TWI633183B - Bioparticle capture chipset - Google Patents

Abstract

The present invention provides a bioparticle capture wafer set for capturing target biological particles. The bioparticle capture wafer set comprises a pair of transparent carriers having a microchannel, and a spoiler microstructure located in the microchannel, and the interspersed microstructures on the two transparent carriers are allowed to flow through the microchannel The liquid sample produces a spoiler effect and flows uniformly through the microchannel and is captured in the microchannel. In addition, since the wafer assembly body is made of a transparent material, the target biological particles captured in the micro flow channel can be subjected to omnidirectional observation analysis directly from the micro flow channel.

Description

Bioparticle capture chipset

The present invention relates to a particle capture wafer, and more particularly to a biological particle capture wafer.

The current fetal chromosome bioassay in the mother can be roughly classified into invasive detection and non-invasive detection. For example, amniocentesis is an invasive examination of fetal chromosome detection in the mother. It is taken from the amniotic fluid surrounding the developing fetus because the amniotic fluid contains the skin of the fetus and other sample cells that are shed during development. The test can be performed on the chromosomes in these cells. Non-invasive examinations typically involve the extraction of a small amount of maternal peripheral blood for fetal chromosome detection. Since the invasive detection method is a complete sample taken directly from the fetus, the accuracy is unquestionable, but the invasive detection still has a risk of abortion of 0.1 to 0.2%. The biggest advantage of the non-invasive examination is safety, but still Problems with accuracy need to be overcome.

For the aforementioned non-invasive examination, the most active promotion in the market is Non-Invasive Prenatal Diagnosis (NIPD). NIPD is free fetal DNA (cffDNA, cell-free fetal) DNA) is a screening sample. cffDNA is a small part of the incomplete DNA fragment (~60-200 bp) formed by apoptosis of placental cells, which is then released in the maternal blood. Since cffDNA is a DNA fragment, it is necessary to use mathematical algorithms to try to piece together the complete DNA sequence of the fetus, in which the concentration of cffDNA is a key factor. When the concentration is too low, there is a possibility of detecting failure or false-negative. The aforementioned NIPD, the current study also pointed out that there are fetal cells in the peripheral blood of pregnant women, fetal nucleated red blood cells (fNRBCs) or placental trophoblast cells (trophoblast) as fetal chromosome examination, can simultaneously It has two major advantages: safety and high correctness.

In the current study, the method of enrichment of fetal cells such as nucleated red blood cells is mainly based on flow cytometry. However, most of these nucleated red blood cells are maternal, and the fe birth is low. Density gradient separations can significantly increase fetal cell yield. It is composed of non-transparent or translucent materials for related research and capture technologies, so it is not conducive to observing and analyzing the captured cells.

Accordingly, it is an object of the present invention to provide a bioparticle capture wafer set for capturing target biological particles and for omnidirectional observation of the captured target biological particles.

Thus, the bioparticle capture wafer set of the present invention comprises a wafer body, The wafer body defines at least one microchannel having a relatively distant inlet and an outlet, and has a spoiler microstructure located in the at least one microchannel, and the wafer body is made of a transparent material.

The effect of the invention is that the spoiler microstructure of the microchannel is used to generate a spoiler effect on the sample fluid flowing through the microchannel, and uniformly flows through the microchannel and is captured in the spoiler microstructure. . In addition, the wafer body is further made transparent, so that the target biological particles captured in the microchannel can be observed in all directions directly from the microchannel.

2‧‧‧chip body

21‧‧‧microchannel

241‧‧‧Second board body

242‧‧‧Second spoiler block

211‧‧‧ entrance

212‧‧‧Export

22‧‧‧Spoiler microstructure

23‧‧‧First carrier

231‧‧‧ first board body

232‧‧‧First spoiler block

24‧‧‧second carrier

25‧‧‧ joints

26‧‧‧ rough microstructure

27‧‧‧ Ligand layer

100‧‧‧ board

101‧‧‧ transparent layer

D1, d2‧‧‧ distance

Other features and advantages of the present invention will be apparent from the embodiments of the present invention. FIG. 1 is a schematic plan view showing an embodiment of the biological particle capturing wafer set of the present invention; FIG. 2 is a secant line of FIG. FIG. 3 is a schematic view showing the manufacturing process of the first and second spoiler blocks of the embodiment; FIG. 5 is a schematic view showing another embodiment of the first and second spoiler blocks of the embodiment; Further included is a schematic diagram of a roughened microstructure; FIG. 6 illustrates a schematic diagram of the embodiment further comprising a ligand layer.

Before the present invention is described in detail, it should be noted that in the following description, similar elements are denoted by the same reference numerals.

