Confocal and Super-resolution Microscopy Facility

Within the Bioimaging Centre in the School of Engineering and Materials Science we have the following two confocal microscope systems: a Nikon CSU-W1 SoRa spinning disk, and a Zeiss LSM710 Elyra PS.1 super resolution system purchased through the Institute of Bioengineering, as well as two standard epifluorescence microscopes and an incubator based Lumascope 720 for live cell imaging, and Chiaro nanoindenter mounted with Leica DMi8 for measuring mechanical parameters of cells and soft materials. We also have Nanosight LM20 for size distribution of particles, iBright FL1500 for western blots and nucleic acid gels, and Imaris 9.7.2 for imaging data analysis. The bioimaging facilities have contributed significantly to the publication output in SEMS and other institutes at QMUL. To apply for a training of any of our bioimaging facilities, please request through iLab.

 

Zeiss Super resolution LSM710 ELYRA PS.1

LSM 710 ELYRA PS.1 is a combination of an inverted laser scanning confocal microscopy 710 (LSM 710) with super resolution imaging microscopy ELYRA PS.1: photo activated localization microscopy (PALM)/Stochastic Optical Resolution Microscopy (STORM) and structured illumination microscopy (SIM). It is suitable for imaging targeting at 10-100nm resolution at single molecule level. In addition, Zen 2012 software, motorised X/Y stage, definite focus, TIRF mode make it capable of acquisition of tiling, multiple positions, time lapse, FRAP and FRET. It has a on stage environment chamber with a 35mm petri dish adaptor linked with a CO2/N2 controller for live cell imaging including hypoxic experiments. Furthermore, it comes with an offline data processing workstation.

Objectives:

  • EC Plan-Neofluar10x/0.3 M27,
  • EC Plan-Neofluar20x/0.5 M27,
  • Plan-Apochromat 63x/1.4 Oil DIC M27,
  • Plan-Apochromat 100x/1.46 Oil DIC M27

Lasers:

  • LSM 710: Diode laser 405nm (30mW), Ar/ML 458/488/514nm (35mW), HeNe 543nm (1mW), HeNe 633nm (5mW)
  • SIM/PALM: HR Diode 405nm (50mW), HR Diode 488nm (100mW), HR DPSS 561nm (100mW), HR Diode 642nm (150mW)

Cameras:

  • pco.edge sCMOS for SIM
  • Andor iXon DU 897 back-illuminated EMCCD for PALM/STORM

SOP Zeiss LSM 710 Elyra PS1 superresolution

Charge:

Zeiss Elyra Charge

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Nikon CSU-W1 SoRa Spinning Disk Confocal

Nikon SoRa

Nikon CSU-W1 SoRA Super-Resolution Spinning Disk Confocal scanning unit, couples the Nikon Ti2-E inverted microscope platform to the CSU-W1 SoRa system from Yokogawa. The system is equipped with two camera adapters, disk changer with two spinning disks (a 50 μm pinhole W1 spinning disk, and a 50 μm micro-lensed pinhole SoRa spinning disk), a full enclosure incubator with temperature, CO2 and humidity control for live imaging, FRAP and FRET. The system uses NIS-Elements software for data acquisition and image processing, and it comes with an offline data processing workstation.

Objectives:

  • CFI Plan Fluor 10x/0.3/WD 16mm,
  • CFI Plan Apochromat VC 20x/0.75/WD 1mm,
  • CFI Super Fluor 40xC/0.9/WD 0.34-0.26mm,
  • CFI Aprochromat TIFR 60xC/1.49/WD 0.13mm Oil

Lasers:

  • LuxX Diode lasers: 405nm (120mW), 445nm (100mW), 488nm (200mW), 515nm (100mW), 638nm (200mW)
  • Coherent OBIS LS laser 561nm (150mW)

Camera:

  • Two Photometrics Prime BSI back-illuminated sCMOS cameras

SOP Nikon CSU W1 SoRa

Charge:

Nikon SoRa Charge

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Leica DMI4000B Epifluorescence Microscope

Leica DMI4000B

Leica DMI4000B is an inverted epifluorescence widefield (WF) microscope suitable for bright field (BF), phase contrast (PH) acquisition and epifluroscence imaging with DAPI, FITC, TRITC, and Texas Red filters on fixed cell and tissue samples.

