BAIR equipment

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

Savvas Nikolaou

Nikki R. Paul

• Amato C et al (2019) WASP Restricts Active Rac  to Maintain Cells' Front-Rear Polarization. Currrent Biology; 29(24):4169-4182.e4

• Juin A  et al (2019 ) N-WASP Control of LPAR1 Trafficking Establishes Response to Self-Generated LPA Gradients to Promote Pancreatic Cancer Cell Metastasis. Developmental Cell; 51:431-445.e7

• Riley JS et al (2018) Mitochondrial inner membrane permeabilisation enables mtDNA release during apoptosis. EMBO Journal; 3;37(17)

• Román-Fernández Á et al (2018) The phospholipid PI(3,4)P2 is an apical identity determinant. Nat Communications; 28;9(1):5041

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.

Joe Hodgson

Nikki R. Paul

• Parvy JP et al (2019) The antimicrobial peptide defensin cooperates with tumour necrosis factor to drive tum our cell death in Drosophila. Elife; 30;8. pii: e45061

• Beaumatin F et al (2019) mTORC1 Activation Requires DRAM-1 by Facilitating Lysosomal Amino Acid Efflux. Molecular Cell; 76: 163-176 e8

• Gay DM et al (2019) Loss of BCL9/9l suppresses Wnt driven tumourigenesis in models that recapitulate human cancer. Nature Communications; 10: 723

• Johansson J et al (2019) RAL GTPases Drive Intestinal Stem Cell Function and Regeneration through Internalization of WNT Signalosomes. Cell Stem Cell; 24: 592-607 e7

• Rudzka DA et al (2019) Migration through physical constraints is enabled by MAPK-induced cell softening via actin cytoskeleton re-organization. Journal of Cell Science; 132: jcs224071

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

• Papalazarou V et al (2020). The creatine–phosphagen system is mechanoresponsive in pancreatic adenocarcinoma and fuels invasion and metastasis. Nature Metabolism; 2: 62-80

• Kugeratski FG et al (2019) Hypoxic cancer-associated fibroblasts increase NCBP2-AS2/HIAR to promote endothelial sprouting through enhanced VEGF signaling. Science Signaling; 12(567)

• Fort et al (2018) Fam49/CYRI interacts with Rac1 and locally suppresses protrusions. Nature Cell Biology; 20(10): 1159-1171

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

• Fort et al (2018) Fam49/CYRI interacts with Rac1 and locally suppresses protrusions. Nature Cell Biology; 20(10): 1159-1171

• Dornier E et al (2017) Glutaminolysis drives membrane trafficking to promote invasiveness of breast cancer cells. Nature Communications; 21;8(1):2255

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 

• Humpton TJ et al (2018)  p53-mediated adaptation to serine starvation is retained by a common tumour-derived mutant. Cancer Metabolism; 6:18

 

 

 

 

 

 

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.

• Novo D et al, (2018) Mutant p53s generate pro-invasive niches by influencing exosome podocalyxin levels. Nature Communications; 29;9(1):5069

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.

• Neidier S et al (2019) Signature for Early Progression in KRAS-Driven Lung Adenocarcinoma. Cancers (Basel); 29;11(5)

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.

• Juin A  et al (2019 ) N-WASP Control of LPAR1 Trafficking Establishes Response to Self-Generated LPA Gradients to Promote Pancreatic Cancer Cell Metastasis. Developmental Cell; 51:431-445.e7

• Birch JL et al (2018) A Novel Small-Molecule Inhibitor of MRCK Prevents Radiation-Driven Invasion in Glioblastoma. Cancer Research; 15;78(22):6509-6522

• Hernandez-Fernaud JR et al (2017) Secreted CLIC3 drives cancer progression through its glutathione-dependent oxidoreductase activity. Nature Communications; 15;8:14206

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.

 

 

 

 

 

 

www.phasefocus.com

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/

• Rooney N et al (2020) RUNX1 is a driver of renal cell carcinoma correlating with clinical outcome. Cancer Research; pii: canres.3870.2019

• Spender LC et al (2019) Preclinical Evaluation of AZ12601011 and AZ12799734, Inhibitors of Transforming Growth Factor beta Superfamily Type 1 Receptors. Molecular Pharmacology; 95: 222-234

• Neidler S et al (2019) Identification of a Clinically Relevant Signature for Early Progression in KRAS-Driven Lung Adenocarcinoma. Cancers 2019; 11: 600

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/

• Papalazarou V et al (2020). The creatine–phosphagen system is mechanoresponsive in pancreatic adenocarcinoma and fuels invasion and metastasis. Nature Metabolism; 2: 62-80

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

Savvas Nikolaou

Nikki R. Paul

• Amato C et al (2019) WASP Restricts Active Rac  to Maintain Cells' Front-Rear Polarization. Current  Biology; 29(24):4169-4182.e4

• Juin A  et al (2019 ) N-WASP Control of LPAR1 Trafficking Establishes Response to Self-Generated LPA Gradients to Promote Pancreatic Cancer Cell Metastasis. Developmental Cell; 51:431-445.e7

• Riley JS et al (2018) Mitochondrial inner membrane permeabilisation enables mtDNA release during apoptosis. EMBO Journal; 3;37(17)

• Román-Fernández Á et al (2018) The phospholipid PI(3,4)P2 is an apical identity determinant. Nature Communications; 28;9(1):5041

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.

