ES2330068A1 - DEVICE FOR VISUALIZATION OF FLUORESCENT BIOLOGICAL MATERIAL. - Google Patents
DEVICE FOR VISUALIZATION OF FLUORESCENT BIOLOGICAL MATERIAL. Download PDFInfo
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- ES2330068A1 ES2330068A1 ES200603288A ES200603288A ES2330068A1 ES 2330068 A1 ES2330068 A1 ES 2330068A1 ES 200603288 A ES200603288 A ES 200603288A ES 200603288 A ES200603288 A ES 200603288A ES 2330068 A1 ES2330068 A1 ES 2330068A1
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- 239000012620 biological material Substances 0.000 title claims abstract description 14
- 238000012800 visualization Methods 0.000 title claims abstract description 8
- 230000005284 excitation Effects 0.000 claims abstract description 10
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 claims description 5
- 230000003287 optical effect Effects 0.000 claims description 4
- 238000001228 spectrum Methods 0.000 claims description 4
- 241000195493 Cryptophyta Species 0.000 claims description 3
- 238000001917 fluorescence detection Methods 0.000 claims description 3
- 241000252212 Danio rerio Species 0.000 claims description 2
- 241000244206 Nematoda Species 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 238000004113 cell culture Methods 0.000 claims description 2
- 210000002257 embryonic structure Anatomy 0.000 claims description 2
- 231100001261 hazardous Toxicity 0.000 claims description 2
- 241000269370 Xenopus <genus> Species 0.000 claims 1
- 230000000712 assembly Effects 0.000 claims 1
- 238000000429 assembly Methods 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 11
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 abstract description 2
- 229910052753 mercury Inorganic materials 0.000 abstract description 2
- 229910052724 xenon Inorganic materials 0.000 abstract description 2
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 abstract description 2
- 230000008878 coupling Effects 0.000 abstract 1
- 238000010168 coupling process Methods 0.000 abstract 1
- 238000005859 coupling reaction Methods 0.000 abstract 1
- 239000003086 colorant Substances 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001143 conditioned effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pathology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Heart & Thoracic Surgery (AREA)
- Medical Informatics (AREA)
- Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
Dispositivo para visualización de material biológico fluorescente. En el seno de un armazón o carcasa (1) se establece al menos un diodo emisor de luz (2) y, enfrentado al mismo, un filtro de excitación (3), enfrentados a su vez a un rehundido superior (5) de dicha carcasa para el correcto coplamiento de un porta (6) con la muestra a analizar, mientras que al menos un filtro de emisión (8) se sitúa a un nivel superior, sobre un brazo soporte (7) de configuración en "U", y queda también enfrentado a los elementos anteriores. Se consigue de esta manera, frente a los sistemas convencionales que utilizan lámparas de mercurio o de xenón, una drástica reducción tanto de costes como de volumen, permitiendo además realizar la visualización del material fluorescente en el mismo lugar de trabajo habitual.Device for visualization of fluorescent biological material. Within a frame or casing (1) at least one light-emitting diode (2) is established and, facing it, an excitation filter (3), facing in turn an upper recess (5) of said housing for the correct coupling of a holder (6) with the sample to be analyzed, while at least one emission filter (8) is located at a higher level, on a support arm (7) with a "U" configuration, and it is also faced with the previous elements. In this way, compared to conventional systems that use mercury or xenon lamps, a drastic reduction in both costs and volume is achieved, also allowing fluorescent material to be displayed in the same usual workplace.
Description
Dispositivo para visualización de material biológico fluorescente.Device for material display Biological fluorescent.
