Sartorius IncuCyte S3 Live-Cell Analysis System
The IncuCyte S3 system is ideal for long-term, automated live-cell analysis of up to 6 plates. The system resides inside an incubator which enables consistent maintenance of environmental controls at all times of the day and enables long-term experiments.
Images can be captured in HD phase and fluorescence (green and red) at 4x, 10x and 20x.
The IncuCyte software allows remote setup, data visualisation and data analysis. The basic version of the software provides capabilities for experiments such as cell counting, cell viability, cytotoxicity, cell cycle, immune cell, chemotaxis, whole well and dilution cloning. Additional experiments are available to purchase in agreement with the Facility.
Remote data analysis includes; (A) automatically acquire images over time, (B) use Incucyte® VesselView to view images of all locations in the vessel at once and quickly assess experimental results, plus zoom in on images of interest (C) automatically identify regions of interest via masks (D) generate presentation-ready timelapse graphs, and (E) view all 96- or 384-well kinetic trends at once with Incucyte® PlateGraph and export data to calculate EC50 or IC50 response values.
Features
Speed and Sensitivity In Mind
- CMOS camera and efficient optics enable long-term live-cell imaging and analysis
- 20 total core processors allowing for fast image processing and data analysis
Data Protection & Storage
- Includes 27.3 TB of RAID hard drive data storage, which can be further extended with the 32.7 TB Incustore
LCD Control Panel
- Monitor and perform basic status checks and operations from the front panel of the Incucyte S3 System controller unit
Software and Network Drives
- IncuCyte 2022B Rev2 – On workstations in WXG.90 or available via remote download.
Location
Imaging Suite: Room WXG.89
For more information, please contact microscopyfacility@contacts.bham.ac.uk
Zeiss LSM780 Confocal Microscope
The LSM 780 scanner has high sensitivity and is equipped with a 32 element galanium arsenide phosphide (GaAsP) Quasar detector, plus 2 standard PMTs and 1 transmitted light T-PMT. Equipped with a tuneable AOTF and capable of imaging up to 6 channels in multitrack configuration. The AxioObserver.Z1 inverted stand, is equipped with an environmental chamber with CO2 for live cell imaging. Focus stabilisation is through software and hardware based mechanisms. Interchangeable stage inserts for slides, chamber slides, 35mm dishes and multiwall plates. Image capture in 3D and 4D, plus Lambda scanning, and spectral un-mixing capability.
Optics
| Objective lens | Magnification / NA / Immersion | Contrast Method |
|---|
|
C-ACHROMAT
|
10x / 0.45 / Water
|
|
|
LD LCI Plan-NEOFLUAR
|
25x / 0.8 / Water or Oil
|
DIC II
|
|
Plan-ACHROMAT
|
40x / 1.2 / Water
|
DIC III
|
|
Plan-APOCHROMAT
|
63x / 1.2 / Water
|
DIC III
|
|
Plan-APOCHROMAT
|
63x / 1.4 / Oil
|
DICIII
|
|
Plan-APOCHROMAT
|
100x / 1.4 / Oil
|
DIC III
|
Epifluorescence filter sets
| Filter set | Ex BP nm | Em BP nm |
|---|
|
Fs49 wf
|
365
|
445/50
|
|
Fs38 wf
|
470/40
|
525/50
|
|
Fs43 wf
|
545/25
|
635/70
|
Laser Lines
- Lasos 30mW Diode 405nm
- Lasos 25mW LGN3001 multiline Argon, 458, 488, 514nm
- Lasos 20mW (DPSS 561) 561nm
- Lasos 2mW HeNe 594nm
- Lasos 5mW HeNe 633nm
Experiment Modes
- Z stack
- Multi-positions
- Time lapse
- Tile scanning / image stitching of large samples
- Bleaching
Software and Network Drives
- Zen 2.1 allows for multidimensional experiment design in 2D, 3D, and 4D
- PC, HPZ840 4TB 3 RAID Hard drives and 64GB of memory.
