DNA chips for the

configuration of an automated disease diagnosis system have not yet been sufficiently

developed.

The immunity diagnosis method of identifying diseases ranges from the utilization

of the blood corpuscle coagulation reagent to the chemical luminescence immunity

measurement method through the radioactive rays immunity measurement method and the

enzyme immunity measurement method.

According to conventional diagnosis methods, after an antigen is fixed and reacted

with an antibody in a sample (mainly, serum) obtained from an entity, the antigen-antibody

reaction is measured by using a secondary antigen composed of a radioactive indicator, an

enzyme and a fluorescent material. The conventional method, however, can measure only a

few samples at a time as discussed previously in the gene analysis method. Hence, it is

difficult to perform simultaneous diagnosis of various diseases, diagnosis for multiple persons, or diagnosis of various diseases in multiple persons. Also, the conventional

methods require, much cost and time since the analysis procedures thereof are not

automated.

Protein chips comprises protein fixed onto a substrate while DNA chips has DNA

fixed onto a substrate. Also, the bound reaction of a protein chip requires multiple

reactions composed of a reaction between an antigen and a primary antibody and a reaction

between the primary antibody and a secondary antibody. In addition, the

separation/combination of the protein chip and the movement of the protein chip can occur

when each reaction transpires inside a chemical reactor.

In general, it is understood that antigen proteins or peptides have various electrical

properties depending on their type and configuration, and are larger than that of the DNA

spot, while DNA has a minus charge and DNA spots range from 15 to 25 bases to about

500bp. Also, proteins are fixed onto a substrate without interfering with, or deforming, the

structure of antigen. Considering such parameters, the method for fixing a protein onto a

substrate is different from that used for DNA, so it is necessary that each antigen protein be

fixed within an optimum range under fixing conditions optimized to facilitate mass

production, and utilizing a detecting procedure capable of ensuring that various antigen

proteins are fixed onto the same substrate, unlike the procedure employed to fix all the

DNA having similar properties onto the same substrate.

In addition, to precisely accomplish a reaction on the surface of the minute substrate, great precision must be taken in placing the protein chip when using the

immunity analysis method for achieving multiple reactions. Therefore, it is important that

the accurate position is chosen whereon to dot the sample in each reaction step.

The conventional biochip arrayer is disclosed as Genosensor Arrays manufactured

by Genosensor Co. in the U.S.A., Biochip Arrayer manufactured by Parkard Instrument Co.

in the U.S.A., Micromax Microarray made by NEN Co. in the U.S.A., and Q array

manufactured by Genetix Co. in the England.

However, the conventional biochip arrayer may not cope with the

separation/combination of the chip and the movement of the chip such as the protein chip

because the conventional biochip arrayer can applied to the DNA chip only. The

conventional biochip arrayer is rarely used today except for experimental purpose.

Also, the conventional biochip arrayer often does not cope with deformations and

errors on the biochip substrate. That is, the conventional biochip arrayer may not overcome

a minute error or deformation generated during the manufacturing process of the biochip

substrate and will not always dot the sample correctly in each reaction step.

Furthermore, the conventional biochip arrayer may not ensure a precise alignment

of the biochip in the biochip arrayer depending on how the sample is fixed to the biochip at

the manufacturer. A protein chip should be precisely aligned while the DNA chip does not

require such precise alignment.

The conventional biochip arrayer has a substrate retaining stand fixed onto the bed thereof, preventing the substrate retaining stand from being easily maintained or repaired.

Disclosure of the Invention

The present invention is intended to overcome the above-described disadvantages.

Therefore, it is an object of the present invention to provide a biochip arrayer that can be

applied to all kinds of biochips such as a deoxyribonucleic acid chip or a protein chip by

using a multiple spindle transfer system.

It is another object of the present invention to provide a biochip arrayer having a

substrate retaining-groove so as to align a biochip substrate to the same position every

times.

It is still another object of the present invention to provide a biochip arrayer having

an aligning boss formed on the substrate retaining-groove for setting a central position and

a coordinate system for the biochip substrate.

a substrate retaining stand including a plurality of substrate retaining-grooves on which

aligning bosses are formed in order to set central positions and coordinates systems for a

plurality of biochip substrates, respectively.

an optical sensor for recognizing predetermined reference points previously indicated on a

biochip substrate so as to set a central position and a coordinate system for the biochip substrate.

It is still another object of the present invention to provide a biochip arrayer that is

highly efficient and widely applicable due to its the optical sensor because its optical

sensor recognizes an identifier on a biochip substrate to which a reference point has been

previously attached, thereby utilizing many pieces of information such as the types of

samples on the biochip, the procedure and an alignment of a spot array, the manufacturer

and diagnosis dates of the biochip, the position of the reference points, and sizes and

intervals of spots after retrieving the information from a database corresponding to the

identifier.

a substrate retaining stand which can be formed separately, thereby facilitating ease of

maintenance or repair done to the substrate retaining stand.

different spots for samples, thereby facilitating simple diagnosis of samples.

To accomplish the objects of the present invention according to one aspect of the

present invention, there is provided a biochip arrayer comprising a substrate retaining stand

having at least one substrate retaining groove for fixing a biochip substrate inserted into the

substrate retaining groove and a bed including at least one well plate retaining groove for

fixing a well plate, wherein the substrate retaining groove comprises a first side for

supporting the biochip substrate by two points or one line, a second side for supporting the biochip substrate by one point or one line, and an aligning boss for receiving a portion of

the biochip substrate wherein the biochip substrate supported by the two points means the

biochip' s substrate is supported by two retaining protuberances that corresponded to the

two points and are formed at predetermined positions on the first side and wherein the

biochip substrate supported by the one point means the biochips substrate is supported by a

retaining protuberance corresponding to the one point and is formed at a predetermined

position on the second side

According to one preferred embodiment of the present invention, the first side

crosses substantially at a right angle or a predetermined angle, with the second side and the

aligning boss is formed between the first side and the second side.

