CN102911860A - Portable fluorecyte counting device - Google Patents
Portable fluorecyte counting device Download PDFInfo
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- CN102911860A CN102911860A CN2012103719370A CN201210371937A CN102911860A CN 102911860 A CN102911860 A CN 102911860A CN 2012103719370 A CN2012103719370 A CN 2012103719370A CN 201210371937 A CN201210371937 A CN 201210371937A CN 102911860 A CN102911860 A CN 102911860A
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Abstract
The invention provides a portable fluorecyte counting device, the portable fluorecyte counting device comprises a sampling unit composed of an array micro-fluidic chip and a sample stage, wherein antibodies are marked in the micro channel of the array micro-fluidic chip in order to catch target cells; the sample stage is used for loading the array micro-fluidic chip; a fluorescence detection unit including an LED (light-emitting diode) as laser light source, a laser filter and an emission filter used for filtering light, and a CCD (charge coupled device) used for collecting fluorescent light and transforming the result into picture form; a stepping motor for driving the sample stage to move; a digital signal processing unit used for performing data processing of the picture and controlling signal acquisition of the CCD and operation of the stepping motor; and a power supply. The portable fluorecyte counting device is convenient to carry and has the advantages of quick analysis, high flux and easy operation.
Description
Technical field
The present invention relates to a kind of fluorocyte counting mechanism, specifically a kind of portable high throughput fluorescence cell counter.
Background technology
Cell counting all has very important significance in Basic of Biology research and clinical diagnostic applications.In the yeast culturing process, for its speed of growth is measured, need the quantity of its different steps is monitored; In the drug research, often need the quantitative expedition medicine to the action effect of target cell, need quantitatively cell to be counted; Need to detect the number of all kinds cell in the patient blood for the state of an illness of understanding the common cold people; For learning whether cancer patient exists cancer cells diffusion, also need count cancer cells in its blood in the hospital; Cell counting seems particularly important for the leukaemic, for the quantity of accurately learning the hematopoiesis osteocyte in the patient body in the chemotherapy process at any time treatment plan is revised, the doctor needs repeatedly count the cell in the blood samples of patients.Above all explanations are to various types of cells, and especially the fast and accurately quantitative counting of some specific rare cells seems unusually important, and develop can quick counter and light easy-operating quantitative cell counting instrument seem particularly urgent.
Being used for Cytometric decision method has multiple, such as counting afterwards by dyeing, perhaps cell is carried out fluorescent mark and counted afterwards, perhaps directly count according to cell size, perhaps having or not nucleus to count according to cell, is a kind of very effective method and count based on cell fluorescence.Because it is high that fluoroscopic examination has sensitivity, the advantage that selectivity is good, in many researchs, be employed, such as flowing of some membranins on the cytolemma, the research of intracellular signaling pathway, the compartment analysis of all kinds of materials in the cell, the folding research of protein, analysis of microorganism etc. all can use fluorescence and comes target substance is detected in the water quality.
At present, more existing Cytometric devices.Such as cell counting count board (patent 200920126697.1), carry out people's number number by after dyeing at microscopically, although this method Financial cost is low, analysis speed is slow, flux is low and human cost consumption is large; Also be useful in addition Cytometric flow cytometer (patent 200810038044.8), the fluorescence of this instrument Main Basis target cell judges, analytic function is powerful, but instrument is expensive, and is bulky, needs special messenger's operation.Therefore, existing these cell counters are difficult to the particularly demand of clinical field of satisfied instantly each field at present.
Summary of the invention
The purpose of this invention is to provide a kind of portable high throughput fluorescence cell counter, have carrying portable, analyze fast, flux is high and easy-operating characteristics.
The technical solution adopted in the present invention is: a kind of Portable fluorescence cell counter, it is characterized in that, and comprising:
Give the sample unit, be comprised of array micro-fluidic chip and sample stage, antibody on the mark in its array micro-fluidic chip microchannel is used for the captured target cell; Sample stage is used for carrying microarray micro-fluidic chip;
Fluorescence detection unit comprises the LED as excitation light source, excites filter disc and emission filter disc as what filter, as phosphor collection and the result is converted into the CCD of picture form;
Stepper-motor is used for driving the movement of sample stage;
Digital signal processing unit is used for the data processing of picture and controls the signals collecting of whole CCD and the operation of stepper-motor;
And power supply, for all need the power supply of electric parts in the instrument.
