Advanced Light Microscopy

Our advanced light microscopy facility is home to:

  • Two scanning-laser confocals with incubation and autofocus (Zeiss LSM80 with Airyscan SR; Leica Stellaris 5 with resonant scanner, super-Z Galvo, TauSense, Lightning SR)
  • A spinning-disk confocal microscope with incubation chamber (Crest X-lightv3 with optosplit and dual cameras)
  • A deconvolution fluorescence imaging platform (DeltaVision2)
  • A state-of-the-art fully automated stereozoom microscope (Zeiss AxioZoom V16)
  • Several widefield epifluorescence systems
  • A LUMICKS C-Trap optical tweezers–fluorescence microscope.

LUMICKS C-Trap optical tweezers–fluorescence microscopy system

An automated single-molecule optical-trap manipulation and super-resolved fluorescence imaging microscope

The Lancaster Biophysics Facility houses a state-of-the-art C-TrapⓇ Dual Optical Tweezers, Fluorescence and Confocal Microscopy system, enabling advanced single-molecule biophysics experiments at the interface of physics, biology, and chemistry. This powerful instrument was acquired with generous support from BBSRC ALERT 2024, significantly enhancing the facility’s capacity to perform world-leading research in molecular and cellular biophysics.

The C-Trap at Lancaster is available to researchers across Lancaster University and to external collaborators, providing a unique capability for the UK biophysics community. Training and experimental support are available to help users design and execute experiments that fully exploit the system’s advanced features.

Contact:

Nick Robinson (n.robinson2@lancaster.ac.uk)

Alexandre Benedetto (a.benedetto@lancaster.ac.uk)

Jayde Whittingham-Dowd (j.whittingham-dowd@lancaster.ac.uk)

The C-Trap integrates high-resolution dual optical tweezers, confocal-based fluorescence microscopy, label-free force measurements, and advanced multi-channel microfluidics into one seamless platform. This unique combination enables researchers to manipulate and directly observe single molecules in real time with nanometer spatial resolution and millisecond temporal resolution. Users can simultaneously measure mechanical forces and monitor conformational changes or binding events, providing unprecedented insight into dynamic molecular processes.

Applications span a broad range of research areas, including:

  • DNA–protein interactions (for example transcription, replication, repair, and chromatin remodeling)
  • RNA structure, folding, and RNA–protein interactions
  • Chromosome structure, dynamics, and associated protein complexes
  • Protein folding and unfolding
  • Mechanobiology by probing the role of mechanical forces in molecular and cellular processes
  • Biomolecular condensate formation and rheology
  • Biopolymer mechanics such as DNA, RNA, and cytoskeletal filaments
  • Drug-target interaction studies at the molecular scale with nuclear extracts

By combining precise force manipulation with correlative fluorescence imaging, the C-Trap enables experiments that would be impossible with conventional microscopy or biophysical techniques alone. Researchers can trap and position individual molecules, introduce reagents via microfluidics, and monitor their behaviour in a controlled environment, all within a single experiment.

  • Microscope type: LUMICKS C-Trap Dymo300
  • Confocal: diffraction limited
  • Fluorescence: 3 lasers: 488, 561 & 638nm
  • Detectors: single-photon APDs for confocal & back-illuminated CMOS camera for Brightfield
  • Objectives: Nikon CFI plan APO 60x/1.2W
  • Stage: XYZ-encoded automated
  • Optical trap: 20W trapping laser at 1064nm
  • Microfluidics: u-Flux system with 5 input lanes under laminar flow and 1 outlet.
  • Environmental control: closed-loop temperature control ambient -45C +/-0.2C
  • Software: Lakeview

Zeiss LSM 880 with Airyscan Confocal Microscope

Zeiss LSM 880 confocal microscope with Airyscan module for near super-resolution imaging. This fully automated microscope is used for optical sectioning, tiling and time lapse experiments for samples on microscope slides, petri dishes or multi-well plates. The system is fully enclosed in an environmental chamber allowing control of temperature and CO2 gas delivery during live-cell imaging. The Airyscan module opens up further applications such as imaging of individually labelled molecules or viral infection of cells while also allowing improved resolution for live-cell imaging without increased phototoxicity or photo-bleaching.

  • Microscope stand: inverted Zeiss Axiobserver Z1
  • Confocal: LSM880 with Airyscan
  • Fluorescence: 4 lasers and 6 laser lines: 405, 458, 488, 514, 561 & 633nm
  • Detectors: 3 detection channels including a GaAsP detector for higher sensitivity.
  • Objectives: Zeiss 10x/0.3D, 20x/0.8D, 40x/0.95D, 40x/1.4I, 63x/1.4I
  • Stage: XYZ automated
  • Accessories: Definite focus module for autofocus
  • Environmental control: Temperature ambient to 42C, CO2, humidity
  • Software: Zen Black

Leica Stellaris 5 Confocal Microscope

Leica Stellaris 5 confocal microscope with integrated white light laser (WLL), and Power HyD S silicon detectors. The fully tuneable WLL allows tuning of excitation between 485nm and 695nm with 1nm precision, while imaging up to 8 laser lines with freely configurable detection up to 850nm. The Power HyD detectors enable TauSense Technology, giving access to fluorescence lifetime-based information and intensity data simultaneously. The setup also includes Lightening SR mode, an information extraction technology that provides near super resolution images in real-time. A Super-Z Galvo stage affords fast, accurate Z-scanning and vibration free imaging. The system provides full environmental control for live imaging.