- Desk top script connections to personal offline storage folders
Location
Imaging Suite, Room WXG86
Zeiss LSM880 Confocal Microscope with Airyscan Fast
The LSM 880 scanner has high sensitivity and is equipped with a 32 element galanium arsenide phosphide (GaAsP) Quasar detector, plus 2 standard PMTs and 1 transmitted light T-PMT. Equipped with a tuneable AOTF and capable of imaging up to 6 channels in multitrack configuration. The AxioObserver.Z1 inverted stand, is equipped with an environmental chamber with CO2 for live cell imaging. Focus stabilisation is through software and hardware based mechanisms (Definite focus II). Interchangeable stage inserts for slides, chamber slides, 35mm dishes and multiwall plates. Image capture in 3D and 4D, plus Lambda scanning, and spectral un-mixing capability.
lsm 880 with airyscan
Airyscanning
An additional scan detector providing 1.7x isotropic increase in resolution from widefield ~120nm. The Airy Disk or Point spread function from every light emitting point is projected through zoom optics and scanned by a hexagonal array for 32 GaAsP detectors which then undergoes a process of pixel reassignment and subsequent processing. All dyes and fluorophores that are scanned by the conventional detectors can be Airy-scanned using specific emission filter sets in simultaneous or sequential acquisition modes.
Airyscanning in Fast mode
In fast mode the beam is shaped to scan not one but 4 lines of pixels in each scanning sweep, acquiring 4x faster than normal Airyscan. Only the central 16 elements of the array are utilised providing a 1.5 x increase in resolution. Excellent for fast imaging of live cell dynamics with increased resolution
lsm 880 with airyscan
ZEISS Webinar: LSM 880 with Airyscan - Revolutionize Your Confocal Imaging
ZEISS LSM 880 with Airyscan: Beam Path and Principle of Airyscanning
Optics
| Objective lens | Magnification / NA / Immersion | Contrast Method |
|---|
|
C-ACHROMAT
|
10x / 0.45 / Water
|
|
|
LD LCI Plan-NEOFLUAR
|
25x / 0.8 / Water or Oil
|
DIC II
|
|
Plan-ACHROMAT
|
40x / 1.2 / Water
|
DIC III
|
|
Plan-APOCHROMAT
|
63x / 1.2 / Water
|
DIC III
|
|
Plan-APOCHROMAT
|
63x / 1.4 / Oil
|
DICIII
|
|
Plan-APOCHROMAT
|
100x / 1.4 / Oil
|
DIC III
|
Epifluorescence Filter Sets
| Filter set | Ex BP nm | Em BP nm |
|---|
|
Fs49 wf
|
365
|
445/50
|
|
Fs38 wf
|
470/40
|
525/50
|
|
Fs43 wf
|
545/25
|
635/70
|
Airy Scan Emission Filter Sets
|
BP420-445
|
+
|
BP465-505
|
|
BP420-480
|
+
|
BP495-550
|
|
BP495-550
|
+
|
LP525
|
|
BP495-535
|
+
|
LP555
|
|
BP495-550
|
+
|
LP570
|
| BP570-620 |
+ |
LP645 |
Laser Lines
- Lasos 30mW Diode 405nm
- Lasos 25mW LGN3001 multiline Argon, 458, 488, 514nm
- Lasos 20mW (DPSS 561) 561nm
- Lasos 2mW HeNe 594nm
- Lasos 5mW HeNe 633nm
Experimental Modes
- Z stack
- Multi-positions
- Time lapse
- Tile scanning / image stitching of large samples
- Bleaching and FRET analysis
Software and Network Drives
- Zen 2.3 allows for multidimensional experiment design in 2D, 3D, and 4D
- PC, HPZ840 4TB 3 RAID Hard drives and 64GB of memory.
- Desk top script connections to personal offline storage folders
Location
Imaging Suite, Room WXG86
Zeiss Lightsheet Z.1 Selective Plane Illumination Microscope
Unlike the Confocal microscope the Light Sheet microscope illuminates a single, thin plane at one time thus producing an optical section without the need for a confocal pinhole, but increasing SNR significantly. The detection lens is set at 90 degrees to the plane of illumination, and this perpendicular light path and focal plane illumination significantly reduces photo toxicity of delicate samples.
