Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination.

Significance: Widefield microscopy of the entire dorsal part of mouse cerebral cortex enables large-scale ("mesoscopic") imaging of different aspects of neuronal activity with spectrally compatible fluorescent indicators as well as hemodynamics via oxy- and deoxyhemoglobin absorption. Versatile and cost-effective imaging systems are needed for large-scale, color-multiplexed imaging of multiple fluorescent and intrinsic contrasts. Aim: We aim to develop a system for mesoscopic imaging of two fluorescent and two reflectance channels. Approach: Excitation of red and green fluorescence is achieved through epi-illumination. Hemoglobin absorption imaging is achieved using 525- and 625-nm light-emitting diodes positioned around the objective lens. An aluminum hemisphere placed between objective and cranial window provides diffuse illumination of the brain. Signals are recorded sequentially by a single sCMOS detector. Results: We demonstrate the performance of our imaging system by recording large-scale spontaneous and stimulus-evoked neuronal, cholinergic, and hemodynamic activity in awake, head-fixed mice with a curved "crystal skull" window expressing the red calcium indicator jRGECO1a and the green acetylcholine sensor GRAB ACh 3.0 . Shielding of illumination light through the aluminum hemisphere enables concurrent recording of pupil diameter changes. Conclusions: Our widefield microscope design with a single camera can be used to acquire multiple aspects of brain physiology and is compatible with behavioral readouts of pupil diameter.

Widefield in vivo imaging system with two fluorescence and two reflectance channels, a single sCMOS detector, and shielded illumination.

SIGNIFICANCE: Widefield microscopy of the entire dorsal part of mouse cerebral cortex enables large-scale (mesoscopic) imaging of neuronal activity with fluorescent indicators as well as hemodynamics via oxy- and deoxyhemoglobin absorption. Versatile and cost-effective imaging systems are needed for large-scale, color-multiplexed imaging of multiple fluorescent and intrinsic contrasts. AIM: Develop a system for mesoscopic imaging of two fluorescent and two reflectance channels. APPROACH: Excitation of red and green fluorescence is achieved through epi-illumination. Hemoglobin absorption imaging is achieved using 525- and 625nm LEDs positioned around the objective lens. An aluminum hemisphere placed between objective and cranial window provides diffuse illumination of the brain. Signals are recorded sequentially by a single sCMOS detector. RESULTS: We demonstrate performance of our imaging system by recording large-scale spontaneous and stimulus-evoked neuronal, cholinergic, and hemodynamic activity in awake head-fixed mice with a curved crystal skull window expressing the red calcium indicator jRGECO1a and the green acetylcholine sensor GRABACh3.0 . Shielding of illumination light through the aluminum hemisphere enables concurrent recording of pupil diameter changes. CONCLUSIONS: Our widefield microscope design with single camera can be used to acquire multiple aspects of brain physiology and is compatible with behavioral readouts of pupil diameter.

[Experimental model for cardiogenic shock with pericardial tamponade]. / A cardiogen sokk modellezése pericardialis tamponáddal.

INTRODUCTION: Pericardial tamponade (PT) is a life-threatening condition, with low cardiac output. The hemodynamic consequences of PT can severely affect the circulation of all tissues, including the microcirculation of the kidneys and the intestinal mucosa. Our aim was to develop a hemodynamically stable and controllable large animal model of PT to study the consequences of cardiogenic shock. METHODS: Two groups of anesthetized vietnamese minipigs (n = 6, both groups) were used. Following laparotomy, a cannula was fixed into the pericardium through the diaphragm without thoracotomy. A sham-operated group served as control, in the second group 60-min PT was induced by intrapericardial injection of heparinised own blood. Throughout PT and 180-min reperfusion, macrohemodynamics, renal circulation and mesenteric macro- and microcirculation were monitored. Myeloperoxidase (MPO) activity was measured and in vivo histology was performed by confocal laser scanning endomicroscopy. RESULTS: The PT increased central venous pressure, heart rate and decreased mean arterial pressure, mesenteric flow (from 355.5 ± 112.4 vs 182.0 ± 59.1 ml/min) and renal arterial flow (from 159.63 ± 50.7 vs 35.902 ± 27.9 ml/min) and the microcirculation of the ileum. Elevated MPO activity (3.66 ± 1.6 vs 7.01 ± 1.44 mU/mg protein) and injury of the ileal mucosa were present also. SUMMARY: The reproducible large animal model is suitable for clinically relevant investigations of the hemodynamic and biochemical consequences of PT.