The bioparticle capture wafer set of the present invention is a target biological particle that can be used to capture a liquid sample. The liquid sample may be a body fluid derived from an animal, such as blood, lymph, saliva, urine, etc., but is not limited thereto. The target biological particle may be a specific cell, such as fetal nucleated red blood cells (fNRBC), fetal trophoblast cells, circulating tumor cells (CTC), viruses or bacteria, etc. There are no special restrictions.

Referring to Figures 1 and 2, an embodiment of the biological particle capture wafer set of the present invention comprises a wafer body 2. 2 is a partial cross-sectional view of the a-a secant line of FIG. 1. The wafer body 2 is made of a transparent material, and defines a plurality of microchannels 21, and each of the microchannels 21 has an inlet 211 and an outlet 212 that are relatively far apart, and has a turbulent flow located in the microchannel 21. Structure 22. It should be noted that the micro flow channel 21 of the wafer body 2 may also be a single strip. When the micro flow channel 21 is plural, the micro flow channels 21 may be independent of each other or may be connected to each other. In this embodiment, the wafer body 2 defines a plurality of microchannels 21 that can communicate with each other as an example.

In detail, the wafer body 2 has a first carrier 23, a second carrier 24, and a joint portion 25 for connecting the first and second carrier plates 23, 24.

The first carrier 23 has a first board body 231 and a plurality of first spoilers 232 protruding from the first board body 231 and transmitting light. The second carrier 24 has a second board body. 241 and a plurality of second spoilers 242 protruding from the second plate body 241 and transmitting light. The first and second plate bodies 231, 241 are mutually connected to each other with the surfaces of the first and second spoiler blocks 232, 242 to mutually define the micro flow channels 21, and the first and second spoiler blocks 232, 242 are respectively located in the microchannels 21, and the perturbation microstructures 22 are collectively formed in each of the microchannels 21.

The first and second plate bodies 231, 241 may be made of a transparent polymer material such as polymethyl methacrylate (PMMA), polymethyl siloxane (PDMS), or polycarbonate (PC), or It is made of a transparent material such as glass. The first and second spoiler blocks 232, 242 are selected from transparent materials different from the first and second plate bodies 231, 241, such as transparent photoresist (SU8), tetraethoxy decane (Tetra Ethyl-Ortho) -Silicate, TEOS), or spin-on-glass (SOG), etc., and the first and second spoiler blocks 232, 242 can be square, trapezoidal, or triangular. Or semicircular. The arrangement of the first and second spoiler blocks 232, 242 allows a liquid sample flowing through the microchannel 21 to generate a spoiler effect, thereby allowing the liquid sample to flow more uniformly through the microfluid. Road 21. 1 and 2 are diagrams in which the structures of the first and second spoiler blocks 232 and 242 are trapezoidal, but the actual implementation is not limited to this shape.

Referring to FIG. 3, the manufacturing method of the foregoing embodiment is preceded by two pieces. The surface of the plate body 100 is formed of a transparent material such as a transparent photoresist (SU8), a Tetra Ethyl-Ortho-Silicate (TEOS), or a spin-on-glass (SOG). After the transparent layer 101 is further combined with the characteristics of the transparent material, the predetermined portion of the transparent layer 100 is removed by lithography or etching, and then formed on the two transparent plates 100 respectively. The first and second spoiler blocks 232, 242 are shown, and the joint portions 25 located outside the first and second spoiler blocks 232, 242, respectively, to form the first and second carrier plates 23, 24. Then, the first and second plate bodies 231 and 241 are relatively bonded to each other by the bonding portion 25 by using a transparent adhesive, thereby completing the production of the biological particle capturing chip set.

Referring to FIG. 2, preferably, the distance d1 of the adjacent two sides of any adjacent first and second spoiler blocks 232, 242 is twice the diameter of the target biological particles, and the first and second spoiler blocks 232, 242 The distance d2 to the surface of the opposing second and a plate bodies 241, 231 is twice the diameter of the target biological particle. By the control of the equal distances d1 and d2, the first and second spoiler blocks 232 and 242 adjacent to the micro flow channel 21 and the first and second spoiler blocks 232 and 242 can be arranged. A connected capture space is formed between the opposing second and a plate bodies 241, 231, and thus the target biological particles are captured as they flow through the capture space. The target biological particles captured in the microchannel 21 are transparent to the wafer body 2, so that the wafer body 2 can be directly observed through the wafer body 2 without the need to capture the target biological particles as conventionally used. The wafer is opaque or translucent, and it is necessary to observe the target biological particles. The disadvantage of taking the target biological particles out of the wafer.