According to another preferred embodiment of the present invention, the substrate

retaining groove further comprises push portions respectively formed on a third side and a

fourth side corresponding to the first side and the fourth side, respectively. At such time,

the push portions respectively support the biochip substrate so that the biochip substrate is

inserted and fixed into the substrate-retaining groove. The push portions are springs or

elastic members, respectively, or a plurality of springs or elastic members, respectively.

In still another preferred embodiment of the present invention, the aligning boss has

a central position for indicating a coordinate point on the biochip substrate. In this case, the

central position is indicated by aligning a home position setting pin, a pin head and the

aligning boss. According to still another preferred embodiment of the present invention, the

substrate-retaining stand is combined separately with the bed. The bed comprises a

washing portion for washing a sample remaining on a probe or the container receiving the

sample.

To accomplish the objects of the present invention according to another aspect of

the present invention, there is provided a method for dotting a biochip wherein a sample is

dotted onto a biochip substrate on a biochip array wherein the first sample has the same

size as the first spot and has been previously dotted so as to be formed onto the biochip

substrate using a biochip arrayer from a manufacturer, which comprises the steps of

disposing the probe mounting part of a biochip arrayer that has been used by a user for

dotting a second sample onto the first spot of the biochip substrate by using the biochip

arrayer of the user, and dotting the second sample onto a second spot corresponding to the

second sample by using the biochip arrayer of the user, wherein the second spot has a

different size than the first spot.

According to one preferred embodiment of the present invention, the step of

dotting the second sample onto the second spot is performed by adjusting the size of a

second probe corresponding to the second spot to differ from the size of the first probe that

corresponds to the first spot, and then dotting the second sample.

According to another preferred embodiment of the present invention, the second

spot is smaller than the first spot when the first spot is an antigen and the second spot is an antibody.

To accomplish the objects of the present invention according to still another aspect

of the present invention, there is provided a method for manufacturing a biochip by using a

biochip array comprising the step of recognizing a biochip identifier indicated on a biochip

substrate, extracting biochip information corresponding to the biochip identifier from a

built-in database wherein the biochip information includes at least one selected from the

group consisting of the total number of spots, the alignment of the spots, the contents of the

spots, the sizes of the spots, the intervals between the spots, the position of a reference spot

and the operating procedure of the biochip arrayer, and dotting the contents of the spots

onto the biochip substrate by using the biochip information.

According to on preferred embodiment of the present invention, the biochip

identifier is recognized from a bar code attached to the biochip substrate. In this case, the

bar code is a first dimension bar code, a second dimension bar code or a third dimension

bar code.

of the present invention, there is provided a biochip arrayer comprising an optical sensor

for recognizing at least one reference point previously indicated on a biochip substrate, a

memory storage and retrieval a program, and a processor combined with the memory to

execute the program, wherein the processor executes in accordance with the program the

step of setting a central position and a coordinate system for the biochip substrate to be retrieved by recognizing at least one reference point and setting coordinates of spots to be

retrieved by using the set central position and the coordinate system.

According to the preferred embodiment of the present invention, the reference

point comprises a first reference value corresponding to the central position, a second

reference value corresponding to an x-axis and a third reference value corresponding a y-

axis.

In another preferred embodiment of the present invention, the optical sensor is an

image detecting element or a position detecting element.

step of setting a central position and a coordinate system on the biochip substrate to be read

by the recognition of at least one reference point, the recognition of positions of spots by

using the set central position, the coordinate system and coordinates of the spots wherein

the coordinates of the spots are previously known, and controlling the dotting of sample in

correspondence with the recognized positions of the spots. Brief Description of the Drawings

The above objects and other advantages of the present invention will become more

apparent by detailed description of the preferred embodiments thereof with reference to the

attached drawings in which:

FIG. 1 is a perspective view showing a biochip arrayer according to one preferred

embodiment of the present invention;

FIG. 2 is a schematic view illustrating a configuration of a control system by using

an optical sensor in the biochip arrayer according to one preferred embodiment of the

present invention;

FIG. 3 is an enlarged side view showing a portion of the biochip arrayer according

to one preferred embodiment of the present invention;

FIG. 4 is an exploded perspective view showing the bed in FIG. 3 according to one

preferred embodiment of the present invention;

FIG. 5 is a perspective view showing a substrate-retaining stand according to one

preferred embodiment of the present invention;

FIGS. 6A is a plane view illustrating a substrate-retaining groove according to one

preferred embodiment of the present invention;

FIG. 6B is a plane view illustrating a substrate-retaining groove according to

another preferred embodiment of the present invention;

FIG. 6C is a plane view illustrating a substrate-retaining groove according to still another preferred embodiment of the present invention;

FIG. 6D is a plane view illustrating a substrate-retaining groove according to still

another preferred embodiment of the present invention;

FIG. 6E is a side view illustrating a spring functioning as a push member in FIGS.

