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Single Photon Counting Imager


Single photon counting camera
LINCam is a solution for scanning-free time correlated single photon counting implemented as a camera. This camera resolves x and y positions of individual photons as precise as a CCD with 1000 × 1000 pixels does together with 50 ps accuracy timing. Being paired with a pulsed light source LINCam turns any conventional fluorescence microscope into a powerful lifetime measuring instrument. LINCam with attached off-the-shelf optics is a solution for macroscopic applications like LIDAR.

In other words, LINCam is just a camera. As easy as an ordinal megapixel CCD camera but extended with the timing dimension.


Acquisition system
Universal electronics and software for LINCam25 and LINCam40. The system grants single photon acquisition in a wide-field counting mode. The core of the system is a position sensitive photomultiplier tube (PMT) based on microchannel plates (MPC’s) with a multi-alkali photocathode. The system comprises of the detector head and the electronic control module designed to be used in lab conditions. The detector head houses MCP-PMT preamplifiers, high-voltage power suppliers and the cooling system. Provided integrated electronic control module includes everything required for robust and reliable single photon counting based imaging. Real-time event selection logic processes registered photons to avoid artifacts like multi-photon events, MCP noise and pile-up effect.


Application

• Fluorescence lifetime imaging (FLIM)

• Light-sheet 3D FLIM
• Time resolved Raman spectroscopy
• Time-of-Flight measurements

• Low-light observations

QE_R.JPG



Product specifications and Brochures

Product Brochure Link: 


Detector 
LINCam25 
LINCam40
Active Area Diameter, mm
2540
Positional Resolution, pixels1000 x 1000
Temporal Resolution, ps FWHM≤50
Microscope MountC-mountT-mount
Housing Dimentions, mm145 x 78 x 50140 x 100 x53
Weight, g500600
CoolingLow Noise Air or Liquid


 Acquisition system

 Maximal Count Rate, MHz

1
 Dead Time, ns400
 Timing
 Method
TAC+ADC
 Minimum Bin Width, ps≤1,4
 Electrical Resolution, ps6
 Number of Bins4096
 Reference InputPositive or negative NIM
 Time Tagging Resolution, ns

10

 Computer InterfaceUSB 2.0
 Operating SystemWindows 7/10 64 Bit



Confocal Superresolution FLIM Microscopy





Photoscore LINcam used here as a drop-in replacement of the hamamatsu CCD camera. The detector housing has a c-mount female thread that allows a direct attachment to the CSU. Acquisition electronic module controls the camera and transfers acquired data to the computer with a standard USB interface. A pulsed laser is required to perform time-resolved single photon counting. Here, Omicron QuixX diode laser was coupled into a light input of CSU using a single mode fibre.


Primary hippocampal neurons from rats.To visualize excitatory synaptic contacts, 

neurons were stained with rat anti-homer, guinea pig anti-MAP2, rat anti-Ctip2 

and mouse anti- Prox1 antibodies. Subsequently, samples were incubated with 

anti-rat Alexa 488-, anti-guinea pig Cy5-, and anti-mouse Alexa 350- conjugated 

donkey secondary antibodies.

               




       

  

Lymphocytes (Jurakt T-cells) were transfected with a monomeric CFP-YFP Lck-biosensor and stimulated by CD3. After fixation cells were immune stained by an anti-GFP antibody, labelled with Atto 647N. The optical sections clearly show the shuttling of Lck positive vesicle between the plasma membrane and an inner compartment (most likely associated with the Golgi complex). Intensity weighted 3D stack of FLIM images of T-cells, 400 × 400 bins, 20 seconds per slice.



Light-sheet Lifetime Imaging

Rat embryo acquired with light-sheet microscope


One of the well-known drawbacks of widefield microscopy imaging its 

low axial resolution compared to confocal microscopy. Sevaral methods are 

known to overcome those limitations introducing confocality. Here we 

demonstrate FLIM images acquired with optical sectioning with light-sheet illumination.


Drosophila larvae


A commercially available light-sheet system equipped with a pulsed laser source and a CCD with c-mount. For the user it is just a drop-in replacement of the CCD by LINCam to start imaging. An image below shows 128 FLIM sections acquired within 10 seconds per frame.


FILM (Fluorescence lifetime Imaging)

Fluorescence Lifetime Imaging (FLIM) is a technique which uses the separation of different fluorescence decay times of fluorophores to create an image contrast other than intensity in classical imaging.

Example of lifetime imaging of a lily of the valley slice sample. The intensity image 

(a) is a histogram of the positions of acquired photons. Lifetime analysis reveals four 

lifetime components: τ1 = 0,19; τ2 = 0,67; τ3 = 1,95 and τ4 = 3,75 ns. The resulting 

overlay image 

(b) of the intensity image and average lifetime is shown.


