Optical Mapping of Cardiac Stimulation: Fluorescent Imaging with a Photodiode Array

2002 ◽  
pp. 596-635
2021 ◽  
Author(s):  
Dmitry Rybashlykov ◽  
Jaclyn Brennan ◽  
Zexu Lin ◽  
Igor R. Efimov ◽  
Roman Syunyaev

Fluorescent imaging with voltage- or calcium-sensitive dyes, i.e. optical mapping, is one of the indispensable modern techniques to study cardiac electrophysiology, unsurpassed by temporal and spatial resolution. High-speed CMOS cameras capable of optical registration of action potential propagation are in general very costly. We present a complete solution priced below US$1,000 (including camera and lens) at the moment of publication with an open-source image acquisition and processing software. We demonstrate that the iDS UI-3130CP rev.2 camera we used in this study is capable of 200x200 977 frames per second (FPS) action potential recordings from rodent hearts. The signal-to-noise-ratio of a conditioned signal was 16 ± 10 for rodent hearts. A comparison with a specialized MiCAM Ultimate-L camera has shown that signal-to-noise ratio (SNR) is sufficient for accurate measurements of AP waveform, conduction velocity (± 0.04 m/s) and action potential duration (± 7ms) in mouse and rat hearts. We measured the action potential prolongation during 4-aminopyridine administration in mouse heart, showing that proposed system signal quality is adequate for drug studies.


Author(s):  
J. B. Warren

Electron diffraction intensity profiles have been used extensively in studies of polycrystalline and amorphous thin films. In previous work, diffraction intensity profiles were quantitized either by mechanically scanning the photographic emulsion with a densitometer or by using deflection coils to scan the diffraction pattern over a stationary detector. Such methods tend to be slow, and the intensities must still be converted from analog to digital form for quantitative analysis. The Instrumentation Division at Brookhaven has designed and constructed a electron diffractometer, based on a silicon photodiode array, that overcomes these disadvantages. The instrument is compact (Fig. 1), can be used with any unmodified electron microscope, and acquires the data in a form immediately accessible by microcomputer.Major components include a RETICON 1024 element photodiode array for the de tector, an Analog Devices MAS-1202 analog digital converter and a Digital Equipment LSI 11/2 microcomputer. The photodiode array cannot detect high energy electrons without damage so an f/1.4 lens is used to focus the phosphor screen image of the diffraction pattern on to the photodiode array.


Author(s):  
J. Barbillat ◽  
M. Delhaye ◽  
P. Dhamelincourt

Raman mapping, with a spatial resolution close to the diffraction limit, can help to reveal the distribution of chemical species at the surface of an heterogeneous sample.As early as 1975,three methods of sample laser illumination and detector configuration have been proposed to perform Raman mapping at the microscopic level (Fig. 1),:- Point illumination:The basic design of the instrument is a classical Raman microprobe equipped with a PM tube or either a linear photodiode array or a two-dimensional CCD detector. A laser beam is focused on a very small area ,close to the diffraction limit.In order to explore the whole surface of the sample,the specimen is moved sequentially beneath the microscope by means of a motorized XY stage. For each point analyzed, a complete spectrum is obtained from which spectral information of interest is extracted for Raman image reconstruction.- Line illuminationA narrow laser line is focused onto the sample either by a cylindrical lens or by a scanning device and is optically conjugated with the entrance slit of the stigmatic spectrograph.


2020 ◽  
Vol 48 (6) ◽  
pp. 2657-2667
Author(s):  
Felipe Montecinos-Franjola ◽  
John Y. Lin ◽  
Erik A. Rodriguez

Noninvasive fluorescent imaging requires far-red and near-infrared fluorescent proteins for deeper imaging. Near-infrared light penetrates biological tissue with blood vessels due to low absorbance, scattering, and reflection of light and has a greater signal-to-noise due to less autofluorescence. Far-red and near-infrared fluorescent proteins absorb light >600 nm to expand the color palette for imaging multiple biosensors and noninvasive in vivo imaging. The ideal fluorescent proteins are bright, photobleach minimally, express well in the desired cells, do not oligomerize, and generate or incorporate exogenous fluorophores efficiently. Coral-derived red fluorescent proteins require oxygen for fluorophore formation and release two hydrogen peroxide molecules. New fluorescent proteins based on phytochrome and phycobiliproteins use biliverdin IXα as fluorophores, do not require oxygen for maturation to image anaerobic organisms and tumor core, and do not generate hydrogen peroxide. The small Ultra-Red Fluorescent Protein (smURFP) was evolved from a cyanobacterial phycobiliprotein to covalently attach biliverdin as an exogenous fluorophore. The small Ultra-Red Fluorescent Protein is biophysically as bright as the enhanced green fluorescent protein, is exceptionally photostable, used for biosensor development, and visible in living mice. Novel applications of smURFP include in vitro protein diagnostics with attomolar (10−18 M) sensitivity, encapsulation in viral particles, and fluorescent protein nanoparticles. However, the availability of biliverdin limits the fluorescence of biliverdin-attaching fluorescent proteins; hence, extra biliverdin is needed to enhance brightness. New methods for improved biliverdin bioavailability are necessary to develop improved bright far-red and near-infrared fluorescent proteins for noninvasive imaging in vivo.


VASA ◽  
2017 ◽  
Vol 46 (5) ◽  
pp. 383-388 ◽  
Author(s):  
Henrik Christian Rieß ◽  
Anna Duprée ◽  
Christian-Alexander Behrendt ◽  
Tilo Kölbel ◽  
Eike Sebastian Debus ◽  
...  

Abstract. Background: Perioperative evaluation in peripheral artery disease (PAD) by common vascular diagnostic tools is limited by open wounds, medial calcinosis or an altered collateral supply of the foot. Indocyanine green fluorescent imaging (ICG-FI) has recently been introduced as an alternative tool, but so far a standardized quantitative assessment of tissue perfusion in vascular surgery has not been performed for this purpose. The aim of this feasibility study was to investigate a new software for quantitative assessment of tissue perfusion in patients with PAD using indocyanine green fluorescent imaging (ICG-FI) before and after peripheral bypass grafting. Patients and methods: Indocyanine green fluorescent imaging was performed in seven patients using the SPY Elite system before and after peripheral bypass grafting for PAD (Rutherford III-VI). Visual and quantitative evaluation of tissue perfusion was assessed in an area of low perfusion (ALP) and high perfusion (AHP), each by three independent investigators. Data assessment was performed offline using a specially customized software package (Institute for Laser Technology, University Ulm, GmbH). Slope of fluorescent intensity (SFI) was measured as time-intensity curves. Values were compared to ankle-brachial index (ABI), slope of oscillation (SOO), and time to peak (TTP) obtained from photoplethysmography (PPG). Results: All measurements before and after surgery were successfully performed, showing that ABI, TTP, and SOO increased significantly compared to preoperative values, all being statistically significant (P < 0.05), except for TTP (p = 0.061). Further, SFI increased significantly in both ALP and AHP (P < 0.05) and correlated considerably with ABI, TTP, and SOO (P < 0.05). Conclusions: In addition to ABI and slope of oscillation (SOO), the ICG-FI technique allows visual assessment in combination with quantitative assessment of tissue perfusion in patients with PAD. Ratios related to different perfusion patterns and SFI seem to be useful tools to reduce factors disturbing ICG-FI measurements.


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