6B to 6D according to one preferred embodiment of the present invention;

FIG. 6F is a side view illustrating an elastic member functioning as a push member

in FIGS. 6B to 6D according to another preferred embodiment of the present invention;

FIG. 7 is a perspective view illustrating a method for setting a portion of an align

boss of a substrate retaining groove to a central position according to one preferred

embodiment of the present invention;

FIG. 8 is a perspective view illustrating a method for aligning a biochip substrate

by using an optical sensor according to one preferred embodiment of the present invention;

FIG. 9 is a plane view for showing a biochip on which a reference point is

indicated according to one preferred embodiment of the present invention;

FIG. 10A is a flow chart illustrating a method for respotting a biochip arrayer by

using an optical sensor according to one preferred embodiment of the present invention;

FIG. 10B is a schematic plane view illustrating a reference representation according

to one preferred embodiment of the present invention;

FIG. 11 is a flow chart illustrating a method for manufacturing a biochip with high

precision and intellectual biochip arrayer recognizing a biochip identifier according to one preferred embodiment of the present invention;

FIG. 12 is a schematic view illustrating a database retrieving biochip information

corresponding to a biochip identifier according to one preferred embodiment of the present

invention;

FIG. 13 is a plane view illustrating a method for the calibration of a bed according

to one preferred embodiment of the present invention;

FIG. 14A is a cross-sectional view illustrating a method for adjusting the height of

a third supporting point according to one preferred embodiment of the present invention;

FIG. 14B is a cross-sectional view illustrating a method for adjusting heights and

y-axis directions of a first and a second supporting points according to one preferred

embodiment of the present invention; and

FIG. 15 is a plane view illustrating a method for adjusting the y-axis direction of a

bed according to one preferred embodiment of the present invention.

Best Modes for carrying out the Invention

Hereinafter, preferred embodiments of the present invention will be described in

more detail with reference to the accompanying drawings, but it is understood that the

present invention should not be limited to the following embodiments.

In the preferred embodiment of the present invention, a biochip is representative of

various biological chips including a deoxyribonucleic acid chip, a protein chip, a ribonucleic acid chip and so on. However, the protein chip will be described as the biochip

hereinafter.

The manufacturing and utilizing processes of the biochip will be described as

follows.

First, a primary protein chip is formed by dotting an antigen onto a biochip arrayer

for a manufacturer. Then, the primary protein chip is dried in a reaction apparatus

maintained at a predetermined temperature and humidity level. In this case, each spot on

the primary protein chip can be identical or different. For example, a portion of the primary

protein chip can hold a human immunodeficiency virus (HIV) antigen, another portion of

the primary protein chip can hold a hepatitis B virus (HBV) antigen and the other portion

the primary protein chip can hold a hepatitis C virus (HCV) antigen.

The primary protein chip is provided to a consumer such as a hospital. The

consumer dots an antigen such as the blood or body fluids of a patient to be tested onto the

primary protein chip to form a secondary protein chip in the biochip arrayer of the

consumer. At that time, the spot of a previously manufactured protein chip should be

placed at an identical position to the spot of the biochip in the arrayer of the consumer.

That is, it is important that the antigen is accurately dotted to the corresponding spot of the

primary protein chip that is next to be realigned. Then, the secondary protein chip is

reacted in the reaction apparatus.

Subsequently, fluorescenin isothiocyanate, a fluorescent material, is dotted onto the secondary protein chip after the user installs the reacted secondary protein chip into the

biochip arrayer of the consumer. In this case, the realignment is required to facilitate

precise respotting.

Then, the result of the reaction can be extracted using a biochip reader, thereby

accomplishing the diagnosis.

embodiment of the present invention.

Referring to FIG. 1, the biochip arrayer comprises a transfer part 103 including a

probe mounting portion 103 whereon at least one probe is mounted, a transfer shaft 101 for

supporting the transfer part 103, a bed 105 and a central control part (not shown) for

controlling those elements. The central control part will be explained with reference to FIG.

2. In this case, the transfer shaft 101 and the transfer part 103 can execute a multiple

spindle transfer under the control of the central control part. Also, the probe and the probe-

mounting portion 105 will be described.

A substrate-retaining stand 109 having a plurality of substrate retaining grooves 111

for fixing several biochip substrates formed onto the bed 107. A washing portion 117 for

washing the probe or a container for receiving samples is formed on the bed 107 and a

washing member 115 having a vacuum suction portion 119 and an ultrasonic washer 121 is

formed on the bed 107. Also, at least one well plate-retaining groove 113 having at least

one well plate formed onto the bed 107. The central control part, having a controller (not shown), is in charge of a relative ^'

position control that determines the relative positions between the probe mounting portion

105, the substrate-retaining groove 111, the well plate retaining groove 113 and the

washing member through a multiple spindle transfer system. At that time, the multiple

spindle transfer system can be applied to all various values of the relative position control

as well as to the case wherein the bed 107 is fixed.

FIG. 2 is a schematic view illustrating a configuration of a control system using an

optical sensor in the biochip arrayer according to one preferred embodiment of the present

invention.

Referring to FIG. 2, the central control part 201 receives a biochip identifier from

an optical sensor and uses the identifier to recognize a chip 207 and receives a reference

point position from an optical sensor, the optical sensor used for recognizing a reference

point 209, and then the central control part 201 sends control signals corresponding to the

biochip identifier and the reference point position to an x-axis transfer driver 213, a y-axis

transfer driver 217, a z-axis transfer driver 221, an ultrasonic driver 223, a vacuum valve

driver 225 and a pump driver 227, respectively. The optical sensor used for recognizing a

chip 207 can be composed of one optical sensor and the optical sensor for recognizing a

reference point 209 can also be composed of one optical sensor. The optical sensor for

recognizing a chip 207 recognizes the biochip identifier from a bar code attached to the

biochip. The central control part 201 includes a standard personal computer 203 and the

controller 205. The controller 205 generates the control signals after the controller 205

receives a communication command and other optical sensor information from the

personal computer 203. Also, the controller 205 receives state information about each

element and transfers the state information to the personal computer 203 or an upper

controller (not shown).

