GB2262163A - Fibre optic imaging bundle on which cells may be grown - Google Patents
Fibre optic imaging bundle on which cells may be grown Download PDFInfo
- Publication number
- GB2262163A GB2262163A GB9105010A GB9105010A GB2262163A GB 2262163 A GB2262163 A GB 2262163A GB 9105010 A GB9105010 A GB 9105010A GB 9105010 A GB9105010 A GB 9105010A GB 2262163 A GB2262163 A GB 2262163A
- Authority
- GB
- United Kingdom
- Prior art keywords
- optical assembly
- light
- light source
- cells
- growth
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000003384 imaging method Methods 0.000 title claims abstract description 8
- 239000000835 fiber Substances 0.000 title description 8
- 230000012010 growth Effects 0.000 claims abstract description 10
- 239000010453 quartz Substances 0.000 claims abstract description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 4
- 230000000694 effects Effects 0.000 claims abstract description 3
- 230000003287 optical effect Effects 0.000 claims description 13
- 230000005540 biological transmission Effects 0.000 claims description 5
- 230000010261 cell growth Effects 0.000 claims description 4
- 238000000034 method Methods 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 238000010521 absorption reaction Methods 0.000 claims description 2
- 230000001413 cellular effect Effects 0.000 claims description 2
- 230000001419 dependent effect Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- 238000002406 microsurgery Methods 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 claims 4
- 238000004113 cell culture Methods 0.000 claims 2
- WFKWXMTUELFFGS-UHFFFAOYSA-N tungsten Chemical compound [W] WFKWXMTUELFFGS-UHFFFAOYSA-N 0.000 claims 2
- 229910052721 tungsten Inorganic materials 0.000 claims 2
- 239000010937 tungsten Substances 0.000 claims 2
- 210000003527 eukaryotic cell Anatomy 0.000 claims 1
- 229910052736 halogen Inorganic materials 0.000 claims 1
- 150000002367 halogens Chemical class 0.000 claims 1
- 239000007788 liquid Substances 0.000 claims 1
- 210000001236 prokaryotic cell Anatomy 0.000 claims 1
- 230000000087 stabilizing effect Effects 0.000 claims 1
- 238000007789 sealing Methods 0.000 abstract description 2
- 239000007787 solid Substances 0.000 abstract 1
- 238000001228 spectrum Methods 0.000 description 5
- 238000004458 analytical method Methods 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- 238000010871 transoral laser microsurgery Methods 0.000 description 2
- 230000009102 absorption Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 239000004568 cement Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000001427 coherent effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000012538 light obscuration Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 1
- 230000002123 temporal effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B6/00—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings
- G02B6/04—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings formed by bundles of fibres
- G02B6/06—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings formed by bundles of fibres the relative position of the fibres being the same at both ends, e.g. for transporting images
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
-
- 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
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B18/00—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body
- A61B18/18—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves
- A61B18/20—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves using laser
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
- G01N2015/144—Imaging characterised by its optical setup
Landscapes
- Chemical & Material Sciences (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Plasma & Fusion (AREA)
- Sustainable Development (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Optics & Photonics (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
A whole cell sheet photometer has means for the growth of living cells e.g. in conditions of sterile spreading cell and tissue culture directly on the surface d of fibre-optic imaging bundle c the other end being bonded to a solid state area array photodetector a. Also shown are pelltier effect thermoelectric device b, inner chamber wall e for containing cells, secondary chamber f, lid g with quartz window, inlet and outlet ports h, self-sealing ports i, micro-stepping motor j and light source k. <IMAGE>
Description
This invention relates to an apparatus for examining cells either in the living state or histologically fixed, by visual inspection, and by other biochemical analytical methods which depend on the generation, modulation or absorption of light in the visible, uv and ir regions of the electromagnetic spectrum.
Spectrophotometers are well known instruments in the field of biochemistry which comprise a source of mono-chromatic light, a collimator a sample chamber and a recording output. Spectrophotometers are ideally suited to the analysis of macroscopic samples, usually in fluid form. Although successful attempts have been made to perform spectrophotometric analysis on single cells the methods employed so far and to date have proved both cumbersome and intricate to implement.
According to the present invention there is provided a means to perform the full range of spectrophotometrically dependent analyses on living cells in conditions of continuous spreading culture and to record the results continuously allowing furthermore the simultaneous (or entirely separate) recording of the visual data on changing cell morphology and dynamic ultrastructure changes by means of video micrography.
There is therefore provided a chamber and substrate for cell attachment and growth, a source of variably monochromatic light or white light, a means for collimating the light and guiding it through the cells under inspection, a means for first sensing and then recording the data on light transmission and other light associated phenomena arising from the presence of the cells and a means for moving the chamber in which the cells are growing with respect to the light source, which, where a laser is the light source, might simply be by means of a movable laser beam, so that individual sub sections of the whole attached cell growing area could be examined individually and/or be operated on by laser microsurgery.