Aforesaid Portable fluorescence cell counter, it is characterized in that, digital signal processing unit is comprised of digital signal processor DSP and associated electronics, be used for the start-up and operation of LED, CCD and stepper-motor is controlled, and the resulting as a result figure of CCD can be converted into readable data.
The invention has the beneficial effects as follows: as Cytometric sample preparation unit, the cell capturing efficiency is high with the array micro-fluidic chip that is marked with antibody in the present invention, and high specificity and sample consumption are few; Pass through transmission shaft Quality control Stage microscope with stepper-motor simultaneously, can realize the movement of array micro-fluidic chip, thereby guarantee high-throughout cell counting; Realize the fluorescent signal collection with CCD, and optical signal is converted into picture, signals collecting and processing speed are fast; Carry out digital signal processing with digital signal processing unit, can realize result's synchronous conversion and the ordered control of whole device.
Description of drawings
Fig. 1 is the schematic appearance of the Portable fluorescence cell counter of the embodiment of the invention.
Fig. 2 is the structural representation of the Portable fluorescence cell counter of the embodiment of the invention.
Fig. 3 is the schematic diagram of the fluorescence detection unit of the embodiment of the invention.
Fig. 4 is the schematic diagram of the display micro-fluidic chip of the embodiment of the invention.
Fig. 5 is the sample detection process schematic diagram of the embodiment of the invention.
Embodiment
In order to understand better the present invention, further illustrate content of the present invention below in conjunction with embodiment, but content of the present invention not only is confined to the following examples.Those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values are equally within the listed claims limited range of the application.
Description of symbols among the figure: 1-to sample unit, 2-fluorescence detection unit, 3-stepper-motor, 4-power supply, 5-digital signal processing unit, 21-excitation light source, 22-excite filter disc, 23-emission filter disc, 24-CCD, 6-array micro-fluidic chip, 7-power supply, 8-case lid display screen, 9-start button, 10-stop button.
Referring to Fig. 2, portable high throughput testing device of the present invention comprises to sample unit 1, fluorescence detection unit 2, stepper-motor 3, power supply 4 and digital signal processing unit 5.
The described sample unit 1 of giving comprises array micro-fluidic chip and sample stage.The microchannel internal labeling of array micro-fluidic chip has antibody, is used for the captured target cell.As required can be with different antibody in its array microchannel internal labeling.After cell suspending liquid added array micro-fluidic chip, be placed on the sample stage.A reaction member in the elder generation pair array microchannel detects, and after having detected, stepper-motor drives sample stage by transmission shaft and moves a distance, allows second reaction member of microchannel be in detected position.
Described fluorescence detection unit 2 is located at the sample stage top, and the LED by as excitation light source 21 excites filter disc 22, and emission filter disc 23 and CCD 24 form.Light that LED sends carries out sample excitation through after exciting filter disc, sends fluorescence behind the sample reception photon, is collected by CCD after fluorescence filters through the emission filter disc and this optical signal is converted into the gray scale picture.
Digital signal processing unit 3 is comprised of digital signal processor (DSP) and associated electronics, is used for the start-up and operation of LED, CCD and stepper-motor is controlled, and the resulting as a result figure of CCD can be converted into readable data.
In order conveniently to use and to carry, add a shell in the complete assembly outside, as shown in Figure 1.The shell top arranges user interface, and the housing right side arranges external power interface, can link to each other from different adaptor power supplies.User interface comprises display interface and operation interface, and operation interface can be touch screen type or button.Plug in, press the startup button on the operation interface, LED begins luminous, CCD claps the first pictures, after first sample unit of microchannel had gathered, CCD stopped to take pictures, and stepper-motor drives the distance that sample stage moves a unit by transmission shaft, allow second sample unit be in detected position, CCD stops to take pictures after clapping the second pictures; Still rotate with stepper-motor afterwards, Stage microscope moves, and stops the detection that this program is finished all samples unit after CCD takes pictures.After all samples unit inspection was complete, stepper-motor drove sample stage and gets back to starting position, and CCD closes, and LED closes.This moment, simultaneous display provided the concentration value of target cell on display screen as a result.If need to detect without next group sample, can press OFF switch, pull out power supply.