La presente invención se refiere a un dispositivo especialmente concebido para la visualización de material biológico fluorescente, aplicable al sector de la investigación en los campos de la biología, la química, la farmacia y la agricultura.The present invention relates to a device specially designed for viewing biological fluorescent material, applicable to the sector of research in the fields of biology, chemistry, pharmacy and agriculture
El dispositivo es aplicable para diversos materiales biológicos, ya sean organismos completos (por ejemplo larvas y adultos del insecto Drosophila, embriones de pez cebra, nematodos, algas, etc), tejidos o cultivos celulares, incluyendo los cultivos de bacterias, levaduras, microalgas, etc... Por tanto se trata de un equipo versátil destinado a su uso en laboratorios de investigación.The device is applicable for various biological materials, whether they are whole organisms (for example larvae and adults of the Drosophila insect, zebrafish embryos, nematodes, algae, etc.), tissues or cell cultures, including cultures of bacteria, yeasts, microalgae, etc ... Therefore it is a versatile equipment intended for use in research laboratories.
El objeto de la invención es conseguir un dispositivo de dimensiones reducidas y bajo costo, tanto de fabricación como de mantenimiento, que permite la visualización de material biológico fluorescente de forma fácil, rápida y continua en el tiempo.The object of the invention is to achieve a Small device and low cost of both manufacturing as maintenance, which allows the display of fluorescent biological material easily, quickly and continuously in the time.
Hasta la fecha, el rastreo de material fluorescente "in situ" se lleva a cabo con fuentes de luz de diferentes tipos que permitieran obtener una iluminación intensa de pequeñas porciones de la muestra a analizar, siendo las más empleadas las lámparas de mercurio o xenón. Dichas lámparas tienen un alto coste, son voluminosas, requieren largos periodos de pre-calentamiento y su vida es muy limitada (entre 200 y 300 horas).To date, the tracking of fluorescent material " in situ " is carried out with light sources of different types that allow to obtain an intense illumination of small portions of the sample to be analyzed, the most commonly used being mercury or xenon lamps. These lamps have a high cost, are bulky, require long periods of pre-heating and their life is very limited (between 200 and 300 hours).
Además, dichas fuentes se encuentran generalmente acopladas a microscopios de fluorescencia o lupas, así como a sistemas de adquisición y análisis de imágenes lo que aumenta sustancialmente el precio del equipamiento total necesario para la función de rastreo de dicho material.In addition, these sources are found usually coupled to fluorescence microscopes or magnifiers, as well as to image acquisition and analysis systems what substantially increases the price of the total equipment needed for the tracking function of said material.
Así pues, los sistemas actuales para visualización de material biológico fluorescente requieren de equipos voluminosos y caros, con poca versatilidad y notables costos de mantenimiento, que además generalmente no permiten realizar la visualización del material fluorescente en el mismo lugar de trabajo habitual.So, the current systems for Visualization of biological fluorescent material require bulky and expensive equipment, with little versatility and remarkable maintenance costs, which also generally do not allow perform the visualization of the fluorescent material in it usual workplace.
El dispositivo que la invención propone resuelve de manera plenamente satisfactoria la problemática anteriormente expuesta, en los diferentes aspectos comentados.The device proposed by the invention resolves in a fully satisfactory way the problem previously exposed, in the different aspects commented.
Para ello y de forma más concreta dicho dispositivo está basado en el uso de diodos emisores de luz (conocidos como LED, de las siglas en inglés de Light Emitting Diode) como fuente de luz, en combinación con filtros de emisión y excitación, lo que trae consigo un dispositivo estructuralmente simple y consecuentemente económico, de reducido tamaño, que permite realizar la visualización del material fluorescente en el mismo lugar de trabajo habitual, con un rastreo del material rápido y sencillo.To do this and more specifically said device is based on the use of light emitting diodes (known as LED, of the acronym in English of Light Emitting Diode) as a light source, in combination with emission filters and excitation, which brings with it a structurally device simple and consequently economical, small in size, which allows to visualize the fluorescent material in the same usual workplace, with fast material tracking And simple.
Adicionalmente, todo el espectro de colores fluorescente puede ser visualizado por medio de los filtros y de los LED adecuados, con adaptación a las exigencias específicas de cada caso.Additionally, the entire spectrum of colors fluorescent can be visualized by means of filters and the right LEDs, adapting to the specific requirements of each case.
