Dynamic fluorescence lifetime sensing with CMOS single-photon avalanche diode arrays and deep learning processors

Measuring fluorescence lifetimes of fast-moving cells or particles have broad applications in biomedical sciences. This paper presents a dynamic fluorescence lifetime sensing (DFLS) system based on the time-correlated single-photon counting (TCSPC) principle. It integrates a CMOS 192 × 128 single-photon avalanche diode (SPAD) array, offering an enormous photon-counting throughput without pile-up effects. We also proposed a quantized convolutional neural network (QCNN) algorithm and designed a field-programmable gate array embedded processor for fluorescence lifetime determinations. The processor uses a simple architecture, showing unparallel advantages in accuracy, analysis speed, and power consumption. It can resolve fluorescence lifetimes against disturbing noise. We evaluated the DFLS system using fluorescence dyes and fluorophore-tagged microspheres. The system can effectively measure fluorescence lifetimes within a single exposure period of the SPAD sensor, paving the way for portable time-resolved devices and shows potential in various applications.     

Bacopa monnieri and its constituents is hypotensive in anaesthetized rats and vasodilator in various artery types

Ethnopharmacological relevance

Bacopa monnieri (Brahmi) provides traditional cognitive treatments possibly reflecting improved cerebral hemodynamics. Little is known about the cardiovascular actions of Brahmi. We sought to assess its effects on blood pressure and on isolated arteries, thus providing insights to clinical applications.

Materials and methods

Intravenous Brahmi (20-60 mg/kg) was tested on arterial blood pressure and heart rate of anaesthetized rats. In vitro vasorelaxation was assessed in arteries, with and without blockers of nitric oxide synthase (L-NAME), cyclooxygenase (indomethacin), and mechanical de-endothelialisation. The effects of Brahmi on Ca²⁺ influx and release from stores were investigated.

Results

Intravenous Brahmi extract (20-60 mg/kg) decreased systolic and diastolic pressures without affecting heart rate. Brahmi evoked relaxation in isolated arteries in order of potency: basilar (IC50 = 102 ± 16 μg/ml) > mesenteric (171 ± 31) > aortae (213 ± 68) > renal (IC50 = 375 ± 51) > tail artery (494 ± 93) > femoral arteries (>1000 μg/ml). Two saponins, bacoside A3 and bacopaside II, had similar vasodilator actions (IC50 = 8.3 ± 1.7 and 19.5 ± 6.3 μM). In aortae, without endothelium or in L-NAME (10-4 M), Brahmi was less potent (IC50 = 213 ± 68 to 2170 ± 664 and 1192 ± 167 μg/ml, respectively); indomethacin (10-5 M) was ineffective. In tail artery, Brahmi inhibited K⁺-depolarization induced Ca²⁺ influx and Ca²⁺ release from the sarcoplasmic reticulum by phenylephrine (10-5 M) or caffeine (20 mM).

Conclusions

Brahmi reduces blood pressure partly via releasing nitric oxide from the endothelium, with additional actions on vascular smooth muscle Ca²⁺ homeostasis. Some Brahmi ingredients could be efficacious antihypertensives and the vasodilation could account for some medicinal actions.     

Bacopa monnieri Increases Cerebral Blood Flow in Rat Independent of Blood Pressure

Bacopa monnieri (L.) Wettst. (Brahmi in India and Thailand) is an ayurvedic dementia treatment, but its effect on cerebral blood flow (CBF) is still unknown. We sought to test its chronic and acute effects on CBF compared with Ginkgo biloba and donepezil. CBF was measured by laser Doppler from rat cerebral cortex after 8 weeks of daily oral dosing of these drugs. Systolic blood pressure was also measured using the tail cuff method or via arterial cannulation. In rats treated with B. monnieri (40 mg/kg), CBF was 25% increased [2927 ± 123 perfusion units, (PU)] compared with shams (2337 ± 217 PU, p < 0.05, nine rats). G. biloba (60 mg/kg) also increased CBF (by 29% to 3019 ± 208 PU, p < 0.05, nine rats). No clear effect was obtained with donepezil (1 mg/kg). Chronic administration of the preparations had no effect on blood pressure. In contrast, intravenous acute infusion of these herbals (20–60 mg/kg) had marked dose‐dependent hypotensive actions (diastolic ~31 mmHg lower with 40 mg/kg of either extract), which correspondingly reduced CBF by ~15%. Likewise, CBF fell slightly with acute intravenous sodium nitroprusside and rose with noradrenaline. Donepezil (1 mg/kg) was slightly hypotensive without affecting CBF. Increased CBF with B. monnieri may account for its reported procognitive effect, and its further exploration as an alternative nootropic drug is worthwhile. Copyright © 2012 John Wiley & Sons, Ltd.     

Histogram clustering for rapid time-domain fluorescence lifetime image analysis

We propose a histogram clustering (HC) method to accelerate fluorescence lifetime imaging (FLIM) analysis in pixel-wise and global fitting modes. The proposed method’s principle was demonstrated, and the combinations of HC with traditional FLIM analysis were explained. We assessed HC methods with both simulated and experimental datasets. The results reveal that HC not only increases analysis speed (up to 106 times) but also enhances lifetime estimation accuracy. Fast lifetime analysis strategies were suggested with execution times around or below 30 μs per histograms on MATLAB R2016a, 64-bit with the Intel Celeron CPU (2950M @ 2GHz).