Objectives:

  • N PLAN 2.5x/0.07
  • HCX PL FLUOTAR 10x/0.3 PH1,
  • HCX PL FLUOTAR 20x/0.5 PH2,
  • HCX PL APO 40x/1.25-0.754 Oil,
  • HCX PL FLUOTAR 63x/1.25 Oil PH3

Camera:

  • LEICA DFC9000 GT sCMOS camera for epifluorescence and BF/PH imaging

SOP Leica DMI4000B

Charge:

Leica DMI4000B Charge

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Leica DMi8 Epifluorescence Microscope

Leica DMi8 Chiaro

Leica DMi8 is a semiautomatic inverted epifluorescence widefield (WF) microscope suitable for bright field (BF), phase contrast (PH) acquisition and epifluroscence 2D and 3D imaging with DAPI, GFP, Cy3, and Cy5 filters on fixed cell and tissue samples.

Objectives:

  • HC PL FLUOTAR 10x/0.32 PH1,
  • HC PL FL L 20x/0.40 CORR, PH1,
  • HC PL FL L 40x/0.60 CORR PH2,
  • HC PL FLUOTAR 63X/1.30 Oil PH3

Camera:

  • LEICA DFC9000 GT sCMOS camera for epifluorescence and BF/PH imaging

SOP Leica DMi8

Charge:

Leica DMi8 Chiaro charge

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Lumascope 720

Lumascope 720

Lumascope 720 is an automatic incubator based, inverted 3-colour (blue, green, red) epifluorescence microscope, with motorized XY stage and auto Z-focus. It is suitable for multiple positions, time lapse live imaging with both phase contrast and fluorescence 2D and 3D mode, up to 10 fps, or 30 fps with reduced frame size.

Objectives:

  • Motic EF-N Plan 4x/0.10 (not for imaging),
  • Olympus CAch N 10x/0.25,
  • Olympus LCAch N 20x/0.40,
  • Olympus LCAch N 40x/0.55

Camera:

  • CMOS camera for epifluorescence and PH imaging

SOP Lumascope 720

Charge:

Lumascope Charge

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Chiaro Nanoindenter

Chiaro Nanoindenter

Chiaro nanoindenter is the ideal nanoindentation instrument to explore the micro-mechanical properties of cells, spheroids or other small samples. Therefore, this instrument is purposely built to measure forces on cells and micro-materials while placed on an inverted microscope Leica DMi8. The Chiaro nanoindenter is capable of measuring many mechanical parameters. These include the Young’s Modulus (E), storage (E’) and loss moduli (E”). Moreover, both, static and dynamic indentation can be performed using this instrument, including creep, stress-relaxation and DMA.

SOP Chiaro Nanoindenter

Charge:

Chiaro Charge

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Nanosight LM20

Nanosight LM20

Nanosight LM20 with a LM10 laser unit (Class 1 diode 635 nm) characterizes the size distribution and concentration of all types of nanoparticles from 20nm-1000nm in solution by dynamically analysing the paths the particles take under Brownian motion over a suitable period of time.

SOP Nanosight LM20

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 iBright FL1500

                                           iBright FL1500

iBright FL1500 Imaging System supports the main imaging applications of fluorescent, chemiluminescent, and colorimetric western blots, in addition to fluorescent stained nucleic acid gels, fluorescent stained protein gels, colorimetric stained protein gels, and colorimetric membrane stains. 

Features of the iBright FL1500 Imaging System:
• 9.1 MP cooled CCD camera: high sensitivity and dynamic range to help enable the detection of subtle differences in samples
• Five fluorescence channels: multiplex and capture up to four proteins in a single blot for more meaningful and representative experiments
• Smart Exposure technology: provides rapid determination of optimal exposure time to help minimize the need to repeat exposures to acquire the desired signal
• Simple interface: clear layout of functions and features combined with a 12.1-inch capacitive touchscreen for a smooth imaging experience
• Advanced automated features: automatic sample rotation, automatic zoom, and automatic focus help streamline image capture
• 22.5 x 18.0-cm field of view: large field of view for high-throughput imaging (image up to four mini or two midi blots at a time)
• Green LED-based transilluminator: effectively excite popular DNA dyes such as ethidium bromide and Invitrogen SYBR Green dyes with an alternative to UV-based transilluminators
• Flexible connectivity: export captured images via ethernet connection, Wi-Fi (with optional accessory), USB, or directly to Connect cloud-based platform
• Multiple image analysis options: perform densitometry, quantitation, and normalization directly using the on-instrument software, or for more in-depth analysis use iBright Analysis Software (available in both desktop and cloud-based versions)