Frederic Fercoq

Amanda Mcfarlane

The Zeiss Elyra 7 with Lattice SIM is a laser-based widefield microscope that permits rapid acquisition, coupling super-resolution microscopy with high speed. This microscope is already proving useful in a number of projects, including live-cell imaging of microtubule dynamics, intracellular trafficking, mitochondrial localisation and 3D imaging of Drosophila. In addition to Structured Illumination, this microscope is capable of fast optical sectioning in Apotome mode, and Single Molecule Localisation Microscopy (SMLM) using STORM or 3D-PALM. The recent SIM2 upgrade allows even faster acquisition and higher resolution processing. To date this is the only Elyra 7 microscope in Scotland and is already fostering collaborations between the Beatson Institute and other Universities.

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.

Frederic Fercoq

Amanda Mcfarlane

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.

• Martin et al (2018) Accepting from the best donor; analysis of long-lifetime donor fluorescent protein pairings to optimise dynamic FLIM-based FRET experiments. PLoS One; 13(1):e0183585

• Miller BW et al (2015) Targeting the LOX/hypoxia axis reverses many of the features that make pancreatic cancer deadly: inhibition of LOX abrogates metastasis and enhances drug efficacy. EMBO Molecular Medicine ;7(8):1063-76

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.

• Reid SE et al (2017) Tumor matrix stiffness promotes metastatic cancer cell interaction with the endothelium. EMBO Journal; 15;36(16):2373-2389

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.

• Román-Fernández Á et al (2018) The phospholipid PI(3,4)P2 is an apical identity determinant. Nature Communications; 28;9(1):5041

The Operetta is a high-throughput confocal microscope primarily used for high-content screening and phenotype analysis.

• Rushworth LK et al (2020) Repurposing screen identifies mebendazole as a clinical candidate to synergise with docetaxel for prostate cancer treatment. British Journal of Cancer; 122: 517–527

• Neidler S et al (2019) Identification of a Clinically Relevant Signature for Early Progression in KRAS-Driven Lung Adenocarcinoma. Cancers; 11: 600

• Unbekandt M et al (2018) Discovery of Potent and Selective MRCK Inhibitors with Therapeutic Effect on Skin Cancer. Cancer Research; 15;78(8):2096-2114
  

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.

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.

• Martin et al (2018) Accepting from the best donor; analysis of long-lifetime donor fluorescent protein pairings to optimise dynamic FLIM-based FRET experiments. PLoS One; 13(1):e0183585

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.

• Martin et al (2018) Accepting from the best donor; analysis of long-lifetime donor fluorescent protein pairings to optimise dynamic FLIM-based FRET experiments. PLoS One; 13(1):e0183585

• Miller BW et al (2015) Targeting the LOX/hypoxia axis reverses many of the features that make pancreatic cancer deadly: inhibition of LOX abrogates metastasis and enhances drug efficacy. EMBO Molecular Medicine;7(8):1063-76

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.

• SE et al (2017) Tumor matrix stiffness promotes metastatic cancer cell interaction with the endothelium. EMBO Journal; 15;36(16):2373-2389

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.

• Amato C et al (2019) WASP Restricts Active Rac to Maintain Cells' Front-Rear Polarization. Current Biology; 16;29(24):4169-4182.e4

• Rudzka D et al (2019) Migration through physical constraints is enabled by MAPK-induced cell softening via actin cytoskeleton re-organization. Journal of Cell Science 132: jcs224071

• Raniero E et al (2015) Ligand-Occupied Integrin Internalization Links Nutrient Signaling to Invasive Migration. Cell Reports; 20: 10(3):398-413

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/

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.

• Novo D et al, (2018) Mutant p53s generate pro-invasive niches by influencing exosome podocalyxin levels. Nature Communications; 29;9(1):5069

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.

• Neidier S et al (2019) Signature for Early Progression in KRAS-Driven Lung Adenocarcinoma. Cancers (Basel); 29;11(5)

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.

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.

Frederic Fercoq

Amanda Mcfarlane

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.

• Reid SE et al (2017) Tumor matrix stiffness promotes metastatic cancer cell interaction with the endothelium. EMBO Journal; 15;36(16):2373-2389

Please contact Saadia Karim.

s.karim@beatson.gla.ac.uk

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.

Please contact Tom Gilbey for information of the flow cytometry equipment and analysis software.

t.gilbey@beatson.gla.ac.uk

Please contact Colin Nixon for information of the Histology and Immunohistochemistry services and imaging.

c.nixon@beatson.gla.ac.uk