A particular embodiment of the invention will now be described by way of example with reference to the accompanying drawings in which:
Fig 1: shows in side view the assembly a:is the mounting for an electronic area or linear array light detector such as a cCD or CID, most likely an area array bis a pelltier effect thermoelectric device for temperature stabilization of the chip c::is fibre optic bonded with optical cement to the full extent of CCD array, which may be a straight fibre optic imaging bundle or a tapered imaging bundle either of increasing or decreasing taper to the sample, in this instance decreasing taper, d)is polished "growth substrate" surface of the fibre-optic which is capable of being treated with a permanent or replenishible transparent coating which facilitates cell attachment and spreading - by the adsorption to this surface of polyanionic materials to this surface for example.
e):is the inner chamber wall for containing cells within the confines of the end surface of the fibre-optic when initially applied by pipette as a cell suspension f):larger secondary chamber with volume of growth medium which is capable of being temperature regulated, gas and pH stabilized and if necessary replenished continuously or batchwise.
g):lid to chamber with quartz window to allow transmission of full spectrum h): inlet and outlet ports for chamber perfusion when continuous circulation of medium is in operation i):self sealing ports for injection of test compounds j)micro-stepping motor attachable either to sample assembly stage or to collimated light source k):light source, laser sample beam and reference or non-coherent, diffuse or collimated mono-chromatic or polychromatic.
Referring to the drawing, the assembly comprises a temperature, gas and nutrient controlled environment for the growth of cells on a substrate which is the free surface of a fibre-optic imaging bundle which may be straight or tapered, but which is probably (for maximum resolution) of decreasing taper to the sample, the size of the fibres being selected so that the resolution of the image, of for instance cells growing or otherwise attached to the free surface, which image is transmitted to the electronic light detector at the bonded end will be maximised.The choice of fibre size is further governed by the reguirement to visualise with maximum resolution cellular phenomena which are associated with and can be correlated to observable variations in light extinction, absorption, transmission, refraction, luminescence and stimulated fluorescence which result from the particular spectrophotometric or luminometric protocols being used and so that the results can be monitored and recorded both at a high resolution within each cell and over an extended area of contiguous cell sheet.
In order to maximize resolution the size of the fibres in the fibre optic are chosen to match the size of the pixels in the detector array, with greater resolution - at the expense of overall size of possible cell growing area at the sample end - being made possible by tapering the bundle from the size determined by the array area size at the detector end to a desired smaller size at the growth end.
It can be deduced that resolution possible with this type of assembly is increased by reducing the size of the pixels in the detector array employed; it is further possible to increase resolution for a given size of pixel by tapering the optical fibres from their size at the chip-fibre interface which should match the pixel dimensions as closely as possible, to an arbitrarily smaller fibre size at the sample end the limits on this size being practically imposed by limitations on fibre optic drawing, and the wavelength of light.
In order to facilitate growth of cells, the sample growth end of the fibre-optic is an integral component of a growth chamber whose parameters of temperature, pH and nutrient composition can be both closely controlled and continuously monitored.
The chamber maintains a sterile environment for the long term continuous culture of cells attached to the fibre-optic growth end surface, and a quartz window is provided in the lid of the chamber to facilitate the transmission of all wavelengths of u.v., i.r. and visible radiation of interest.
In the instances that the apparatus is to be used as an ultra sensitive recording luminometer or fluorimeter, the temperature of the detector array is stabilized at a suitably low temperature to allow longer electronic exposure (integration) times by reducing interfering thermal background noise.
In order to allow small areas to be individually monitored for purposes of analysis and microsurgery, a micro-stepping motor may be incorporated in the assembly such that the growth stage is moved with respect to a stationary laser source or other highly collimated spectrophotometric source and reference beams. For laser microsurgery the laser beam itself may be manoeuverable.
Because the detector can be made very sensitive to signal over noise by thermoelectrically cooling the array, if a suitably intense white light source is used very rapid scans of the whole spectrum can be recorded in sequential conveniently short exposure frames; if the spectrum is divided into 10,000 bandwidths and each bandwidth is exposed for lOOusecs. the whole spectrum is scanned in 1 second which allows monitoring in a second by second time-lapse sequence of dynamic variations (within this temporal resolution) inside many different cells side by side simultaneously. Furthermore a time-lapse record of the data at any particular wavelength can be extracted, as can be a record of the dif; rence between any two wavelength bandwidth With modification the apparatus lends itself to a unique form of surface plasmon resonance recorder.