The x-axis transfer driver 213, the y-axis transfer driver 217 and the z-axis transfer

driver 221 respectively control an x-axis motor 211, a y-axis motor 215 and a z-axis motor

219 so as to execute the position control after the x-axis transfer driver 213, the y-axis

transfer driver 217, the z-axis transfer driver 221 receive the control signal from the central

control part 201.

Also, the ultrasonic driver 223 performs washing by generating ultrasonic waves in

the ultrasonic washer 121 after the ultrasonic driver 223 receives the control signal from

the central control part 201. Furthermore, the vacuum valve driver 225 performs washing

by means of a vacuum produced in the vacuum suction portion 119 and the pump driver

227 executes washing by pumping water into the solution washing portion 117.

to one preferred embodiment of the present invention.

As shown in FIG. 3, the biochip arrayer comprises a bed 107, a plurality of probes

301 and a probe-mounting member 105. The probe-mounting member 105 can rotate and move to the right or left. The

probe-mounting member 105 can be established as a general robot arm. In this case, the

probe-mounting member 105 should be bound so that the probe 301 only moves in an

upward or downward motion along the z-axis.

The probe 301 is shaped like a pin having thin and long dimensions so as to spot a

sample onto a biochip substrate. The sample is attached to an end of the probe 301 to be

spotted onto the biochip substrate.

When the biochip substrate is that of a protein chip, the biochip substrate is a solid

plate whose spots are fixed with a number of proteins that are affixed in order. Preferably,

the surface of the biochip is coated.

The solid plate can be composed of glass, deformed silicon, polymer or gel such as

tetraflouroethylene, polystyrene or polypropylene. Preferably, the surface of the substrate

can be coated with a polymer, plastic resin, carbohydrate, silica, silica derivative, carbon,

metal or glass.

The biochip substrate not only supports the samples but also provides the space

wherein the reaction between the fixed sample and a target sample occurs. The dimensions,

the size and the shape of the biochip substrate can be varied in accordance with the purpose

of the analysis and devices employed such as a liquid treating device and a reading wand.

Also, the position of the biochip substrate whereon the samples are fixed can be varied

according to the purpose of the analysis and the types of devices employed. The bed 107 has the biochip substrate affixed thereon and the samples are spotted

onto the biochip. The bed 107 will described in detail with reference to FIG. 4.

preferred embodiment of the present invention.

Referring to FIG. 4, the bed 107 includes the substrate retaining stand 109 having a

plurality of substrate retaining grooves 111, the container washing portion 117, the washing

member 115 (see FIG. 1) having the vacuum suction portion 119 and the ultrasonic washer

121, and the well plate retaining groove 113 whereon at least one well plate is fixed. The

substrate of the substrate retaining groove 111 and the substrate retaining stand 109 means

the biochip substrate.

The substrate-retaining stand 109 is separable from the bed 107. The substrate-

retaining stand 109 can be tightly inserted into a groove formed on a portion of the bed 107

when the groove is formed to receive a portion of the substrate-retaining stand 109. Also,

the substrate-retaining stand 109 can be combined with the bed 107 by means of a screw

combination. Furthermore, the substrate-retaining stand 109 can be combined with the bed

107 by means of a magnet combination.

Since the substrate-retaining stand 109 is combined to, but separately from the bed

107, the substrate retaining stand 109 alone can be separated and changed without it being

necessary to also change the bed 107 when the substrate retaining stand 109 is damaged.

Because of this future, the substrate-retaining stand 109 can be easily maintained. The number of biochip substrates to be mounted can be easily varied because the

substrate-retaining stand 109 can be separated from the bed 107 and the number of the

substrate-retaining groove 111 can be varied. For example, the number of the biochip

substrates can be changed from four to eight when the substrate-retaining stand 109 having

four substrate-retaining grooves 111 is exchanged with the substrate-retaining stand 109

having eight substrate-retaining grooves 111.

The substrate-retaining groove 111 will be explained with reference to FIGS. 6A to

6F.

The washing member 115 (see FIG. 1) washes the samples remaining on the probe

and the container receiving the samples after the samples are spotted onto the biochip. The

container washing portion 117 washes the remaining samples and the container with water.

The vacuum suction portion 119 washes the remaining samples and the container using the

vacuum suction. The ultrasonic washer 121 washes the remaining samples and the

container by using the ultrasonic wave.

The well plate-retaining groove 113 retrieves the samples fixed onto the biochip

where the probe of the probe-mounting portion 105 is inserted into the substrate-retaining

groove 111 to be fixed.

preferred embodiment of the present invention.

As shown in FIG. 5, four substrate-retaining grooves 111 are preferably formed on the substrate-retaining stand 109. The substrate-retaining grooves 111 can be adjusted to

accommodate the size of the biochip substrate.

preferred embodiment of the present invention.

Referring to FIG. 6A, the substrate-retaining groove 111 includes a retaining edge

603, a retaining protuberance 607 and an aligning boss 605.

When a biochip substrate 601 is inserted into the substrate-retaining groove 111, the

biochip substrate 601 is supported at one point by the retaining protuberance 607 and is

supported along one edge by the retaining edge 603.

To fix all biochip substrates 601 at the same position having the same shape, the

retaining edge 603 and the retaining protuberance 607 are formed on the substrate retaining

groove 111 since biochip substrates 601 do not always have the same dimensions and

shape after the biochip substrates 601 are manufactured. It is preferable that one portion of

the substrate-retainmg groove 111 corresponding to the retaining edge 603 is substantially

at a right angle with the other portion of the substrate-retaining groove 111 corresponding

to the retaining protuberance 607.

