BAIR equipment
- Confocal Microscopes
- Widefield Microscopes
- Timelapse Microscopes
- Super-resolution Microscopes
- Multiphoton Microscopes
- High-Content Analysis
- FLIM/FRET
- Specialised Techniques
- In Vivo Imaging
- Other Equipment
- Analysis
Confocal Microscopes
Confocal microscopy is a very powerful tool and has many advantages over wide-field fluorescent microscopy. The use of a pinhole enables the user to eliminate the out of focus light and permits optical sectioning of cell or tissue samples. BAIR supports a number of standard and super-resolution confocal systems, which are also coupled to advanced technologies such as FLIM/FRET, TIRF or in vivo imaging.
Zeiss LSM 880 with Airyscan
The Zeiss LSM 880 with Airyscan is a super-resolution-capable confocal microscope. The 'Airyscan' detector delivers increased resolution (1.7x) with high sensitivity at around 140nm laterally and 400nm axially. The 'Airyscan Fast' mode increases scanning speeds by a factor of 4. This allows users to image at a higher speed without sacrificing sensitivity or resolution. It has a full cage environmental chamber for live cell imaging and is also suitable for photoactivation, FRAP and FCS.
Airyscan Principle: https://www.youtube.com/watch?v=QjKrLTHGnc8
Airyscan Fast: https://www.youtube.com/watch?v=ln6VUmLzLLc
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Zeiss LSM 710
The Zeiss LSM 710 is equipped with the Zeiss Quasar detector configured with 2, 3 or 34 channels. Built around a Zeiss Axioimager upright stand the system is primarily for fixed samples. It is suitable for a wide range of applications including imaging of quadrupled labelled specimens, colocalisation of proteins, and 3D imaging experiments.
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Nikon A1R
A fully automated, highly sensitive confocal microscope system, the Nikon A1R is built around a Nikon Ti Eclipse microscope with (Perfect Focus System) PFS. The system has a full cage environmental chamber and is therefore suitable for live cell timelapse imaging.
The Nikon A1R has two scanning systems: Firstly, there is a conventional sawtooth scanning system – galvano, which is used for high-resolution imaging where speed is not essential. Secondly, there is a fast resonant scanner, which is used for applications where the need for speed outweighs the need for high resolution. The two scanning systems can be operated in rapid succession or simultaneously. This system is therefore also suitable for photoactivation and FRAP experiments.
Amelie Juin
Olympus FV1000
The Olympus FV1000 is a confocal system designed for high resolution confocal imaging of both fixed and living cells. The FV1000 is built around a BX81 inverted microscope and is equipped with a SIM scanner that enables synchronisation of specimen excitation and observation (while one laser stimulates the second laser simultaneously provides high resolution imaging). This makes the FV1000 suitable for FRAP, FLIP and photoactivation experiments.
Anh Hoang Le
Widefield Microscopes
The Beatson Institute is equipped with a wide range of microscopes for multiple applications. These can be used for imaging tissues, cells or even nanoparticles such as exosomes. Microscopy techniques can also be coupled and complementary to other advanced technologies such as proteomics and metabolomics.
Olympus BX51
The BX51 microscope is a widefield system available for bright field and fluorescence observation methods. It is a non-motorised upright microscope system which is used primarily for fixed histological tissue sections and cells. It is also used for fixed fluorescently labelled samples.
http://olympus.magnet.fsu.edu/primer/java/lightpaths/bx51fluorescence/index.html
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Nanosight LM10
The Nanosight LM10 allows rapid and accurate analysis of the size distribution and concentration of all types of nanoparticles from 10nm to 2000nm in diameter, depending on the instrument configuration and sample type. The nanosight can be used to detect exosomes and microvesicles which are secreted from cells and tissues.
Leica Laser Microdissection
Laser microdissection uses a laser to isolate specific microscopic regions from tissue samples. The system is built around a Leica DM6000B upright microscope stand. These samples can be further analysed using additional techniques such as proteomics and metabolomics.
Zeiss Microinjection
The microinjection system consists of a Zeiss Axio Observer A1 microscope equipped with an Eppendorf Femtojet and Inject man N12. This system is mainly used to inject cells with nucleic acids or therapeutics.
Timelapse Microscopes
BAIR supports a range of long-term live-cell imaging microscopes with both manual and automated setups and analysis software.
Nikon Long-Term Timelapse (1/3/4)
BAIR has three Nikon TE 2000 Live-Cell Timelapse microscope systems with temperature control, CO2 and the Perfect Focus System (PFS). These systems are frequently used to image cell migration or cell division using phase/brightfield over time, and can also be used to detect fluorescent dyes or proteins.