                      iBright FL1500 filters

SOP iBright FL1500

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Imaris 9.7.2 ----- Imaris Single Full with ClearView

Imaris 9.7.2

Imaris Single Full gives you complete power and flexibility of all Imaris functionalities at your fingertips. Visualization of complex 3/4D microscopy datasets with automated Spots and Surfaces detection and visualisation (100s of GBs), smart detection of complex objects, tracing of neurons, blood vessels (no lumen) or other filamentous structures, tracking including cell division detection, batch analysis and a wide range of customized analysis powered by MatLab or Python. Imaris integrates ClearView™ algorithms for Spinning Disk Confocal, Widefield Fluorescence, Brightfield, Laser Scanning Confocal and TIRF, allowing GPU-accelerated Deconvolution. 

Imaris Quickstart Tutorials

Imaris Reference Manual

For more information:

https://imaris.oxinst.com/products/imaris-single-full

Online learning centre: 

https://imaris.oxinst.com/learning/

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Publications

  • Chronopoulos A, Thorpe SD, Cortes E, Lachowski D, Rice AJ, Mykuliak VV, et al. (2020). Syndecan-4 tunes cell mechanics by activating the kindlin-integrin-RhoA pathway. Nat Mater 19(6), 669-678. doi: 10.1038/s41563-019-0567-1 Impactor factor: 38.663
  • Torres-Pérez JV, Naeem H, Thompson CL, Knight MM and Novak P (2020). Nanoscale mapping reveals functional differences in ion channels populating the membrane of primary cilia. Cellular Physiology and Biochemistry 54(1), 15-26. doi: 10.33594/000000202 Impactor factor: 5.11
  • Di Cio S, Iskratsch T, Connelly JT and Gautrot JE (2019). Contractile myosin rings and cofilin-mediated actin disassembly orchestrate ECM nanotopography sensing. Biomaterials 232. doi: 10.1016/j.biomaterials.2019.119683 Impactor factor: 12.121
  • Walker RV, Keynton JL, Grimes DT, Sreekumar V, Williams DJ, Esapa C, Wu D, Knight MM, Norris DP (2019). Ciliary exclusion of Polycystin-2 promotes kidney cystogenesis in an autosomal dominant polycystic kidney disease model. Nat Commun 10(1):4072. doi: 10.1038/s41467-019-12067-y Impactor factor: 12.121
  • Okesola BO, Wu Y, Derkus B, Gani S, Wu D, Knani D, Smith DK, Adams DJ, Mata A (2019) Supramolecular self-sssembly to control structural and biological properties of multicomponent hydrogels. Chem Mater 31(19):7883-7897. doi: 10.1021/acs.chemmater.9b01882 Impactor factor: 9.567
  • Li W and Wang W (2019). Membrane tension regulates syndecan-1 expression through actin remodelling. Biochim Biophys Acta Gen Subj 1863(11), 129413-129413. doi: 10.1016/j.bbagen.2019.129413 Impactor factor: 4.008
  • Fu S, Thompson CL, Ali A, Wang W, Chapple JP, Mitchison HM, Beales PL, Wann AKT and Knight MM (2019). Mechanical loading inhibits cartilage inflammatory signalling via an HDAC6 and IFT-dependent mechanism regulating primary cilia elongation. Osteoarthritis and Cartilage 27(7), 1064-1074. doi: 10.1016/j.joca.2019.03.003 Impactor factor: 4.68
  • Rowson DT, Shelton JC, Screen HRC and Knight MM (2018). Mechanical loading induces primary cilia disassembly in tendon cells via TGFβ and HDAC6. Scientific Reports 8(1), 11107. doi: 10.1038/s41598-018-29502-7 Impactor factor: 4.576