In addition, referring to FIG. 4, it should be noted that the first and second spoiler blocks 232 and 242 may be formed by using the foregoing methods, or may be separately etched or sandblasted from the two boards 100. The surface of the board 100 is removed from the predetermined portion, and the first and second spoilers 232, 242 and the joints 25 are formed, and the first, as shown in FIG. The second carrier plates 23, 24 are only the same constituent materials of the first and second spoiler blocks 232, 241 formed in this manner.

Preferably, referring to FIG. 5, regardless of the manner in which the first and second spoiler blocks 232, 242 are formed, the embodiment of the present invention is located in the micro flow channel 21, and the first and second disturbances are not formed. The surfaces of the first and second plate bodies 231 and 241 of the flow blocks 232 and 242 may be further roughened by etching, sand blasting or the like to form a roughened microstructure 26. The use of the roughened microstructures 26 further increases the contact area of the target biological particles within the microchannels 21, thereby improving the target bioparticle capture efficiency.

In addition, referring to FIG. 6, the embodiment of the biological particle trapping wafer set of the present invention may further be on the surface of the first and second plate bodies 231, 241 located in the micro flow channel 21, or the first and second surfaces. The surface of the spoiler blocks 232, 242, or at least one of the surfaces of the roughened microstructures 26, is coated with a layer of ligand 27 that can interact with the target biological particles, for example, the ligand layer 27 can be Streptavidin produces a ligand layer that can affinity with the captured target biological particles for more precise capture Capture target biological particles. In FIG. 6, the description is made by taking the ligand layer 27 formed on the roughened microstructures 26.

In summary, the present invention utilizes the microchannel 21 to provide a spoiler microstructure 22 having spoiler and trapping particle characteristics, so that the sample fluid flowing through the microchannel 21 has a spoiler effect and uniformly flows through The microchannel 21 is captured by the spoiler microstructure 22. Further, since the wafer body 2 of the present invention is transparent, the target biological particles trapped in the microchannel 21 can be directly subjected to the microchannel 21. In addition, the present invention can further form a roughened microstructure 26 in the microchannel 21, and further form a ligand layer 27 on the roughened microstructure 26, by the roughening. The microstructure 26 can increase the contact area of the target biological particles in the microchannel 21, and improve the capture efficiency of the target biological particles, and the ligand layer 27 can more accurately capture the target biological particles, so The cost can be achieved for the purpose of the invention.

However, the above is only the embodiment of the present invention, and the scope of the invention is not limited thereto, and all the equivalent equivalent changes and modifications according to the scope of the patent application and the patent specification of the present invention are still The scope of the invention is covered.

Claims (6)

A biological particle capture wafer set for capturing target biological particles, comprising: a wafer body defining at least one microchannel having an inlet and an outlet relatively far away, and having at least one microflow The first slab has a first slab and a second slab. The first slab has a first slab and a plurality of slabs. a first spoiler block that protrudes and transmits light from the first plate body, the second carrier plate has a second plate body and a plurality of second spoiler blocks protruding from the second plate body and transmitting light, The first board body and the second board body are respectively connected to each other with the surfaces of the first spoiler block and the second spoiler block to define the at least one micro flow channel, and the first spoiler block And the second spoiler block is located in the at least one micro flow channel to jointly form the spoiler microstructure, and the first board body and the second board body and the first spoiler block and the second The spoiler block may be composed of the same or different materials; the first plate body and the second The body is located on the surface of the at least one microchannel and further has a roughened microstructure; the biological particle capturing wafer set further has a ligand formed on the surface of the roughened microstructure and bonded to the target biological particle Floor. The bioparticle capture wafer set of claim 1, wherein the first spoiler block and the second spoiler block have a side view structure that is square, trapezoidal, triangular, or semi-circular. The bioparticle capture chip set of claim 1, wherein the first plate body and the second plate body and the first spoiler block and the second spoiler block are made of different materials, and Waiting for the first spoiler block and the second spoiler The material of the block is selected from the group consisting of transparent photoresist, tetraethoxy decane, or spin-on glass. The biological particle capture chip set of claim 1, wherein the distance between the first spoiler block and the second spoiler block to the opposite surface of the second plate body and the first plate body is 2 times the diameter of the target biological particles. The biological particle capturing wafer set according to claim 4, wherein a distance between adjacent ones of the adjacent first spoiler blocks and the second spoiler blocks is twice the diameter of the target biological particles. The bioparticle capture wafer set of claim 1, wherein the wafer body defines a plurality of microchannels having an inlet and an outlet that are relatively distant, and each of the microchannels has a spoiler microstructure.