Phasefocus
The Phasefocus VL21 Live Cell Imaging System permits label-free long-term timelapse imaging of live-cells. The Phasefocus uses a quantitative phase imaging (QPI) technology known as Ptychography to detect cells. Subsequent analysis can be used to characterise cell migration, morphology and proliferation over time.
Incucyte Zoom (1/3/4)
The Incucyte Zoom Live-Cell Imaging systems are automated microscopes that fit within tissue culture incubators. This permits long-term analysis of 2D and 3D cultures in multiple culture plate formats. The Incucye Zoom systems can also be used to detect fluorescent proteins or dyes, which are often used to measure cell proliferation and apoptosis. In combination with the WoundMaker tool, the Incucyte Zoom systems are often used to monitor Scratch Would Cell Migration over time in a 96-well plate format.
https://www.essenbioscience.com/en/products/incucyte/
Incucyte S3
The Incucyte S3 Live-Cell Imaging system is an automated microscope that fits within a tissue culture incubator. This permits long-term analysis of 2D and 3D cultures in multiple culture plate formats. The Incucyte S3 system can also be used to detect fluorescent proteins or dyes, which are often used to measure cell proliferation and apoptosis. In combination with the WoundMaker tool, the Incucyte S3 can be used to monitor Scratch Would Cell Migration over time in a 96-well plate format.
The Incucyte S3 has additional software modules such as the Spheroid analysis module, which permits measurement of 3D tumouroids and organoids over time.
https://www.essenbioscience.com/en/products/incucyte/incucyte-s3/
Super-resolution Microscopes
Zeiss LSM 880 with Airyscan
The Zeiss LSM 880 with Airyscan is a super-resolution-capable confocal microscope. The 'Airyscan' detector delivers increased resolution (1.7x) with high sensitivity at around 140nm laterally and 400nm axially. The 'Airyscan Fast' mode increases scanning speeds by a factor of 4. This allows users to image at a higher speed without sacrificing sensitivity or resolution. It has a full cage environmental chamber for live cell imaging and is also suitable for photoactivation, FRAP and FCS.
Airyscan Principle: https://www.youtube.com/watch?v=QjKrLTHGnc8
Airyscan Fast: https://www.youtube.com/watch?v=ln6VUmLzLLc
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Zeiss Multiphoton 880
The Zeiss Multiphoton 880 with Airyscan is a super-resolution-capable confocal microscope. This system is in an upright configuration, and is suited to in vivo imaging of e.g. skull/brain, lung sections and abdominal organs with the use of vacuum windows.
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Elyra 7
Multiphoton Microscopes
Zeiss Multiphoton 880
The Zeiss Multiphoton 880 with Airyscan is a super-resolution-capable confocal microscope. This system is in an upright configuration, and is suited to in vivo imaging of e.g. skull/brain, lung sections and abdominal organs with the use of vacuum windows.
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Trimscope 1
The LaVision TRIM microscope is a high-resolution multiphoton system able to image small groups of cells down to the level of subcellular structure and generate three-dimensional reconstructions. It is capable of fast and sectioned FLIM, as well as detecting second-harmonic generation (SHG). The system has a heated stage and CO2 so is suitable for live-cell imaging.
This system has been used to image neurons of the central nervous system and epithelial cells within the crypts of the colon.
Trimscope 2
The LaVision TRIM microscope is a high-resolution multiphoton system able to image small groups of cells down to the level of subcellular structure and generate three-dimensional reconstructions. It is capable of fast and sectioned FLIM, as well as detecting second-harmonic generation (SHG).
This system is capable of in vivo imaging using e.g. abdominal or mammary windows. In addition, this system has dual channel FLIM detectors so can do simultaneous FLIM measurements of CFP and GFP-based biosensors.
High-Content Analysis
Opera Phenix
The Opera Phenix is a high-throughput confocal microscope primarily used for high-content screening and phenotype analysis. The system supports automated plate loading and image acquisition. The Opera Phenix is particulary advanced in 3D imaging of cells, organoids, spheroids and tissues. Coupled to Harmony/Columbus analysis software, the Opera Phenix can generate large amounts of data suitable for bioinformatics and machine learning.
Operetta
The Operetta is a high-throughput confocal microscope primarily used for high-content screening and phenotype analysis.
FLIM/FRET
Andor SD FLIM
The Andor Revolution XD is built around a Nikon Ti- inverted microscope. This system has a Yokogawa CSU-X spinning disc confocal unit with a disk speed that supports up to 2,000 frames per second. The Lambert Instruments FLIM Attachment for lifetime imaging microscopy is a dedicated system that allows rapid image acquisition and generation of lifetime images in the frequency domain. To achieve this the system uses a modulated light source and a modulated image intensifier as a detector.