La utilización de LED, cuya vida media es de 20.000 horas, permite un uso continuo y rutinario de larga duración, su reducido tamaño permite trabajar en un espacio también reducido, bajo una lupa de disección o dentro de una campana de cultivo, pudiendo además adaptarse o integrarse en equipos previamente existentes, como es el caso de los difusores (o "trampas") de CO_{2} normalmente empleados para anestesiar adultos de la mosca Drosophila (ampliamente utilizado en estudios de genética y desarrollo). Además y mediante la integración de la fuente de luz en un armazón impermeable, es posible el trabajo con especimenes y muestras que contengan sustancias o compuestos peligrosos para la salud.The use of LEDs, whose half-life is 20,000 hours, allows a continuous and routine use of long duration, its small size allows to work in a space also reduced, under a magnifying glass or inside a culture hood, and can also adapt or integrate into previously existing equipment, such as the diffusers (or "traps") of CO2 normally used to anesthetize adults of the Drosophila fly (widely used in genetics and development studies). In addition and by integrating the light source into an impermeable framework, work with specimens and samples containing substances or compounds hazardous to health is possible.
Se ha previsto también la posibilidad de equipar el dispositivo con una cámara situada sobre la pieza soporte de los filtros de emisión.The possibility of equipping is also planned the device with a camera located on the support part of the emission filters
También se ha previsto la posibilidad de añadir un fotodiodo para detección de fluorescencia en la misma pieza soporte en la que se encuentran los filtros de emisión.It is also planned to add a photodiode for fluorescence detection in the same piece support on which the emission filters are located.
Se ha previsto igualmente la posibilidad de insertar una platina para mover muestras situadas en placas multi-pocillo, lo que supone una gran mejora en el rastreo del material múltiple que puede ser acondicionado en dichas placas.The possibility of insert a stage to move samples located on plates multi-well, which is a great improvement in the tracking of multiple material that can be conditioned in said plates.
Para complementar la descripción que se está realizando y con objeto de ayudar a una mejor comprensión de las características del invento, de acuerdo con un ejemplo preferente de realización práctica del mismo, se acompaña como parte integrante de dicha descripción, un juego de dibujos en donde con carácter ilustrativo y no limitativo, se ha representado lo siguiente:To complement the description that is being performing and in order to help a better understanding of the characteristics of the invention, according to a preferred example of practical realization of it, is accompanied as an integral part of said description, a set of drawings where with character Illustrative and not limiting, the following has been represented:
Figura 1.- Muestra, según una vista general en perspectiva, un dispositivo para visualización de material biológico fluorescente realizado de acuerdo con el objeto de la presente invención, concretamente de acuerdo con la variante de realización más simple de la invención.Figure 1.- Sample, according to an overview in perspective, a device for viewing material Biological fluorescent performed according to the purpose of the present invention, specifically in accordance with the variant of simplest embodiment of the invention.
Figura 2.- Muestra una vista en alzado lateral del mismo dispositivo.Figure 2.- Shows a side elevation view of the same device.
Figura 3.- Muestra, una representación esquemática en perfil de los elementos funcionales del dispositivo.Figure 3.- Sample, a representation schematic profile of the functional elements of the device.
Figura 4.- Muestra, según una perspectiva similar a la figura 1, una variante de realización del dispositivo en la que éste incorpora además una trampa de CO_{2}.Figure 4.- Sample, according to a perspective similar to figure 1, a variant embodiment of the device in which it also incorporates a CO2 trap.
Figura 5.- Muestra, también según una vista en perspectiva, el dispositivo de la figura 4 al que se ha añadido además el soporte para una cámara digital.Figure 5.- Sample, also according to a view in perspective, the device of figure 4 to which it has been added also the support for a digital camera.
Figura 6.- Muestra, finalmente, otra perspectiva del dispositivo de la figura 4, al que se ha añadido además una pareja de fotodiodos para detección de fluorescencia.Figure 6.- Shows, finally, another perspective of the device of Figure 4, to which a further pair of photodiodes for fluorescence detection.