  • Microscope Stand: inverted Leica DMi8 CS premium
  • Confocal: Stellaris 5 with resonant scanner
  • Fluorescence: 485-685nm tuneable white light laser & 405nm Diode Laser
  • Detectors: 4 HyD S silicon detectors, TauSense & lightning SR modes
  • Objectives: 10x/0.4ApoD, 20x/0.75ApoD, 40x/0.95ApoD, 63x/1.4ApoI, 100x/1.4ApoI (all CS2 rated)
  • Stage: XYZ automated, Super-Z Galvo
  • Environmental control: Temperature ambient to 42C, CO2, humidity
  • Software: LASX suite with Advanced 3D, colocalization, dye separation, spectral scanning, FRET & FRAP modules.

Crest X-light v3 Spinning-disk microscope

Funded by: NWCR, LU, Wellcome Trust, and BBSRC

Custom-built Metamorph-operated Crest X-light v3 spinning-disk system for fast volumetric multichannel fluorescence imaging of live samples. This system can acquire two channels onto one camera at 100fps at full resolution and another channel on a secondary camera. The Z-nanopiezo combined with the Metamorph streaming mode enables acquisition of 3 channels simultaneously at several volumes per second. Main applications include live 3D cilium and calcium imaging.

Alexandre Benedetto (a.benedetto@lancaster.ac.uk)

  • Microscope stand: inverted Zeiss Axiovert M200
  • Confocal: Crest X-light v3 spinning-disk (15,000rpm) with integrated optosplit
  • Fluorescence: 89North LDI-7 405, 445, 470, 520, 555, 640, and 730nm & separate 365nm solid-state LED
  • Detectors: Hamamatsu Flash 4 V3 and Flash 4 Lite CMOS cameras
  • Objectives: Zeiss 10x/0.3NA dry, Zeiss 40x/1.2NA water
  • Stage: automated XY-encoded LUDL stage with 3i Z-nanopiezo
  • Accessories: Hamamatsu Optosplit II, CherryTemp (5-50C fast heating/cooling)
  • Environmental control: Temperature ambient to 42C, CO2.
  • Software: Metamorph

DeltaVision 2 Widefield Deconvolution Microscope

The DeltaVision deconvolution imaging system, by Applied Precision (now GE Healthcare) can generate high-resolution images from low fluorescent signals. It is built on an Olympus IX71 inverted microscope with a xenon lamp light source and 3 laser lines for fluorescent, DIC and 3D imaging. The system comprises a 6-position emission filter wheel and two camera options for maximizing sensitivity or resolution. The addition of the environmental chamber allows time-lapsed live cell imaging. Imaging on this system works best with relatively thin specimens and is well suited for weakly fluorescent signals, small samples (yeast, bacteria) and samples (or dyes) sensitive to photo-bleach/ toxicity.

  • Microscope stand: inverted Olympus IX71
  • Fluorescence: 3 laser lines 406, 488, and 532nm , 300 watt Xenon lamp for epifluorescence 
  • Filter Set: DAPI, FITC, TRITC, YFP, CFP
  • Detectors: Photometrics CoolSnap HQ for resolution & Cascade II EM-CDD for sensitivity
  • Objectives: 60x /1.42ApoI and 100x/1.4ApoI.
  • Software: DeltaVision2

Zeiss Axio Zoom.V16 Automated Macroscope

Fully-automated stereo zoom microscope (XYZ, focus, shutters, filters, and acquisition) with a choice of 15 fluorescent LEDs for 4-colour fluorescence and brightfield imaging. With very long working distance high-aperture macro lenses, a fast sensitive 12Mpixel camera, and tiling capabilities, this system can rapidly acquire high-resolution 3D fluorescence, phase, DIC and extended-focus images of large and small live samples, from protozoans to mammalian organs. The long working distance also enables imaging of dissections and easy screening of fluorescently tagged live samples in petri dishes and multiwell plates.

  • Microscope stand: upright Zeiss AxioZoom V16
  • Fluorescence: CoolLED PE-4000, 365, 385, 405, 435, 460, 470,490, 500, 525, 550, 580, 595, 635, 740, & 770nm
  • Filter sets: 90 HE LED (DAPI/ FITC/ TRITC/Cy5), 91 HE LED (CFP/YPF/mCherry), 92 HE LED: (DAPI/GFP/mCherry), customisable using extra 6-position emission filter wheel
  • Detector: Zeiss AxioCam 512mono
  • Objectives: Plan NeoFluar Z 1.0x/0.25 (FWD 56mm), Plan NeoFluar Z 2.3x/0.57 (FWD 10.6mm, 0.5NA in an object field of 1.5mm)
  • Stage: XY automated, Z automated (nosepiece)
  • Software: Zen Blue 2.6 with deconvolution, stitching, extended focus etc.

Widefield Epi-fluorescent Systems

The facility has a number of widefield epifluorescence systems for routine fluorescence imaging including:

  • Upright Zeiss Axioscope A1 with CoolLED PE-300 fluorescence illumination and AxioCam mono camera
  • Upright Leica DMRXA2 XYZ motorized microscope with CoolLED PE-300 fluorescence illumination and pco Panda 4.2bi CMOS camera
  • Upright Nikon E600 microscope with AxioCam colour camera
  • Inverted Zeiss Axioinvert 35 with AxioCam mono camera for imaging live cells