FLIM WF/TIRF
This system has a LIFA: Lambert Instruments Fluorescence Attachment for lifetime imaging. It is attached to a Nikon TE-2000 microscope with PFS. The LIFA system calculates lifetimes based on the frequency domain method by using a modulated light source and a modulated image intensifier as detector. This Lambert system is coupled through our custom TIRF condenser to make it a FLIM-TIRF system for GFP. The system can also be used for widefield FLIM.
Trimscope 1
The LaVision TRIM microscope is a high-resolution multiphoton system able to image small groups of cells down to the level of subcellular structure and generate three-dimensional reconstructions. It is capable of fast and sectioned FLIM, as well as detecting second-harmonic generation (SHG). The system has a heated stage and CO2 so is suitable for live-cell imaging.
This system has been used to image neurons of the central nervous system and epithelial cells within the crypts of the colon.
Trimscope 2
The LaVision TRIM microscope is a high-resolution multiphoton system able to image small groups of cells down to the level of subcellular structure and generate three-dimensional reconstructions. It is capable of fast and sectioned FLIM, as well as detecting second-harmonic generation (SHG).
This system is capable of in vivo imaging using e.g. abdominal or mammary windows. In addition, this system has dual channel FLIM detectors so can do simultaneous FLIM measurements of CFP and GFP-based biosensors.
Specialised Techniques
Nikon TIRF
The facility has a total internal reflection fluorescence (TIRF) microscope built around a Nikon Eclipse inverted stand. TIRF is a powerful technique that uses an evanescent wave to excite fluorophores that are present at the bottom of the specimen adjacent to the glass water interface. The optical section that is excited is in the range of 100nm-150nm. It is often employed to study cellular membrane activities, the dynamics of actin, cellular adhesion, cell movement, single molecular events, vesicle and protein tracking. One of the advantages of TIRF is that the background and out of focus signals are dramatically reduced, which allows you to view very dim fluorescent events. This system is appropriate for live-cell fluorescent protein imaging.
PRIMO
The PRIMO device allows users to micropattern custom microenvironments for fixed- and live-cell experiments. The PRIMO is coupled to a fluorescent microscope and uses photopatterning technology to generate both 2D and 3D microfabricated functionalised structures.
https://www.alveolelab.com/our-products/primo-micropatterning/
Nanosight LM10
A Nanosight LM10 allows rapid and accurate analysis of the size distribution and concentration of all types of nanoparticles from 10nm to 2000nm in diameter, depending on the instrument configuration and sample type. The nanosight can be used to detect exosomes and microvesicles which are secreted from cells and tissues.
Leica Laser Microdissection
Laser microdissection uses a laser to isolate specific microscopic regions from tissue samples. The system is built around a Leica DM6000B upright microscope stand. These samples can be further analysed using additional techniques such as proteomics and metabolomics.
Zeiss Microinjection
The microinjection system consists of a Zeiss Axio Observer A1 microscope equipped with an Eppendorf Femtojet and Inject man N12. This system is mainly used to inject cells with nucleic acids or therapeutics.
In Vivo Imaging
Zeiss Multiphoton 880
The Zeiss Multiphoton 880 with Airyscan is a super-resolution-capable confocal microscope. This system is in an upright configuration, and is suited to in vivo imaging of e.g. skull/brain, lung sections and abdominal organs with the use of vacuum windows.
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Trimscope 2
The LaVision TRIM microscope is a high-resolution multiphoton system able to image small groups of cells down to the level of subcellular structure and generate three-dimensional reconstructions. It is capable of fast and sectioned FLIM, as well as detecting second-harmonic generation (SHG).
This system is capable of in vivo imaging using e.g. abdominal or mammary windows. In addition, this system has dual channel FLIM detectors so can do simultaneous FLIM measurements of CFP and GFP-based biosensors.
IVIS Spectrum / Pearl / Ultrasound
Please contact Saadia Karim.
Other Equipment
Tissue Culture Microscopes
BAIR supports a number of tissue culture microscopes throughout the Beatson Institute. All these microscopes can be used to visualise fluorescent proteins in cells, and are coupled to a PC and camera.
Flow Cytometry
Please contact Tom Gilbey for information of the flow cytometry equipment and analysis software.
Histology
Please contact Colin Nixon for information of the Histology and Immunohistochemistry services and imaging.
Analysis
The BAIR Centre for Analysis and Visualisation of Experimental data is a dedicated analysis suite which gives users access to high-performance computers and both licenced and open-source software.
Software includes:
- Fiji
- Imaris
- Icy
- Harmony
- Zen
- NIS Elements
- Incucyte
- Cell Profiler
- Metamorph
- Phasefocus
- Flowjo