Picoquant Microtime200

 

Fluorescence Lifetime Imaging (FLIM) adds another dimension to conventional intensity based imaging methods. FLIM is not affected by fluctuations in the fluorescence intensity and permits to discrimination between fluorophores with similar emission spectra (and from auto-fluorescence. It can be used to probe local environmental conditions (e.g., pH value), to determine ion concentrations, to study intracellular signal transduction or to distinguish between different tissue components.

 
icroscope body:
Inverted
Motroised Stage:
No
Temp Control:
Stage insert
CO2:
No
Software:
SymPhoTime 64

 

Common Applications

FLIM: Each fluorescent dye displays a characteristic fluorescence lifetime, namely the length of time following excitation that the dye will remain in its excited state emitting fluorescence.  This fluorescence lifetime is a quantitative signature that can be used to distinguish between different fluorescent dyes or spectrally similar fluorophores, even those emitting the same fluorescent colour. This technique can help determine local environment parameters (e.g., pH), analyze oxygen, water, or ion concentrations as well as differentiate structures deep inside tissue and eliminate background fluorescence
FCS: Fluorescence Correlating Spectroscopy allows for determining diffusion coefficients and concentrations of fluorescently labelled molecules at nanomolar concentrations, both in vitro and in live cells. It is mostly useful for indirect studies of molecular activity in plasma membrane, in cytosol and in nucleus via following relative changes in diffusive behavior and/or concentration. This technique can also determine the mobility of molecules (e.g. movement, aggregation, association, dissociation, and conformational changes), help studying the intracellular dynamics (e.g. diffusion, active transport) and determine the stoichiometry of complexes
FRET:

FRET (Lasers Förster Resonance Energy Transfer or Fluorescence Resonance Energy Transfer) is a technique using two carefully selected fluorophores that when in close enough proximity (10–100 Å), and suitable orientation, energy is transferred nonradiatively from the excited donor fluorophore to another acceptor fluorophore. It is largely used to investigate inter- and intramolecular distances, between one or more molecules. It can also monitor conformational changes of molecules and environmental parameters (e.g., pH) via FRET sensors.

 

Objective Lenses

MAG N.A CORRECTIONS IMMERSION W.D MISC.
20x 0.75 UPlanSApo Air ? ?
60z 1.2 UPlanSApo Water ? ?
100x 1.45 PlanApo Oil ? ?

 

Light Sources

Lasers Wavelengths
Diode 450 405nm
Diode 470 470nm
Diode 532 532nm
Diode 640 640nm

 

Filters

Detectors

Detector Timing Resolution Efficiency Description
2x  Single-Photon Avalanche Diodes - PDM series Down to < 50 ps (FWHM( Up to 49% at 550nm Solid-state photodector in which a photon-generated carrier can trigger a short-duration but relatively large avalanche currnt. Ultra stable at high count rates