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pco.flim

pco.flim Camera System

Frequency domain fluorescence lifetime imaging camera.

 

Description

The pco.flim is the first Frequency Domain FLIM (Fluorescence Lifetime Imaging) camera system using a two tap CMOS sensor. Synchronized modulation of pixels and stimulated light enables you the analysis of luminescence decay times in the range of 100 ps to 100 µs. With its 1008 x 1008 pixels resolution the pco.flim reads out 45 double frames/s at a max. You can use it for a modulation frequency range of 5 kHz to 40 MHz.

Using C-mount as standard the system is easy to connect to any microscope or lens. Further, the USB 3.0 interface lets you connect the camera to all kinds of computers. This saves you significant efforts and costs for operation and research.

The pco.flim laser is a homogeneously illuminating light source designed for use with the pco.flim. It features a digital modulation frequency range of 0 to 250 MHz. You can either order the pco.flim laser for widefield epifluorescence microscope illumination, or for Total-Internal-Reflection-Fluorescence (TIRF) microscopy, light sheet fluorescence microscopy (LSFM) and confocal spinning disk microscopy.

Choose between a wide range of different wavelengths and light output powers. The laser is ideally suited if your relevant luminescence lifetimes are in the range of a few nanoseconds. In case you are using a Nikon microscope, the Nikon double-safety-shutter option is also available.

Key Features:

  • pco.flim laser as light source for epifluorescence measurements
  • selectable laser wavelengths and light output powers
  • 100 ps – 100 µs lifetimes measurable
  • 5 kHz – 40 MHz modulation frequencies
  • 500 kHz – 40 MHz external modulation signals
  • modulation signal shape sinusoidal / rectangular
  • 1008 x 1008 pixel resolution
  • frequency domain FLIM
  • USB 3.0 interface
  • 39 % quantum efficiency
  • 1000:1 dynamic range
  • 45 double frames/s (2 tap readout)
  • 1 ms to 2 s selectable exposure times
  • vibration-free water cooling
  • special software for measurement and analysis

Technical Specification:

 Unit pco.flim
Resolution (hor x vert)pixel1008 x 1008
Pixel size (hor x vert)µm5.6 x 5.6
Modulation frequency (out)5 kHz - 40 MHz
Modulation frequency (in)500 kHz - 40 MHz
Peak Quantum Efficiency%39
Dynamic Rangebit14

Quantum Efficiency Curve:

 

View/ Download the datasheet here.

Application Examples:

Widefield FLIM of Protoporphyrin IX with pco.flim

Widefield FLIM of Protoporphyrin IXAchieving a maximal safe extent of resection during brain tumor surgery is the goal for improved patient prognosis. Fluorescence-guided neurosurgery using 5-aminolevulinic acid (5-ALA) induced Protoporphyrin IX has thereby become a valuable tool enabling a high frequency of complete resections and a prolonged progression-free survival in glioblastoma patients.  A widefield fluorescence lifetime imaging device with 250 mm working distance working under similar conditions like surgical microscopes based on a time-of-flight based dual tap CMOS camera is presented. Read more about this here.

 

Fluorescence Lifetime Based pH MEAS 

Widefield FLIM of Protoporphyrin IXNovel fluorescent diazaoxatriangulenium (DAOTA) pH indicators for lifetime-based self-referenced pH sensing are reported. The DAOTA dyes were decorated with phenolic receptor groups inducing fluorescence quenching via photoinduced electron transfer mechanism. Read more about this here.

 

Luminescence Lifetime Imaging of Chemical Sensors with pco.flim

Widefield FLIM of Protoporphyrin IXLuminescence lifetime based imaging is still the most reliable method for generating chemical images using chemical sensor technology. However, only few commercial systems are available that enable imaging lifetimes within the relevant nanosecond to microsecond range. In this technical note we compare the performance of an older time-domain (TD) based camera system with a frequency-domain (FD) based camera system regarding their measuring characteristics and applicability for O2 and pH imaging in environmental samples and with different indicator dye systems emitting in the visible and near-infrared part of the spectrum. We conclude that the newly introduced FD imaging system delivers comparable if not better results than its predecessor, now enabling robust and simple chemical imaging based on FD luminescence lifetime measurements. Read more about this here.

 

Frequency Domain FLIM with pco.flim

Widefield FLIM of Protoporphyrin IXWidefield frequency-domain fluorescence lifetime imaging microscopy (FDFLIM) is a fast and accurate method to measure the fluorescence lifetime of entire images. However, the complexity and high costs involved in construction of such a system limit the extensive use of this technique. PCO AG recently released the first luminescence lifetime imaging camera based on a high frequency modulated CMOS image sensor, QMFLIM2. Read more about this here.

Fluorescence Lifetime Imaging (FLIM) Tutorial:

View / Download the whitepaper here.

For hands-on session from PCO’s Head of Research, Dr Gerhard Holst, see below:

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