From the one-piece construction arrangement, control panel is positioned at the upper left surface of whole body structure, digital signal processing unit is positioned at the position, lower-left of casing, fluorescence detection device is positioned at the inboard on the upper right surface of casing, sample stage is positioned at the centre of casing right half part, and stepper-motor and power supply lay respectively at the place ahead and the rear of sample stage.
The detection principle of fluorocyte proofing unit of the present invention is: the target cell surface markers has fluorescent substance, is fixed on the specific antibody that is adsorbed onto microchannel surface.The photon that LED sends excites fluorescent substance through the emission filter disc, and fluorescent substance absorbs exciting light and radiates the long fluorescence of wavelength, and fluorescence is gathered by CCD and can form a bright spot at the picture that generates, i.e. corresponding target cell.For the sample unit of each microchannel, there are how many target cells to form corresponding bright spot number at picture, after can being identified by digital signal processing unit, the bright spot number on this pictures directly is converted to corresponding numeral; Second sample unit is picked in the same way to be gone out the target cell number and adds up with the numeral of a upper sample unit.Total number of target cell in the sample can be obtained after a plurality of sample units have been detected, the concentration of target cell can be obtained with the volume of this numeral and institute's sample introduction product.
The present invention is described further below by a concrete embodiment.
Example 1: the concentration of HeLa cell (cervical cancer cell) in the test sample.
HeLa cell and HePG2 cell (liver cancer cell) are the cells that is commonly used for experimental study.We will be marked with the HeLa cell of green fluorescence material and not have the HePG2 cell of mark fluorescent to mix with certain proportion.To be example to this sample detect to show specific implementation process of the present invention to the concentration of HeLa cell to the below.
Choosing of each element of light path test section:
Green fluorescence material maximum excitation wavelength in the HeLa cell is about 480 nm, and maximum emission wavelength is about 520 nm.We choose
Light source led: great power LED, centre wavelength are 480 nm;
Exciter filter: peak transmission wavelength 480nm, half-band width 15 nm;
Emission spectral filter: peak transmission wavelength 525nm, half-band width 15 nm;
Operation steps:
I, this example with an array micro-fluidic chip with 4 sample units as example, as shown in Figure 4.4 reaction members are arranged on the microchannel, linearly arrange.Fixed volume (being set to 0.1 milliliter here) sample is injected from feeder connection, and sample can flow in four reaction members along the microchannel.
The case lid on II, opening unit the right as shown in Figure 5, is placed on the sample stage array micro-fluidic chip 6 to regard to the position, then the right case lid is covered.
III, as shown in Figure 5 plugs in 7, presses the start button 9 on the control panel, and this device is started working.
IV, given an order by digital signal processing unit, light LED and send exciting light, picture is collected and be converted into to the fluorescence that target cell in first sample pool sends through CCD, corresponding numeral is picked up and be converted into to the picture processing degree that bright spot on the picture is installed by digital signal processing unit, namely finished the detection of first sample unit, CCD stops to take pictures; At this moment, dependent instruction can allow the stepping machine operation drive the distance that sample stage moves a sample unit by transmission shaft, so that second sample unit is in detected position.CCD restarts to take pictures, and gathers the second pictures and target cell number and is converted into corresponding numeral by digital signal processing unit, and CCD stops to take pictures.The cumulative summation of gained target cell number and previous sample unit numerical value in second sample unit.
V, repeating step VI are finished successively the detection of other sample units and are obtained the total N of target cell.At this moment, device is long-pending according to total cellular score and population of samples, and the concentration that demonstrates target cell at case lid display screen 8 is N/0.1 mL-1.
VI, press and stop button 10, pull out power supply.
The content that is not described in detail in this specification sheets belongs to the known prior art of this area professional and technical personnel.