  • Gushchina S, Pryce G, Yip P, Wu D, Pallier P, Giovannoni G, Baker D, Bo X (2018). Increased expression of colony-stimulating factor-1 in mouse spinal cord with experimental autoimmune encephalomyelitis correlates with microglial activation and neuronal loss. Glia 66(10), 2108-2125. doi: 10.1002/glia.23464 Impactor factor: 5.984
  • Freeley M, Attanzio A, Cecconello A, Amoroso G, Clement P, Fernandez G, Gesuele F, Palma M (2018). Tuning the Coupling in Single-Molecule Heterostructures: DNA-Programmed and Reconfigurable Carbon Nanotube-Based Nanohybrids. Adv Sci 5(10), 1800596-1800596. doi: 10.1002/advs.201800596 Impactor factor: 15.84
  • Thorpe SD, Gambassi S, Thompson CL, Chandrakumar C, Santucci A, and Knight MM (2017). Reduced primary cilia length and altered Arl13b expression are associated with deregulated chondrocyte Hedgehog signaling in alkaptonuria. J Cell Physiol 232(9), 2407-2417. doi: 10.1002/jcp.25839 Impactor factor: 5.096
  • Gambassi S, Geminiani M, Thorpe SD, Bernardini G, Millucci L, Braconi D, et al. (2017). Smoothened-antagonists reverse homogentisic acid-induced alterations of Hedgehog signaling and primary cilium length in alkaptonuria. J Cell Physiol 232(11), 3103-3111. doi: 10.1002/jcp.25761 Impactor factor: 5.096
  • Thompson CL, Plant JC, Wann AK, Bishop CL, Novak P, Mitchison HM, Beales PL, Chapple JP and Knight MM (2017). Chondrocyte expansion is associated with loss of primary cilia and disrupted hedgehog signalling. Eur Cell Mater 34, 128-141. doi: 10.22203/eCM.v034a09 Impactor factor: 3.741
  • Schwarz N, Lane A, Jovanovic K, Parfitt DA, Aguila M, Thompson CL, da Cruz L, Coffey PJ, Chapple JP, Hardcastle AJ, Cheetham ME (2017). Arl3 and RP2 regulate the trafficking of ciliary tip kinesins. Human Molecular Genetics 26, 13(1), 2480–2492. doi: 10.1093/hmg/ddx143 Impactor factor: 5.1
  • Wu D, Lee S, Luo J, Xia H, Gushchina S, Richardson PM, Yeh J, Krügel U, et al. (2017). Intraneural injection of ATP stimulates regeneration of primary sensory axons in the spinal cord. Journal of Neuroscience 38(6) 1351-1365. doi: 10.1523/JNEUROSCI.1660-17.2017 Impactor factor: 5.673
  • Freeley M, Worthy HL, Ahmed R, Bowen B, Watkins D, Macdonald JE, Zheng M, Jones DD, et al. (2017). Site-Specific One-to-One Click Coupling of Single Proteins to Individual Carbon Nanotubes: A Single-Molecule Approach. J Am Chem Soc 139(49), 17834-17840. doi. 10.1021/jacs.7b07362 Impactor factor: 14.612
  • Thompson CL., Wiles, A., Poole CA. and Knight MM (2016). Lithium chloride modulates chondrocyte primary cilia and inhibits Hedgehog signaling. The FASEB Journal 30(2), 716-726. doi: 10.1096/fj.15-274944 Impactor factor: 4.966
  • Sliogeryte K, Thorpe SD, Wang Z, Thompson CL, Gavara N, and Knight MM(2016). Differential effects of LifeAct-GFP and actin-GFP on cell mechanics assessed using micropipette aspiration. J Biomech 49(2), 310-317. doi: 10.1016/j.jbiomech.2015.12.034 Impactor factor: 2.32

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To request a training via iLab

Cell and TissueLab Bioimaging Training

 

Contact

Prof Martin Knight
email: m.m.knight@qmul.ac.uk
Tel: +44 (0)20 7882 8868

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