The aligning boss 605 has the shape of a circle so as to receive a corner of the

biochip substrate 601. Also, the aligning boss 605 performs the function of alignment.

In another preferred embodiment of the present invention, the aligning boss 605

can have various shapes such as an ellipse, a triangle or a tetrahedron, thereby enabling the corner of the biochip substrate 601 to be received.

FIG. 6B is a plane view illustrating a substrate-retaining groove according to

another preferred embodiment of the present invention.

Though the substrate-retainmg groove 111 is substantially identical to that in FIG.

6A, push members 609 are formed on the sides of the retaining edge 603 and the retaining

protuberance 607.

The push members 609 tightly hold the biochip substrate 601 when the biochip

substrate 601 is inserted into the substrate-retaining groove 111. At that time, force is

demanded to tightly insert the biochip substrate 601.

The push members 609 are formed on the substrate-retaining groove 111, but the

push members 609 can be formed in such a way to enable them be separated from the

substrate retaining groove 111 according to another preferred embodiment of the present

invention.

FIG. 6C is a plane view illustrating a substrate-retaining groove according to still

another preferred embodiment of the present invention.

Referring to FIG. 6C, though the substrate-retaining groove 111 is similar to that in

FIG. 6B, the substrate-retaining groove 111 includes two retaining protuberances 607

instead of the retaining edge 603 to support the biochip substrate 601 along one line as

does the retaining edge 603. Two edges of the two retaining protuberances 607 together

support the biochip substrate 601 along one line though the edges of the two retaining protuberance 607 are respectively support the biochip substrate 601 at one point.

FIG. 6D is a plane view for illustrating a substrate-retaining groove according to

still another preferred embodiment of the present invention.

As shown in FIG. 6D, though the substrate -retaining groove 111 is similar to that in

FIG. 6B, the retaining edge 603 is formed instead of the retaining protuberance 607. Hence,

the substrate-retaining groove 111 includes two retaining edges 603 respectively

supporting the biochip substrate 601 along one line.

FIG. 6E is a side view illustrating a spring as a push member in FIGS. 6B to 6D

according to one preferred embodiment of the present invention.

Referring to FIG. 6E, the spring 611 tension presses the biochip substrate 601

firmly to the retaining edge 603 and the retaining protuberance 607 with a predetermined

force.

The push member 609 can be composed of a plurality of springs 611. However,

the force pushing the biochip substrate 601 from one side of the retaining edge 603 should

be divided into two components so as to prevent the biochip substrate 601 from rotating at

the retaining edge 603. Also, the force pushing the biochip substrate 601 from one side of

the retaining protuberance 607 should be directed towards the retaining protuberance 607.

FIG. 6F is a side view illustrating an elastic member as a push member in FIGS.

6B to 6D according to another preferred embodiment of the present invention.

Referring to FIG. 6F, the elastic member 613 generates a predetermined elastic force in one direction along the retaining edge 603 and the retaining protuberance 607 so

that the biochip substrate 601 is tightly held to the substrate-retaining groove 111 when the

biochip substrate 601 is inserted into the substrate-retaining groove 111.

The push member can be composed of a plurality of elastic members 613 similar

in function to the plurality of the springs 611. Also, the force pushing the biochip substrate

601 from one side of the retaining edge 603 should be divided into two components so as

to prevent the biochip substrate 601 from rotating at the retaining edge 603. That is, the

force should be provided without generating momentum at one end of the biochip substrate

601. Also, the force pushing the biochip substrate 601 from one side of the retaining

protuberance 607 should be directed toward the retaining protuberance 607.

The push member 609 is also fixed to the biochip substrate 601 at the same

position and with the same shape since the biochip substrate 601 does not always have the

same dimension and the same shape after the biochip substrate 601 is manufactured.

Referring to FIG. 5 again, a plurality of substrate retaining grooves 111 are formed

on the substrate retaining stand 109 and a plurality of biochip substrates 601 corresponding

to the substrate retaining grooves 111 are inserted into the substrate retaining grooves 111.

In this case, when a portion of one substrate retaining groove 111 from among the

substrate retaining grooves 111 is set as a central position and the positions of the other

substrate retaining grooves 111 are relatively set to the basis of the central position, the

position of the biochip substrate 601 may be difficult to discriminate due to transfer error or thermal deformation error.

Therefore, according to the present invention, a portion of the aligning boss 605

between the substrate-retaining grooves 111 is specifically set to be the central position so

that the positions of the biochip substrates 101 can be determined respectively as positions

to the central position, thereby minimizing the effect due to transfer error and thermal

deformation error. Namely, all the aligning bosses 605 of the substrate retaining grooves

111 retain respectively their central positions so that different coordinate systems are set

respectively for each biochip substrates 601.

The method for setting portions of the aligning bosses 605 of substrate-retaining

grooves 111 to central positions will be described with reference to FIG. 7.

FIG. 7 is a perspective view for illustrating a method for setting a portion of an

aligning boss of a substrate-retaining groove to a central position according to one

preferred embodiment of the present invention.

Referring to FIG. 7, after the pin head 703 is transferred so that the home position

setting pin 701 attachable to the probe mounting member 105 (see FIG. 1) is inserted into a

circular groove of the aligning boss 605 of the substrate-retaining groove 111 and into a

circular groove of the pin head 703, the coordinate is thereby set as the home position of

the biochip substrate 701.

When the above-described work is repeated for each biochip substrate 601,

different coordinate systems can be set for all the biochip substrates 601, thereby minimizing the affect of transfer errors of the apparatus and thermal deformation errors on

the biochip substrate 601.

The biochip array dots the samples by using the central position and the coordinate

system set respectively for each biochip substrate 601.

by using an optical sensor according to one preferred embodiment of the present invention.