A la vista de las figuras reseñadas, especialmente de las figuras 1 a 3, puede observarse como el dispositivo que se preconiza está estructurado a partir de un armazón sólido (1) que configura a modo de una carcasa prismático-rectangular sustancialmente aplanada, en cuyo interior se aloja el diodo emisor de luz o LED (2) e inmediatamente encima de él el filtro de excitación (3), así como la fuente de alimentación para dicho diodo a través de un interruptor (4) preferentemente establecido en el borde frontal de la carcasa (1).In view of the figures outlined, especially from figures 1 to 3, it can be seen as the device that is recommended is structured from a solid frame (1) that configures as a housing substantially flattened rectangular-prismatic, in whose interior is the light emitting diode or LED (2) and immediately above it the excitation filter (3), as well as the power supply for said diode through a switch (4) preferably set on the front edge of the housing (1).
En situación de superposición con respecto al LED (2) y al filtro de excitación (3), en la carcasa se establece un rehundido (5) que permite colocar portas (6) con las muestras a visualizar.In a situation of overlap with respect to LED (2) and the excitation filter (3), the housing is set a recess (5) that allows to place slides (6) with the samples to visualize.
La estructura descrita se complementa con un brazo lateral (7) de configuración en "U", que emerge de la carcasa (1) a través de una de sus ramas laterales y que en su otra rama lateral, la superior, soporta al filtro de emisión (8).The structure described is complemented by a U-shaped side arm (7), which emerges from the housing (1) through one of its lateral branches and that in its other lateral branch, the upper one, supports the emission filter (8).
Estos filtros ópticos (3) y (8) existen en el mercado suministrados por distintos fabricantes, a la vez que los LED (2) constituyen igualmente elementos de mercado, comercializados por diversas firmas, permitiendo cubrir casi todo el espectro necesario para excitación de cromóforos.These optical filters (3) and (8) exist in the market supplied by different manufacturers, while the LED (2) also constitute market items, marketed by various firms, allowing to cover almost the entire spectrum necessary for chromophores excitation.
El diodo emisor de luz o LED (2) y los filtros (3) y (8), se encuentran situados a una distancia óptima para la visualización del material biológico.The light emitting diode or LED (2) and filters (3) and (8), are located at an optimal distance for the visualization of the biological material.
De acuerdo con el ejemplo de realización práctica representado en la figura 1, es posible la utilización de varios LED, en cuyo caso el brazo (7) constituye un elemento de soporte para respectivos filtros de emisión (8-8'), existiendo igualmente en el interior de la carcasa también respectivos filtros de excitación (3), en cuyo caso el brazo (7) es movible mediante guías apropiadas sobre la carcasa (1), en orden a poder efectuar combinaciones entre dichos LED, filtros de excitación y filtros de emisión, para la visualización de todo el espectro de colores fluorescentes mediante la utilización de filtros y diodos adecuados.According to the embodiment example practice represented in figure 1, it is possible to use several LEDs, in which case the arm (7) constitutes an element of support for respective emission filters (8-8 '), existing also inside the housing also respective excitation filters (3), in which case the arm (7) is movable by appropriate guides on the housing (1), in order to to be able to make combinations between said LEDs, filters excitation and emission filters, for viewing all the spectrum of fluorescent colors by using filters and suitable diodes.
A partir de esta estructuración básica, correspondiente al modo de realización más simple de la invención, es factible incrementar dimensionalmente la superficie en planta del armazón (1), tal como muestra la figura 4, para disponer sobre el mismo de un difusor de CO_{2} (9), alimentado por la correspondiente conducción de dicho gas (10), de los normalmente utilizados para anestesiar adultos de Drosophila, como ya se ha mencionado con anterioridad.From this basic structuring, corresponding to the simplest embodiment of the invention, it is feasible to dimensionally increase the planar surface of the frame (1), as shown in Figure 4, so as to have a CO_ diffuser thereon. 2} (9), fed by the corresponding conduction of said gas (10), of those normally used to anesthetize adults of Drosophila , as already mentioned above.