Application and advantages
Light sheet fluorescence microscopy is the way to observe small animal models, such as Zebrafish and Drosophila in a more gentle and physiological manner, but is also applicable to Arabidopsis, marine organisms, excised tissue and biopsies.
The sample is usually embedded in agarose and mounted vertically in an imaging chamber via glass a capillary. This allows the sample to be rotated in 360O for optimisation of the light sheet angle, but also to obtain a “Multiview” of a sample for whole organism reconstruction.
The system does not image pixel by pixel but will grab full frame using two sCMOS cameras allowing high resolution time lapse recordings for efficient analysis of dynamics within the sample.
Further background reading including downloads on sample preparation and other protocols
Specification
- Twin sCMOS PCO Edge 16bit cameras for simultaneous 2 channel acquisition
- Twin illumination objectives for single or dual sided sample illumination with dual side fusion of the live image
- Pivot scanning to reduce shadowing
- Time lapse
- Multiview
- Chamber incubation with CO2 available
- System configured for imaging cleared tissue
- Objective lenses, 10x 20x and 40x
- Laser lines, 405,445, 488,514,561 and 638nm
- 32TB 3 RAID drive processing workstation networked to RDS
Software and Network Drives
- Zen SP1 Lightsheet, allows for multidimensional experiment design in 3D, and 4D
- PC 1, HPZ840 4TB acquisition workstation
- PC 2, 32TB 3 RAID drive image processing workstation memory.
Location
Imaging Suite, Room WXG89
Zeiss Axio Scan .Z1 Slide Scanner
Axio Scan .Z1
The fully automated Zeiss Axio Scan .Z1 slide scanner creates high resolution virtual slides of whole tissue sections. Full tissue scanning with automatic or interactive detection of the selected area. The large capacity 100 slide magazine allows for scanning of large batches. Easy slide loading with dedicated imaging frames. Fully adaptable and programmable with synchronised external high speed excitation and internal emission filter wheels. Auto file saving to personalised storage in a secure server location.
ZEISS Axio Scan.Z1 - Your Virtual Slide Scanner for High Throughput Whole Slide Imaging
Sample illumination and cameras
| Light Source | Camera | Pixel size and resolution | Function |
|---|
|
Bright field
LED
|
3CCD colour 2MP Hitachi 1200x1600 HV-F202SCL
|
4.4 x 4.4 µm
0.22 µm per pixel*
|
Immunohistochemistry e.g. H&E, HRP, IHC
|
|
HXP120 fluorescent white light
|
Hamamatsu Orca Flash 4 -4MP 2048x204816bit sCMOS
|
6.45 x 6.45 µm 0.32 µm / pixel*
|
Multichannel fluorescence
|
|
|
|
*20x magnification
|
|
Optics
| Objective lens | Magnification / NA |
|---|
|
Fluar
|
5x / 0.25 M27 (course focusing)
|
|
Plan-Apochromatic
|
10x / 0.45 M27
|
|
Plan-Apochromatic
|
20x / 0.8 M27
|
|
Plan-Apochromatic
|
40x/ 0.95 M27
|
Fluorescence filter sets
| Filter set | Excitation | Beam Splitter | Emission | Examples of fluorophore combinations |
|---|
|
Zeiss 49 HE
|
365
|
FT385
|
445/50
|
Dapi / Hoescht / BV421/ Pacific Blue/ eFluor450 / Alexa 405 / Alexa 350
|
|
Zeiss 38 HE
|
470/40
|
FT495
|
525/40
|
eGFP / YFP / Alexa 488 / FITC / pacific Green / Qdot525 (with ET380nm exciter) / Horizon BV500 / BV510
|
|
Zeiss 43 HE
|
545/25
|
FT570HE
|
635/70
|
Cy3 / Alexa 555/ Rhodamine / pacific Orange / Qdot605 (with ET380nm exciter) / tdTomato / Texas Red / eFLuor605nv / BV605 / Propidium Iodide / PE
|
|
Zeiss 64 HE
|
587/25
|
FT605HE
|
647/70
|
mPlum / Alexa 594 / Alexa 610 / Texas Red / Qdot655 (with ET 380nm exciter) / BV650 /
|
|
Chroma ET49006
|
620/60
|
ET700/75
|
700/75
|
Cy5 / Alexa 633 / 647 / 700 / Qdot705 (with ET380nm exciter)
|
Slide acceptability and capacity
Single width slide; (max 4 per mounting frame) x 25 width 24-26mm, length 74-76mm, depth 0.8-1.3mm.