As shown in FIG. 8, the optical sensor 801 is attached to the probe mounting

member 105 (see FIG. 1) to detect the reference point previously indicated on the biochip

substrate 805, thereby setting the central position and the coordinate system of the biochip

substrate 805.

In one preferred embodiment of the present invention, the optical sensor 801 can

be an image detecting element or a position detecting element. Preferably, the optical

sensor 801 can be a charged coupled device (CCD) camera. Also, the reference point

previously indicated on the biochip substrate 805 may be represented by all kinds of marks

that the optical sensor 801 recognizes. For example, the reference point can be any marks

printed on the biochip substrate 805, which the CCD camera can recognize.

The biochip array can dot the samples by using the established central position and

the established coordinate system.

FIG. 9 is a plane view showing a biochip on which a reference point is indicated

according to one preferred embodiment of the present invention. Referring to FIG. 9, the reference points 803, 805, and 807 are recognized by the

optical sensor 801, especially the image detecting element, is indicated at a predetermined

position of the biochip substrate 805. In the preferred embodiment of the present invention,

three reference points 803, 804, and 805 are indicated. The central reference point 803 is

recognized as the central position and other reference points 804 and 805 can be set as the

x-axis direction and the y-axis direction. The method for setting the x-axis direction and

the y-axis direction and for setting each position of each spot will be described. The

reference point can be indicated by means of a sample dotted onto the biochip substrate.

For, example, the spot which functions as a reference spot can be set separately from the

setting of other spots.

According to the method described with reference to FIGS. 8 and 9, when the

reference point is indicated on the biochip substrate 805, the biochip arrayer can reset the

central position and the coordinate system for the wrongly positioned biochip substrate 805

by using the optical sensor 801 though the biochip substrate 805 is wrongly positioned on

the bed 107 (see FIG. 1), thereby always dotting the samples onto the correct

predetermined positions on the biochip substrate 805.

The central control part 201 (see FIG. 2) installed in the biochip arrayer for setting

the central position and the coordinate system will be described.

using an optical sensor according to one preferred embodiment of the present invention and FIG. 10B is a schematic plane view illustrating a reference point according to one

preferred embodiment of the present invention.

As shown in FIG. 10A and FIG. 10B, the reference point previously indicated on

the biochip substrate is recognized by means of the optical sensor (step 1001). The

indication of the reference point can be accomplished by the method described in FIG. 10B.

Of course, other indications of the reference point are possible for indicating the central

position and the coordinate system settings.

Reference positions Rl 803, R2 807 and R3 805 corresponding to the reference

point are calculated (step 1003).

A home position coordinate, an x-axis direction vector and a y-axis direction vector

are calculated by using the reference positions Rl 803, R2 807 and R3 805 (step 1005).

The x-axis coordinate of the home position Ox and the y-axis coordinate of the

home position Oy are respectively obtained according to the following equations (1) and

(2).

Ox=Rl.x (1)

Oy=Rl.y (2)

The x-axis direction unit vector Ex and the y-axis direction unit vector Ey are

calculated according to the following equations (3) and (4).

R3 -R1 Ex = (3)

|R3 -R1| R2- R1

Ey = (4)

|R2 - R1| With the calculated home position coordinate, the x-axis direction unit vector Ex

and the y-axis direction unit vector Ey, the real coordinates Px and Py are calculated on the

array of the spot position X and Y according to the following equations (5) and (6).

Px = Ox + X*Ex (5)

Py = Oy + Y*Ey (6)

Preferably, the reference point and the spot position X, Y can be configured into the

biochip arrayer for the manufacturer and the biochip arrayer for the user as previously

promised. That is, the pattern of the spot can be configured into the biochip arrayer for the

manufacturer and the biochip arrayer for the user as previously promised.

In another embodiment of the present invention, the pattern of the spot can be

preset in the biochip arrayer for the manufacturer and recognized in the biochip arrayer for

the user without manually configuring the pattern of the spot in the biochip arrayer for the

manufacturer and the biochip arrayer for the user as previously promised.

In still another embodiment of the present invention, each pattern of each spot can

be set in the biochip arrayer for the manufacturer, and then recognized in the biochip

arrayer for the user.

Then, each motor is controlled in the step 1011 after each numerical control code is generated for controlling each motor in the step 1009.

A precise respot is possible on the biochip substrate if the above-mentioned steps

are implemented.

FIG. 11 is a flow chart illustrating a method for manufacturing a high precision

biochip with a high precision and intellectual biochip arrayer by recognizing a biochip

identifier according to one preferred embodiment of the present invention. The biochip can

be manufactured by means of the controlling the central control part 201.

At first, the biochip identifier indicated on a portion of the biochip substrate is

recognized (step 1101). Preferably, the biochip identifier can be composed of bar codes.

The bar codes can include a one-dimensional bar code, a two-dimensional bar code or a

three-dimensional bar code.

In the step 1103, the biochip information corresponding to the biochip identifier

recognized in the step 1101 is extracted from the built-in database.

The database retrieving the biochip information will be described with reference to

FIG. 12.

invention.

As shown in FIG. 12, the database retrieves various pieces of information such as

the identifier 1201, the total number of spots 1203, the arrangement of the spots 1205, the contents of the spots 1207, the sizes of the spots 1209, the intervals between the spots 1211,

the position of the reference spot 1213 and the operation procedure of the biochip arrayer

1215.

It will be described as follows: the biochip identifier 1201 recognized from the bar

code attached to the biochip substrate is '1'.