También se ha previsto la posibilidad de que en el dispositivo participe una cámara para fotografía de los resultados de la visualización, preferentemente una cámara digital, en cuyo caso el armazón (1) cuenta además con un segundo brazo (11), contrapuesto al brazo (7) anteriormente citado, de configuración similar pero de mayor altura, incorporando dicho brazo (11) en su extremidad libre un casquillo (12) o elemento similar, para fijación de la cámara, de manera que su objetivo quede alineado con el eje óptico pasante por los filtros de excitación (3) y de emisión (8), como se observa en la figura 5.The possibility that in the device participates a camera to photograph the display results, preferably a digital camera, in which case the frame (1) also has a second arm (11), as opposed to the arm (7) mentioned above, of similar configuration but of greater height, incorporating said arm (11) at its free end a bushing (12) or element similar, for fixing the camera, so that its objective is aligned with the through optical axis through the filters excitation (3) and emission (8), as seen in Figure 5.
Finalmente y tal como muestra la figura 6, se ha previsto también la posibilidad de incorporar al dispositivo al menos un foto-diodo (13), dos en el ejemplo de realización práctica de dicha figura 6, en correspondencia con los dos filtros emisores (8-8') para detección de fluorescencia en la misma pieza soporte (7) en la que se encuentran implantados los citados filtros de emisión (8-8').Finally, as shown in Figure 6, it has also provided for the possibility of incorporating the device into minus one photo diode (13), two in the example of practical realization of said figure 6, in correspondence with the two emitting filters (8-8 ') for detection of fluorescence in the same support piece (7) in which they are found the aforementioned emission filters implanted (8-8 ').
Claims (8)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES200603288A ES2330068B1 (en) | 2006-12-27 | 2006-12-27 | DEVICE FOR VISUALIZATION OF FLUORESCENT BIOLOGICAL MATERIAL. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES200603288A ES2330068B1 (en) | 2006-12-27 | 2006-12-27 | DEVICE FOR VISUALIZATION OF FLUORESCENT BIOLOGICAL MATERIAL. |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| ES2330068A1 true ES2330068A1 (en) | 2009-12-03 |
| ES2330068B1 ES2330068B1 (en) | 2010-08-30 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES200603288A Expired - Fee Related ES2330068B1 (en) | 2006-12-27 | 2006-12-27 | DEVICE FOR VISUALIZATION OF FLUORESCENT BIOLOGICAL MATERIAL. |
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| Country | Link |
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| ES (1) | ES2330068B1 (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4106438A (en) * | 1976-09-03 | 1978-08-15 | Nelson Roger K | Method of handling Drosophila |
| US20030230728A1 (en) * | 2002-06-13 | 2003-12-18 | Zhengshan Dai | Multiwavelength transilluminator for absorbance and fluorescence detection using light emitting diodes |
| US20040081621A1 (en) * | 2002-01-18 | 2004-04-29 | Frank Arndt | Optical imaging method, and an apparatus for optical imaging |
| WO2006062895A2 (en) * | 2004-12-06 | 2006-06-15 | Cambridge Research And Instrumentation, Inc. | Systems and methods for in-vivo optical imaging and measurement |
-
2006
- 2006-12-27 ES ES200603288A patent/ES2330068B1/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4106438A (en) * | 1976-09-03 | 1978-08-15 | Nelson Roger K | Method of handling Drosophila |
| US20040081621A1 (en) * | 2002-01-18 | 2004-04-29 | Frank Arndt | Optical imaging method, and an apparatus for optical imaging |
| US20030230728A1 (en) * | 2002-06-13 | 2003-12-18 | Zhengshan Dai | Multiwavelength transilluminator for absorbance and fluorescence detection using light emitting diodes |
| WO2006062895A2 (en) * | 2004-12-06 | 2006-06-15 | Cambridge Research And Instrumentation, Inc. | Systems and methods for in-vivo optical imaging and measurement |
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| Publication number | Publication date |
|---|---|
| ES2330068B1 (en) | 2010-08-30 |
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