Double width slide; (max 2 per mounting frame) x 2 width 52mm, length 74-76mm, depth 0.8-1.3mm.
Quad width slide; (max 1 per frame) width 104mm, length 74-76mm, depth 0.8-1.3mm (optional extra).
Software
ZEN blue (2012) slide scan software has capability to drive whole tissue or multi-region tiling with Z-stacking plus specific profiles for TMA’s and cell spreads etc.
Location
Imaging Suite: Room WXG87
Leica DM6000B upright widefield epifluorescence and colour microscope
The Leica DM6000B is multimodal and equipped with twin Leica cameras for capture in 4 fluorescence channels and colour bright field for histological examination. Based on an upright DM6000B stand with fully motorised stage and filter turret. Applicable for slide analysis only.
Leica Microsystems
Optics
| Objective lens | Magnification / NA / Immersion | Contrast Method |
|---|
|
HC Plan Apo
|
10x / 0.45 / Dry
|
Phase 1
|
|
HC Plan Apo
|
20x / 0.7 / Dry
|
Phase 3
|
|
Plan Apo
|
40x / 1.25 / Oil
|
|
|
Plan Apo
|
100x / 1.4 / Oil
|
Phase 3
|
Epifluorescence filter sets
| Filter cube | Excitation nm | Emission nm | Fluorophores |
|---|
|
A4
|
360
|
Blue 470
|
Dapi / Hoescht / Pacific Blue / eFluor450 / Alexa Fluor 405
|
|
L5
|
480
|
Green 527
|
eGFP / YFP / Alexa 488 / FITC / pacific Green
|
|
N3
|
546
|
Red 600
|
Cy3 / Alexa 555/ Rhodamine / pacific Orange / tdTomato / Texas Red / Propidium Iodide / PE
|
|
Y5
|
620
|
Far Red 700
|
Cy5 / Alexa 633 / 647 / APC
|
Software and Network Drives
- Leica LASX core
- Desk top script connections to personal offline storage folders
Location
Confocal Microscopy Room, Centre for Translational Inflammation Research
Institute of Inflammation and Ageing, QEH/B
Leica DMI6000B widefield epifluorescence and brightfield microscope with timelapse
Based in the Centre for Translational Inflammation Research (CTIR), the Leica DMI 6000B is equipped with a DFL350 FX camera for capture in 3 fluorescence channels and bright field. Based on a fully motorised inverted DMI 6000B stand and environmental chamber with CO2 for live cell imaging. A time-lapse module is available for migration studies. Applicable for imaging slides, cell dishes, multi-well plates and chamber slides.