Claims (2)
1. a Portable fluorescence cell counter is characterized in that, comprising:
Give the sample unit, be comprised of array micro-fluidic chip and sample stage, antibody on the mark in its array micro-fluidic chip microchannel is used for the captured target cell; Sample stage is used for carrying microarray micro-fluidic chip;
Fluorescence detection unit comprises the LED as excitation light source, excites filter disc and emission filter disc as what filter, as phosphor collection and the result is converted into the CCD of picture form;
Stepper-motor is used for driving the movement of sample stage;
Digital signal processing unit is used for the data processing of picture and controls the signals collecting of whole CCD and the operation of stepper-motor;
And power supply, for all need the power supply of electric parts in the instrument.
2. Portable fluorescence cell counter according to claim 1, it is characterized in that, digital signal processing unit is comprised of digital signal processor DSP and associated electronics, be used for the start-up and operation of LED, CCD and stepper-motor is controlled, and the resulting as a result figure of CCD can be converted into readable data.
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| CN2012103719370A CN102911860A (en) | 2012-09-29 | 2012-09-29 | Portable fluorecyte counting device |
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| CN2012103719370A CN102911860A (en) | 2012-09-29 | 2012-09-29 | Portable fluorecyte counting device |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232468A (en) * | 2014-08-28 | 2014-12-24 | 中北大学 | Laser array coded and photoinduced cell separation device |
| CN104263644A (en) * | 2014-08-28 | 2015-01-07 | 中北大学 | Cell separation method based on laser array coding and photo-induction |
| CN104280329A (en) * | 2014-09-01 | 2015-01-14 | 上海柏慧康生物科技有限公司 | Micro-fluidic multicolor fluorescence cell counter |
| CN106248942A (en) * | 2016-08-31 | 2016-12-21 | 隗慧林 | A kind of urine Exfoliated tumor cells Dual channel detection equipment |
| CN108645778A (en) * | 2018-05-24 | 2018-10-12 | 广西民族大学 | A kind of portable CD+4T lymphocytes detecting system and its automated cell counting algorithm |
| CN109030839A (en) * | 2018-05-24 | 2018-12-18 | 广西民族大学 | A kind of light fluid attachment based on smart phone |
| WO2022247945A1 (en) * | 2021-05-28 | 2022-12-01 | 上海睿钰生物科技有限公司 | Counting chamber and application, and method and system for analyzing particles in sample to be tested |
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| CN202830038U (en) * | 2012-09-29 | 2013-03-27 | 武汉斯坦姆赛尔生物技术有限公司 | Portable fluorescence cell counting device |
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Patent Citations (1)
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| CN202830038U (en) * | 2012-09-29 | 2013-03-27 | 武汉斯坦姆赛尔生物技术有限公司 | Portable fluorescence cell counting device |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232468A (en) * | 2014-08-28 | 2014-12-24 | 中北大学 | Laser array coded and photoinduced cell separation device |
| CN104263644A (en) * | 2014-08-28 | 2015-01-07 | 中北大学 | Cell separation method based on laser array coding and photo-induction |
| CN104232468B (en) * | 2014-08-28 | 2016-03-30 | 中北大学 | Laser array coding and photoinduced cell separation apparatus |
| CN104263644B (en) * | 2014-08-28 | 2016-07-06 | 中北大学 | Laser array coding and photoinduced cell isolation method |
| CN104280329A (en) * | 2014-09-01 | 2015-01-14 | 上海柏慧康生物科技有限公司 | Micro-fluidic multicolor fluorescence cell counter |
| CN106248942A (en) * | 2016-08-31 | 2016-12-21 | 隗慧林 | A kind of urine Exfoliated tumor cells Dual channel detection equipment |
| CN108645778A (en) * | 2018-05-24 | 2018-10-12 | 广西民族大学 | A kind of portable CD+4T lymphocytes detecting system and its automated cell counting algorithm |
| CN109030839A (en) * | 2018-05-24 | 2018-12-18 | 广西民族大学 | A kind of light fluid attachment based on smart phone |
| WO2022247945A1 (en) * | 2021-05-28 | 2022-12-01 | 上海睿钰生物科技有限公司 | Counting chamber and application, and method and system for analyzing particles in sample to be tested |
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Application publication date: 20130206 |