When the biochip identifier 1201 is recognized as '1', the biochip information

corresponding to '1' includes the total number of spots 1203 as 100, the arrangement of the

spots 1205 as HOR/ZIG, the content of the spot 1207 as HTBC, the size of the spot 1209

as 500μm, the interval of the spots 1211 as lOOOμm, the position of the reference spot 1213

as (0, 0), (0, 10), (10, 0) and the operation procedure of the biochip arrayer 1215 as SUVSS.

In the arrangement of the spots 1205, HOR/ZIG means the configuration of spots in

zigzags along the horizontal axis and VER/SEQ represents the configuration of the spots in

order along the vertical axis.

In the contents of the spots 1207, HTBC represents HIV/HANTA/Hepaditis B/

Hepaditis C.

As for the operation procedure of the biochip arrayer 1215, S represents the

spotting, U represents the ultrasonic washing and V represents the vacuum suction.

FIG. 13 is a plane view illustrating a method for calibrating a bed according to one

preferred embodiment of the present invention.

Referring to FIG. 13, the first supporting point 1303 and the second supporting point 1305 of the bed 107 adjust the height of the bed 107 and the y-axis direction. The

third supporting point 1307 only adjusts the height of the bed 107. A screw adjusting part

in FIGS. 14A and 14B supports each supporting point, and each supporting point is

indicated as a point.

a third supporting point according to one preferred embodiment of the present invention.

Referring to FIG. 14A, the third supporting point 1307 adjusts the height of the bed

107 by adjusting the screw adjusting part 1403. The screw adjusting part 1403 can

precisely adjust the inclined block with a screw.

FIG. 14B is a cross-sectional view illustrating a method for adjusting the heights

and y-axis direction of a first and a second supporting points according to one preferred

embodiment of the present invention.

Referring to FIG. 14B, the height of the bed 107 is adjusted at the first and the

second supporting points 1303 and 1305 by adjusting the first screw adjusting part 1403. In

addition, the y-axis direction of the bed 107 is adjusted at the first and the second

supporting points 1303 and 1305 by adjusting the second screw adjusting part 1405. The

first screw adjusting part 1403 precisely adjusts the inclined block with a screw. The y-axis

direction of the bed 107 is adjusted by turning the end of the second screw adjusting part

1405 that has a round shape and the bed 107 contacts with the second screw adjusting part

1405 by means of a point contact. The bed 107 is precisely calibrated according to the above-described method with

reference to FIGS. 13, 14A and 14B.

bed according to one preferred embodiment of the present invention.

As shown in FIG. 15, the bed 107 is adjusted along the y-axis direction by

identifying the y-coordinates the coordinates of the aligning bosses in the case that the

coordinates of the aligning bosses of the first and the second substrate-retaining grooves

1501 and 1503 are (125, 25), (25, 25) when the number of the substrate-retaining grooves

is two. Of course, the y-coordinates of the aligning bosses of the substrate retaining

grooves are identified to adjust the bed in the y-axis direction when several substrate-

retaining grooves exist.

To make a precise diagnosis in the biochip arrayer for the user, the diameter of the

first spot dotted onto the biochip substrate in the biochip arrayer for the manufacturer

should preferably different from the diameter of the second spot dotted onto the biochip

substrate in the biochip arrayer for the user. The diameter of the first spot can be different

from that of the second spot because the probe 103 (see FIG. 1) of the biochip arrayer for

the manufacturer a different size than the size of the probe of the biochip arrayer for the

user.

More preferably, the diameter of the first spot is larger than the diameter of the

second spot. Hence, in the biochip reader including the CCD camera for detecting the result of reactions that have occurred in the biochip substrate, the fluorescenin

isothiocynate (FITC) for the CCD camera used to recognize the result of the reaction has a

lower viscosity so that the deficiently in the FITC viscosity can be complemented. At that

time, the first and the second spots respectively become the antigen and the antibody and

the FITC is coated on the resultant antigen-antibody react, when the biochip substrate is

the protein chip.

When the sample is dotted onto a biochip substrate, the biochip substrate is than

the biochip.

While this invention has been described and shown as having multiple designs, the

present invention may be further modified within the spirit and scope of this disclosure.

This application is therefore intended to cover any variations, uses, or adaptations of the

invention using its general principles. Further, this application is intended to cover such

departures from the present disclosure as come within known or customary practice in the

art to which this invention pertains.

Industrial Applicability

According to the present invention, the biochip arrayer has a substrate-retaining

groove enabling the biochip substrate to be aligned at the same position every time.

According to the present invention, the biochip arrayer can cope with the

separation/combination of a protein chip and the movement of a protein chip. According to the present invention, the biochip arrayer has an aligning boss

formed on the substrate-retaining groove for setting the central position and the coordinate

system of the biochip substrate.

stand including a plurality of substrate retaining grooves on which aligning bosses are

formed in order to set the central positions and coordinates systems of a plurality of

biochip substrates, respectively.

According to the present invention, the biochip arrayer has an optical sensor for

recognizing predetermined reference points previously indicated on the biochip substrate

so as to set the central position and the coordinate system of the biochip substrate.

According to the present invention, the biochip arrayer has the substrate-retaining

stand which can be formed separately facilitating ease of maintenance and repair to the

substrate retaining stand.

According to the present invention, the biochip arrayer has different spots for

samples, thereby facilitating the simple diagnosis of samples.

According to the present invention, the biochip arrayer has the multiple spindle

transfer system, thereby making the present invention applicable compatible with all types

of biochips.

