BAIR equipment

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

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

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.

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

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.

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

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.

• 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

Incucyte S3 (1/2/3)

The Incucyte S3 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 Incucyte S3 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 S3 systems are often used to monitor Scratch Wound 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/

• 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

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

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

• 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

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.

Frederic Fercoq

Amanda Mcfarlane

Elyra 7

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.

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.

Frederic Fercoq

Amanda Mcfarlane

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.

• 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

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.

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

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.

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

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.

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.

• 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

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.

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

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.

• 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

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.

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

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.

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

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.

Frederic Fercoq

Amanda Mcfarlane

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.

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

IVIS Spectrum / Pearl / Ultrasound

Please contact Saadia Karim.

s.karim@beatson.gla.ac.uk

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.

t.gilbey@beatson.gla.ac.uk

Histology

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

c.nixon@beatson.gla.ac.uk