Leica Microsystems
Optics
| Objective lens | Magnification / NA / Immersion | Contrast Method |
|---|
|
HCX Pl Fluotar
|
20x / 0.4 / Corr Dry
|
Phase 1
|
|
HCX Pl Fluotar
|
40x / 0.6 / Corr Dry
|
Phase 2
|
|
HCX Pl Fluotar
|
63x / 0.7 / Corr Dry
|
Phase 2
|
| Filter cube | Excitation nm | Emissionnm | Fluorophores |
|---|
|
A4
|
360
|
Blue 470
|
Dapi / Hoescht / Pacific Blue / eFluor450 / Alexa Fluor 405
|
|
L5
|
480
|
Green 527
|
eGFP / YFP / Alexa 488 / FITC / pacific Green
|
|
N3
|
546
|
Red 600
|
Cy3 / Alexa 555/ Rhodamine / pacific Orange / tdTomato / Texas Red / Propidium Iodide / PE
|
Software and Network Drives
- Leica LASX core
- Desk top script connections to personal offline storage folders
Location
Confocal Microscopy Room, Centre for Translational Inflammation Research
Institute of Inflammation and Ageing, QEH/B
Zeiss PALM MicroBeam Laser Capture Microdissection
LCM Is the contamination-free method for separating and collecting cells of interest out of tissue samples mounted on slides or live cell cultures. The isolated pure cell populations are the perfect starting point for specific genetic, proteomic and molecular analysis. PALM MicroBeam makes isolating uncontaminated source material simple, via precise detection of specified areas, and contact free laser microdissection technology, ensuring your results are exact and reproducible. The PALM laser technology can also be used for DNA damage of live cells which can be coupled with live cell time lapse fluorescence imaging of the downstream effects.
Background and introduction to Zeiss PALM LCM
ZEISS Laser Capture Microdissection (LCM): Isolate cells by the force of light
ZEISS Laser Capture Microdissection (LCM): Bridging the Gap between LCM and Proteomics
Microscope stand
- Axio Observer.Z1 inverted microscope
- Environmental camber for live cell work
- Fully motorised stage with joystick
- Twin CCD cameras for colour brightfield (AxioCamCc1) and greyscale fluorescence (AxioCamMRM)
- Sample illumination through 100W HBO Mercury epi-fluorescence burner and HAL 100W tungsten transmitted light
- Sample collection to 200µl and 500µl PCR tubes or Zeiss adhesive caps.
Optics
| Objective lens | Magnification / NA / Immersion | Contrast Method |
|---|
|
Fluar
|
5x / 0.25
|
|
|
Fluar
|
10x / 0.5
|
|
|
LD Plan Neofluer
|
20x / 0.4 / Korr
|
|
|
LD Plan Neofluer
|
40x / 0.6/ Korr
|
|
|
EC Plan Neofluer
|
100x / 1.3 / Oil
|
|
|
Plan Apochromat
|
150x / 1.35 / Oil
|
DIC
|
Widefield Epi-fluorescence Dichroic Filter sets
| Filter set | Ex BPnm | Em BPnm | Fluorophores |
|---|
|
FS49
|
365
|
445/50
|
Dapi / Hoescht / Pacific Blue / eFluor450 / Alexa Fluor 405
|
|
FS38 HE
|
470/40
|
525/50
|
eGFP / YFP / Alexa 488 / FITC / pacific Green
|
|
FS20
|
546/12
|
575/640
|
Cy3 / Alexa 555/ Rhodamine / pacific Orange / tdTomato / Texas Red / Propidium Iodide / PE
|
Software and Network Drives
- Zeiss PALM Robo software version 4.6
- Desk top script connections to personal offline storage folders
Location
Imaging Suite: Room WXG87
Zeiss Axio Observer widefield system with Apotome 2.0 system
Motorised Axio Observer geared toward fluorescence performance with LED Colibri.2 illumination system that offers high speed switching times to the order of less than 1 ms. The unique feature of the Colibri.2 design in comparison with all other LED systems currently available is its unmatched homogeneity. The light path up to the illuminator output is the same for each LED module independent of its installation position. Four channels are included at 365nm, 470nm, 565nm, and 590nm. The system is equipped with an Hamamatsu ORCA Flash 4.0 for high speed and high sensitivity.
In addition, the system is equipped with the Apotome.2 Confocal Structured Illumination device for creating optical sections with high contrast. Despite the magnification used, the ApoTome.2 automatically places the right grid in the beam path of the microscope. Reduction of unwanted background fluorescence increases with the grid frequency and the optical sections become thinner. Structures from outside of the focal plane are suppressed improving contrast and resolution of the optical section.