Ionic imaging: inverted microscope
This setup allows recordings of ionic fluxes such as calcium fluxes in cells loaded with fluorophores (Fura-2, Mag-Fura-2, Indo-1, ...) and monitoring of ionic currents using the Patch-clamp technique. Cells can also be electrically stimulated during recordings.
Different substances such as drugs can be applied on cells using the perfusion system. The recording chamber can be heated to work in conditions close to the physiological temperature.
System features
Microscope : Olympus IX71, inverted, non motorised
DIC condenser
Light sources :
Monochromator Polychrome V (TILL Photonics)
Halogen lamp for transmission + Uniblitz shutter
Objectives: Olympus
- Plan Apochromat 10 X, numerical aperture 0.40
- Plan semi Apochromat 40 X long working distance, numerical aperture 0.60
- UV Apochromat 40 X , oil immersion, numerical aperture 1.35, special for UV excitation at 340 nm
- Plan semi Apochromat 60 X, oil immersion, numerical aperture 1.25
- Optovar lens 1.6x
Filters in the filter wheel:
- FITC : HQ480/40x, Q505LP, HQ535/50m
- Fura-2 : no excitation filter, 400DCLP, D510/80m
- Indo-1 : D365/10x , 380DCLP
Filters in the Dual View (for simultaneous acquisitions):
- Fura 2 / IR visible: D520/60m, 565ldx, D660p/10m
- Indo1 (2 emissions): D405/30m, 440dclp, D485/25m
Camera :
CCD CoolSnap HQ monochrome (Photometrics)
Softwares:
Metamorph and Metafluor (Molecular Devices)
Axon pClamp 10
Electrophysiology:
Micro-manipulator Exfo Burleigh
Axopatch 200B
Digidata 1440
Electrical stimulator électrique HAMEG
Accessories:
Objectve heater (40x oil)
Imaging chamber heater
Perfusion system
Perfusion chambers for rounded coverslips, diameters: 15, 18 and 25 mm