<i>In vivo</i> laser speckle contrast imaging of microvascular blood perfusion using a chip-on-tip camera

Laser speckle contrast imaging (LSCI) is an important non-invasive capability for real-time imaging for tissue-perfusion assessment. Yet, the size and weight of current clinical standard LSCI instrumentation restricts usage to mainly peripheral skin perfusion. Miniaturization of LSCI could enable hand-held instrumentation to image internal organ/tissue to produce accurate speckle-perfusion maps. We characterized a 1mm 2 chip-on-tip camera for LSCI of blood perfusion in vivo and with a flow model. A dedicated optical setup was built to compare chip-on-tip camera to a high specification reference camera (GS3) for LSCI. We compared LSCI performance using a calibration standard and a flow phantom. Subsequently the camera assessed placenta perfusion in a small animal model. Lastly, a human study was conducted on the perfusion in fingertips of 13-volunteers. We demonstrate that the chip-on-tip camera can perform wide-field, in vivo, LSCI of tissue perfusion with the ability to measure physiological blood flow changes comparable with a standard reference camera.</p

New high-speed centre of mass method incorporating background subtraction for accurate determination of fluorescence lifetime

We demonstrate an implementation of a centre-of-mass method (CMM) incorporating background subtraction for use in multifocal fluorescence lifetime imaging microscopy to accurately determine fluorescence lifetime in live cell imaging using the Megaframe camera. The inclusion of background subtraction solves one of the major issues associated with centre-of-mass approaches, namely the sensitivity of the algorithm to background signal. The algorithm, which is predominantly implemented in hardware, provides real-time lifetime output and allows the user to effectively condense large amounts of photon data. Instead of requiring the transfer of thousands of photon arrival times, the lifetime is simply represented by one value which allows the system to collect data up to limit of pulse pile-up without any limitations on data transfer rates. In order to evaluate the performance of this new CMM algorithm with existing techniques (i.e. Rapid lifetime determination and Levenburg-Marquardt), we imaged live MCF-7 human breast carcinoma cells transiently transfected with FRET standards. We show that, it offers significant advantages in terms of lifetime accuracy and insensitivity to variability in dark count rate (DCR) between Megaframe camera pixels. Unlike other algorithms no prior knowledge of the expected lifetime is required to perform lifetime determination. The ability of this technique to provide real-time lifetime readout makes it extremely useful for a number of applications

Tissue surface as the reference arm in Fourier domain optical coherence tomography

We present a simple method applicable to common-path Fourier domain optical coherence tomography (OCT) in which the tissue surface is used as the reference arm. We propose using aluminium hydroxide powder as a potential tissue surface diffuser to allow wider application of this method. This technique allows one to avoid placing a reference arm reflective element, such as glass plate, on tissue, and intrinsically avoids both coherent and complex conjugate mirror artifacts associated with glass plates. Aluminium hydroxide can be sprayed onto tissue using spray nozzles commonly found in endoscopes. The sensitivity of the tissue reference arm common-path OCT image is 94 dB for a 50-mu s charge-coupled device integration time, and 97.5 dB for a 200-mu s CCD integration time. (C) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOL 10.1117/1.JBO.17.7.071305]Publisher PDFPeer reviewe