Fluorescence Microscopes

2 fluorescent inverted microscopes are available:

Zeiss Axiovert 200

Equipped with:

Environmental chamber
Eppendorf Micromanipulators
Eppendorf Microinjector
Piezo focus drive
Polychromator
Hamamatsu C4880-80 B/W digital fluorescent camera
Jenoptics colour digital camera (fluorescence capable)

Filter sets:

FITC, Texas Red, and DAPI
FURA-2

Software:

Improvision Acquisition and Analysis software

Functions:

Ca2+ ratioing
Fluorescence
Optical sectioning
Task automation
Time Lapse
Phase Contrast

Zeiss Axiovert 100M

Equipped with:

Polychromator
Hamamatsu C4742-95 B/W digital fluorescence camera

Filter sets:

FITC, Cy5, DAPI, TRITC, Cy3
CMXRos, JC-1

Software:

Improvision Acquisition and Analysis software

Functions:

Fluorescence
Optical sectioning
Task automation
Time lapse
Phase contrast


2 fluorescent upright microscopes are available:

Zeiss Axioskop

Equipped with:

Hamamatsu C4742-95 B/W digital fluorescence camera


Filter Sets:

FITC, TRITC, DAPI

Software:

Improvision Acquisition and Analysis software

Functions:

Fluorescence
Task automation
Time lapse

Zeiss Axioskop II

Equipped with:

Hamamatsu Orca-ER B/W digital fluorescence camera.


Filter Sets:

FITC, Texas Red, DAPI

Software:

Improvision Acquisition and Analysis software

Functions:

Fluorescence
Optical Sectioning
Task Automation
Time Lapse
Phase Contrast


Fluorescence Gallery - click to launch a full-size version.

Facilities Pulmonary

Pulmonary artery smooth muscle cells labelled for monomeric G-actin (green) and F-actin (red).

Facilities Cultured Aortic


Cultured aortic endothelial cells transfected with FAK protein (green). Cells are labelled for VE-cadherin (red) and cell nuclei are labelled blue.

Facilities Cultured Aortic Endothelial


Cultured aortic endothelial cells. Green labelling denotes phosphorylated FAK at the red labelled cellular junctions. The cell nuclei are labelled blue

Facilities Cultured HEK

Cultured HEK cells transfected with GFP, nuclei are labelled blue

Confocal Microcope The confocal microscope is a Bio-Rad Radiance 2000. The following lines are available:

Laser Wavelength Fluorophore
Argon 457nm
476nm
488nm
514nm 		CFP
GFP
FITC
Cy3
Greeen HeNe 543nm Texas red
Red Diode

637nm Cy5

(Note. Fluorophores with similar excitation/emission spectra will work)

The microscope scanhead is swappable between a Nikon TE300 inverted or a Nikon E1000 Upright microscope

Supporting equipment for the confocal:

Burleigh Micromanipulators
Perfusion System
Heated Stage 

Cardiac myocytes labelled with TMRM showing activated (right) and inactivated (left) mitochondria.

Image Analysis

There are 3 image analysis packages available witin the suite

  1. Improvision Openlab 3.1

    Analysis Modules:
    Morphometry
    Task automation
    Density slicing
    Nearest neighbour image deconvolution
    Volume deconvolution
    3D deconvolution
    3D reconstruction

  2. LaserPix

    Analysis Modules:
    Morphometry
    Co-localisation
    Bleed through correction
    Time course analysis
    Relative quantitation
    Population density
    Report generator

  3. LaserVox

    Analysis Modules:
    3D Morphometry
    3D Rendering


Booking Information and Costs

The non-confocal microscopes are booked on a half day basis

Morning: 9am-1pm
Afternoon: 1pm-5pm

The confocal microscopes are booked on a 3.5hour session basis.

08:00-11:30
11:30-15:00
15:00-18:30

Confocal Charges:
British Heart Foundation Staff £15 per session.
Rayne Institute Staff £30 per session.
Other £30 per hour (minimum 2 hours)

All users must be trained by the imaging suite manager prior to use, regardless of experience.