EMCCDs

Figure 5. (A) Scheme of two-photon laser scanning microscope (1) Ti Sa laser, 100 fs, 80 MHz, 750-980 nm, 1.6W 800 nm (TSUNAMI, Spectra Physics), (2) pre-chirp, (3) beam multiplexer, (4) scanning mirrors, (5) microscope (Olympus IX 71, XLUMPLFL20XW, WD = 2 mm, NA = 0.95), (6) fluorescent foci in sample, (7) filter wheel/spectrograph (SpectraPro 2300i, Acton Research Corporation)/spectrometer (home built), (8) back illuminated EMCCD camera (IXON BV887ECS-UVB, Andor Technology), (9) dichroic mirror (2P-Beamsplitter 680 DCSPXR, Chroma). (B) Experimental setup of two-photon laser scanning microscope.

Figure 1.15 Signal-to-noise ratio of the CH2 symmetric stretching band of PMMA plotted against the gain of the EMCCD. Reproduced with permission from Ref [38].

Inverted microscope with EMCCD camera, equipped for TIRF or spinning-disc microscopy. 

Back illuminated, frame transfer electron-multiplying charge coupled device (EMCCD) camera (such as Photometries Cascade II 512, Roper Scientific, Tuscon, AZ). Back illuminated EMCCD cameras capture more than 90% of incoming photons and amplify signals to overcome instrument noise. 

Set the frame rate of the image acquisition to 10-200 Hz. To achieve the fast range of frame rates (i.e., >30 Hz), the active imaging region of the EMCCD camera usually needs to be reduced. 

Due to the particle nature of photons, it is unlikely that each pixel on the EMCCD camera receives the exact same number of photons per time. Consequently, the value of each pixel, which contributes to a single-molecule signal, will vary over time. 

ICCD/EMCCD Stoichiometry determination [62] GFP Imaging In cells [9] Nucleotide kinetics [67] Combined TIRF/AFM [72]... 

FRET fluorescence resonance energy transfer FCS fluorescence correlation spectroscopy TIRF total internal reflection fiuorescence PCFI photon counting histogram ICCD intensified charge coupled device EMCCD eiectron muitipiying charge coupled device CMOS complimentary metal oxide semiconductor AFM atomic force microscope. 

Figure 3.21 Schematic representation of the structure of a typical CCD. Elements are read out sequentially by first moving charge downwards line by into an output register using a series of electrodes. This register is then read out one element at a time and the signals amplified by an external electronic current amplifier. In the EMCCD systems on-chip gain is provided before external amplification to lift even very low signals well above...

CCD Camera Back illuminated EMCCD (Andor iXon,Andor Technology, Northern Ireland)... 

Figure 14 Diagram depicting the experimental setup for the upconversion luminescence in vivo imaging system designed by Li s group. Two external 0-5 W adjustable CW 980 nm lasers were used as the excitation sources, and an Andor DU897 EMCCD was used as the signal collector. (Reproduced with permission from Ref. 72. Copyright (2009) American Chemical Society.)...

Fig. 24.8 Schematic of the exptaimcmtal setup (M minor, BS beam splitter, SL spherical lens, CL cylindrical lens, BD beam dump, DM dichroic mirror, NDF neutral density filter, BPF bandpass filtea, LPF longpass filto, EMCCD-C electron multiplying charge-coupled device camera) (from [24])...

EMCCD camera with high quantum efficiency (-90 % at 670 nm) and capable of achieving frame rates in the order of 30-50 frames per second. 

---