Claims (1)
- CLAIMS:1) A whole cell sheet visualiser (hereinafter termed "The Optical Assembly") comprising a fibre-optic imaging bundle on the free polished end surface of which living cells may be grown directly, e.g. in sterile cell culture conditions, the other end of which is optically bonded to a sensitive electronic imaging device, preferably an area array CCD or CID.2) An Optical Assembly as in claim 1) above wherein means are provided for light of any desired wavelength to be transmitted through the sample which is growing or has grown on the growth face of the fibre-optic.3) An Optical Assembly as in the above claims in which means is provided for the electronic light detecting array to be operated under computer control and the data stored for viewing or processing 4) An Optical Assembly as in the above claims in which means are provided for the detector array to be temperature stabilised and controlled e.g. by means of thermoelectric devices such as Pelltier Effect modules, or Stirling Cycle modules or where purely cryogenic operation is desired, liquid gas dewar cryostats.5) An Optical Assembly as in the above claims in which the light source may be a tungsten or tungsten/quartz halogen incandescent white light source which may be narrowed to a desired bandwidth by means of a prismatic assambly or by a variable diffraction grating or which may be a fixed wavelength or tunable laser source 6) An Optical Assembly as in claim 5) in which the light source may be suitably split to provide multiple sources of highly collimated source and reference beams for spectrophotometric and spectrofluorimetric determinations at a single point or multiple points on the cell sheet which may equally be the whole cell growth area analysed simultaneously.7) an Optical Assembly as in the above claims which is provided with a means for reorienting and monitoring the reorientation of highly collimated light beams for the purposes of microsurgery at the cellular level.8) An Optical Assembly as in the above claims in which a micr-stepping motor is provided where necessary to assist in orientation of sample with respect to light source.9) An Optical Assembly as in the above claims which is provided with a means for the sterile growth of eukaryotic or prokaryotic cells under conditions of continuous cell culture, with the means for monitoring and stabilizing the nutrient and gaseous exchange environment and pH and means for introducing test quantities of experimental materials into the cell growth chamber.10j An Optical Assembly -s in the above claims which can be used by ow some or all of the above features as a real time and recording imaging photometer with a wide field of view enabling an arbitrarily large expanse of living cell sheet to be examined simultaneously according to the methods of photometery, luminometery, scanning spectrophotometery, spectrofluorimetery and other techniques dependent on the differential absorption, transmission, diffraction, refraction fluorescence and generation of light in the particular experimental protocol of choice.l) An Optical Assembly substantially as described herein with reference to the accompanying drawings.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9105010A GB2262163B (en) | 1991-03-09 | 1991-03-09 | Whole cell sheet photometer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB9105010A GB2262163B (en) | 1991-03-09 | 1991-03-09 | Whole cell sheet photometer |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| GB9105010D0 GB9105010D0 (en) | 1991-04-24 |
| GB2262163A true GB2262163A (en) | 1993-06-09 |
| GB2262163B GB2262163B (en) | 1995-12-20 |
Family
ID=10691284
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| GB9105010A Expired - Fee Related GB2262163B (en) | 1991-03-09 | 1991-03-09 | Whole cell sheet photometer |
Country Status (1)
| Country | Link |
|---|---|
| GB (1) | GB2262163B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6846635B1 (en) | 1999-07-30 | 2005-01-25 | Large Scale Proteomics Corp. | Microarrays and their manufacture |
| WO2013030246A1 (en) * | 2011-08-30 | 2013-03-07 | General Electric Company | System for optical based delivery of exogenous molecules to cells |
| US8778682B2 (en) | 2011-08-30 | 2014-07-15 | General Electric Company | Optical based delivery of exogenous molecules to cells |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6653151B2 (en) | 1999-07-30 | 2003-11-25 | Large Scale Proteomics Corporation | Dry deposition of materials for microarrays using matrix displacement |
| CN107299055A (en) * | 2017-08-04 | 2017-10-27 | 西安捷骋仪器仪表有限公司 | A kind of intelligent program-controlled quantitative sealing machine |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1389247A (en) * | 1972-03-03 | 1975-04-03 | Dittrich W | Device for counting and characterising small particles |
| EP0209489A1 (en) * | 1985-07-01 | 1987-01-21 | Prutec Limited | Waveguide to be used as optical probe in multiple internal reflection spectroscopic analysis |
-
1991
- 1991-03-09 GB GB9105010A patent/GB2262163B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1389247A (en) * | 1972-03-03 | 1975-04-03 | Dittrich W | Device for counting and characterising small particles |
| EP0209489A1 (en) * | 1985-07-01 | 1987-01-21 | Prutec Limited | Waveguide to be used as optical probe in multiple internal reflection spectroscopic analysis |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6846635B1 (en) | 1999-07-30 | 2005-01-25 | Large Scale Proteomics Corp. | Microarrays and their manufacture |
| WO2013030246A1 (en) * | 2011-08-30 | 2013-03-07 | General Electric Company | System for optical based delivery of exogenous molecules to cells |
| US8778682B2 (en) | 2011-08-30 | 2014-07-15 | General Electric Company | Optical based delivery of exogenous molecules to cells |
Also Published As
| Publication number | Publication date |
|---|---|
| GB2262163B (en) | 1995-12-20 |
| GB9105010D0 (en) | 1991-04-24 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PCNP | Patent ceased through non-payment of renewal fee |
Effective date: 19960320 |
|
| PCNP | Patent ceased through non-payment of renewal fee |
Free format text: DELETE IN JOURNAL 5613, PAGE 5584 CEASED IN ERROR |
|
| PCNP | Patent ceased through non-payment of renewal fee |
Effective date: 19980309 |