Glycolytic Oscillations in Eukaryotic Cells Followed by NADH Imaging

By using the metabolite NADH as an intrinsic marker for glycolysis, the dynamics of individual cells can be monitored and their interactions studied.

Monitoring intrinsic energy metabolism over long periods of time allows the study of cellular communication between cell populations. By using the metabolite NADH as an intrinsic marker for glycolysis, the dynamics of individual cells can be monitored and their interactions studied. Glucose consumption by glycolysis and alcoholic fermentation leads to the production of metabolites, some of which are released. Coupling between yeast cells depends on the release and sensing of the messenger acetaldehyde, which diffuses through the extracellular medium. Yeast cells are well known for the oscillatory behavior of the glycolysis and their metabolic organization. The exchange of messenger molecules can result in waves and synchronized patterns in which all cells oscillate in concert. Essential to this study is an ultrasensitive detection system that allows excitation of the weak fluorescence of NADH by low-intensity UV light.



(a) The time-series of the collective NADH fluorescence signal for a yeast population of cell density ρ=0.1%. Partial synchronisation of intracellular oscillations occurs at 

760 s ≤t≤1100 s. (b) Development of the relative amplitudes of oscillations of each cell, and (c) of their phases. In (b) and (c) the cells are sorted according to their phases at time t=900 s. (d) Evolution of the distribution of instantaneous frequencies fi of the cells, and (e) of the distribution of the phase difference Δϕi between the phase ϕi of each individual cell to that of the average phase Φ of all cells of the population. (f) Time dependence of the order parameter R. The field of view had a diameter of 169 μm, and hosted 232 cells. Glucose was added to the cell suspension at t=−158 s.





Measurement induced entanglement of stored ions

Typically single photon emitters show an emission behaviour that is called anti-bunching. This means that the probability to detect a second photon after detection a first one is suppressed. By using two of this single photon emitters in form of trapped calcium ions, it is possible to create a ligh source consisting of two single calcium ions.


If the scattered light does not contain information of its creating ion (achieved by measuring the far-field), some interesting properties of this light source can be observed. As described in fig. 1, the two ion system can be described by the Dicke-Basis (Wolf et al. 2020)*. Interestingly the anti symmetric state |a> does not couple to the laser field. The system now shows behaviour of typical single photon emitters, an anti-bunched photon statistics, if it driven in the symmetric decay channel. If a photon is emitted in a way that the system ends up in the anti-symmetric case, it becomes invisible for the driving laser and a second photon has to be emitted to bring the system back t0 the ground state. Such behaviour of the emission of two photons in a short period of time is called bunching and can be found normally only for chaotic or thermal light sources.


Interestingly the decay channel can be chosen by the angle of investigation of the trapped ion crystal, deliviring a light source whose emission statistics can be tuned from non-classical anti-bunching to classical bunching and everything in between by just changing the angle under which it is observed.

In fig. 2 the used setup is illustrated. Two calcium ions are stored in a segmented Paul trap and cooled to the Doppler-Limit. The emitted fluorescence light is collected by a lense that images the ion crystal on a amplified CCD camera. Only 10% of the light is used for this, the other 90% of the light are reflected by a 90/10 beam splitter to a Hanbury-Brown and Twiss (HBT) setup in which the detectors are replaced by two spatially resolving LINCam systems. The CCD camera is used to monitor the status of the ion crystal, so that the measurement can be paused upon ion loss and restarted once the crystals is ready again. The HBT setup is used to record all incomingdouble photon events with a coincidence window of +-40ns and a precision of 50ps at 1000*1000 spatial pixels.

From this event stream, the second order auto and cross-correlation function can be calculated on the fly. The resulting histogram is displayed live to monitor the success of the experiment.


In [1] the measurement was conducted by using avalanche photo diodes and a TDC (Time-To-Digital-Converter). The angle of observation was chosen by a slit. With this method the majority of the light is discarded and thus the measurement takes 2-3 days per point to gather sufficient statistics. In fig 3. the results of this measurement campaign can be seen. The acquisition of the 8 measured angles took roughly 30 days.

In [2] and fig.4 the LINCam was used to redo the experiment with the aim of measuring more than 8 angles. By using two synchronized LINCams it was possible to record a two-photon-event stream and correlating those events on the fly. The measurement campaign took 30 days again but delivered not only 8 observation angles, but 96. With the APD setup the campaing would have last one year.




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Compare Model Drawings & Specs Availability Reference Price
(USD)
LINCAM25
Single photon counting camera, Active area diameter 25 nm, Positional resolution 1000 X 1000 px, Temporal resolution ≤50 ps, Microscopy Mount C-mount
4-6 Weeks Request for quote
LINCam40
Single photon counting camera, Active area diameter 40 nm, Positional resolution 1000 X 1000 px, Temporal resolution ≤50 ps, Microscopy Mount T-mount
4-6 Weeks Request for quote

LINCam40 - Parameter

LINCAM25 - Parameter

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LINCAM25 - Download

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