Claims

1. A biochip arrayer comprising:

a substrate-retaining stand having at least one substrate-retaining groove for

holding a biochip substrate inserted into said substrate-retaining groove; and

a bed including at least one well plate retaining groove for fixing a well plate,

wherein said substrate-retaining groove comprises:

a first side for supporting said biochip substrate by two points or one line;

a second side for supporting said biochip substrate by one point or one line; and

an aligning boss for receiving a portion of said biochip substrate,

wherein said biochip substrate supported by the two points means said biochip's

substrate is supported by two retaining protuberances that correspond to the two points and

is formed at predetermined positions on said first side and wherein said biochip substrate is

supported by the one points means said biochips substrate is supported by a retaining

protuberance corresponded to the one point and formed at a predetermined position on said

second side

2. The biochip arrayer as claimed in claim 1, wherein said first side

substantially crosses at a right angle or a predetermined angle with said second side.

3. The biochip arrayer as claimed in claim 1, wherein said aligning boss is

formed between said first side and said second side.

4. The biochip arrayer as claimed in claim 1, wherein said substrate retaining

groove further comprises push portions respectively formed on a third side and a fourth

side corresponding to said first side and said fourth side, respectively.

5. The biochip arrayer as claimed in claim 4, wherein said push portions

respectively support said biochip substrate so that said biochip substrate is inserted and

held in said substrate-retaining groove.

6. The biochip arrayer as claimed in claim 4, wherein said push portions are

springs or elastic members, respectively.

7. The biochip arrayer as claimed in claim 6, wherein said push portions are

composed of a plurality of springs or elastic members, respectively.

8. The biochip arrayer as claimed in claim 1, wherein said aligning boss has

a central position for indicating a coordinate point on said biochip substrate.

9. The biochip arrayer as claimed in claim 8, wherein said central position is

indicated by aligning a home position setting pin, a pin head and said aligning boss.

10. The biochip arrayer as claimed in claim 1, wherein said substrate retaining

stand is combined separately with said bed.

11. The biochip arrayer as claimed in claim 1, wherein said bed further

comprises a washing portion for washing a sample remaining on a probe or a container

receiving the sample.

12. A method for dotting a biochip wherein a sample is dotted onto a biochip

substrate on a biochip array in this case a first sample is the same size as a first spot and is

previously dotted onto the biochip substrate using a biochip arrayer for a manufacturer,

which comprises the steps of:

disposing a probe mounting part of a biochip arrayer for a user for dotting a

second sample onto the first spot of said biochip substrate by using said biochip arrayer for

the user; and

dotting the second sample onto a second spot corresponding to the second sample

by using said biochip arrayer for the user, the second spot having a different size from the

size of the first spot.

13. The method for dotting a biochip as claimed in claim 12, wherein the step

of dotting the second sample onto the second spot is performed by adjusting a size of a

second probe corresponding to the second spot to differ from a size of a first probe

corresponding to the first spot so as to dot the second sample.

14. The method for dotting a biochip as claimed in claim 12, wherein the

second spot is smaller than the first spot when the first spot is an antigen and the second

spot is an antibody.

15. A method for manufacturing a biochip by using a biochip array

comprising the steps of:

recognizing a biochip identifier indicated on a biochip substrate;

extracting biochip information corresponding to the biochip identifier from a built-

in database wherein the biochip information includes at least one selected from the group

consisting of total number of spots, alignment of the spots, contents of the spots, sizes of

the spots, intervals between the spots, position of a reference spot and operating procedure

of the biochip arrayer; and

dotting the contents of the spots onto the biochip substrate by using the biochip

information.

16. The method for manufacturing the biochip as claimed in claim 15,

wherein the biochip identifier is recognized from a bar code attached to the biochip

substrate.

17. The method for manufacturing the biochip as claimed in claim 16,

wherein the bar code is a first dimension bar code, a second dimension bar code or a third

dimension bar code.

18. A biochip arrayer comprising:

an optical sensor for recognizing at least one reference point previously indicated

on a biochip substrate;

a memory retrieving a program; and

a processor combined with said memory to execute the program,

wherein said processor executes in accordance with the program the step of setting

a central position and a coordinate system of the biochip substrate to be retrieved by

recognizing at least one reference point, and setting coordinates of spots to be retrieved by

using the set central position and the coordinate system.

19. The biochip arrayer as claimed in claim 18, wherein the reference point comprises a first reference point corresponding to the central position, a second reference

point corresponding to an x-axis and a third reference point corresponding a y-axis.

20. The biochip arrayer as claimed in claim 18, wherein said optical sensor is

an image detecting element or a position detecting element.

21. A biochip arrayer comprising:

on a biochip substrate;

a memory retrieving a program; and

a processor combined with said memory to execute the program,

wherein said processor executes in accordance with the program the step of :

setting a central position and a coordinate system of the biochip substrate to be

retrieved by recognizing at least one reference point;

recognizing positions of spots by using the set central position, the coordinate

system and coordinates of the spots wherein the coordinates of the spots are previously

known; and

controlling dotting of samples in correspondence with the recognized positions of

the spots.

22. The biochip arrayer as claimed in claim 21, wherein the reference point

comprises a first reference point corresponding to the central position, a second reference

23. The biochip arrayer as claimed in claim 21, wherein said optical sensor is

an image detecting element or a position detecting element.

AMENDED CLAIMS

[received by the International Bureau on 8 February 2002 (08.02.02); original claim 15 amended; remaining claims unchanged (1 page)]

13. The method for dotting a biochip as claimed in claim 12, wherein the step

corresponding to the first spot so as to dot the second sample.

14. The method for dotting a biochip as claimed in claim 12, wherein the

spot is an antibody.

15. A method for manufacturing a biochip by using a biochip arrayer

comprising the steps of:

recognizing a biochip identifier indicated on a biochip substrate;

of the biochip arrayer; and

information.