Frontiers in cardiovascular medicine, Feb 12, 2024
Introduction: Investigating coronary microvascular perfusion responses after myocardial infarctio... more Introduction: Investigating coronary microvascular perfusion responses after myocardial infarction (MI) would aid in the development of flow preserving therapies. Laser speckle contrast imaging (LSCI) is a powerful tool used for real-time, non-contact, full-field imaging of blood flow in various tissues/organs. However, its use in the beating heart has been limited due to motion artifacts. Methods: In this paper, we report the novel use of LSCI, combined with custom speckle analysis software (SpAn), to visualise and quantitate changes in ventricular perfusion in adult and aged mice undergoing ischaemia-reperfusion (IR) injury. The therapeutic benefit of inhibiting the actions of the proinflammatory cytokine interleukin-36 (IL-36) was also investigated using an IL-36 receptor antagonist (IL-36Ra). Results: Imaging from uncovered and covered regions of the left ventricle demonstrated that whilst part of the LSCI flux signal was derived from beating motion, a significant contributor to the flux signal came from ventricular microcirculatory blood flow. We show that a biphasic flux profile corresponding to diastolic and systolic phases of the cardiac cycle can be detected without mathematically processing the total flux data to denoise motion artifacts. Furthermore, perfusion responses to ischaemia and postischaemia were strong, reproducible and could easily be detected without the need to subtract motionrelated flux signals. LSCI also identified significantly poorer ventricular perfusion in injured aged mice following IR injury which markedly improved with IL-36Ra. Discussion: We therefore propose that LSCI of the heart is possible despite motion artifacts and may facilitate future investigations into the role of the coronary microcirculation in cardiovascular diseases and development of novel therapies.
Platelets play a fundamental role in hemostasis and thrombosis. They are also involved in patholo... more Platelets play a fundamental role in hemostasis and thrombosis. They are also involved in pathologic conditions resulting from blocked blood vessels, including myocardial infarction and ischemic stroke. Platelet adhesion, activation, and aggregation at sites of vascular injury are regulated by a diverse repertoire of tyrosine kinase-linked and G proteincoupled receptors. Src family kinases (SFKs) play a central role in initiating and propagating signaling from several platelet surface receptors; however, the underlying mechanism of how SFK activity is regulated in platelets remains unclear. CD148 is the only receptor-like protein tyrosine phosphatase identified in platelets to date. In the present study, we show that mutant mice lacking CD148 exhibited a bleeding tendency and defective arterial thrombosis. Basal SFK activity was found to be markedly reduced in CD148-deficient platelets, resulting in a global hyporesponsiveness to agonists that signal through SFKs, including collagen and fibrinogen. G protein-coupled receptor responses to thrombin and other agonists were also marginally reduced. These results highlight CD148 as a global regulator of platelet activation and a novel antithrombotic drug target. (Blood. 2009;113: 4942-4954)
Introduction: Opening occluded coronary arteries in patients with myocardial infarction (MI) dama... more Introduction: Opening occluded coronary arteries in patients with myocardial infarction (MI) damages the delicate coronary microvessels through a process called myocardial ischaemia-reperfusion injury. Although mesenchymal stromal cells (MSCs) have the potential to limit this injury, clinical success remains limited. This may be due to (i) poor MSC homing to the heart (ii) infused MSCs, even if derived from the same site, being a heterogeneous population with varying therapeutic efficacy and (iii) conventional 2D culture of MSCs decreasing their homing and beneficial properties. This study investigated whether 3D culture of two distinctly different bone marrow (BM)-derived MSC sub-populations could improve their homing and coronary vasculoprotective efficacy. Methods: Intravital imaging of the anaesthetised mouse beating heart was used to investigate the trafficking and microvascular protective effects of two clonallyderived BM-derived MSC lines, namely CD317 neg MSCs-Y201 and CD317 pos MSCs-Y202, cultured using conventional monolayer and 3D hanging drop methods. Results: 3D culture consistently improved the adhesive behaviour of MSCs-Y201 to various substrates in vitro. However, it was their differential ability to reduce neutrophil events within the coronary capillaries and improve ventricular perfusion in vivo that was most remarkable. Moreover, dual therapy combined with heparin further improved the vasculoprotection afforded by 3D cultured MSCs-Y201 by also modifying platelet as well as neutrophil recruitment, which subsequently led to the greatest salvage of viable myocardium. Therapeutic benefit could mechanistically be explained by reductions in coronary endothelial oxidative stress and intercellular adhesion molecule-1 (ICAM-1)/vascular cell adhesion molecule-1 (VCAM-1) expression. However, since this was noted by both 2D and 3D cultured MSCs-Y201, therapeutic benefit is likely explained by the fact that 3D cultured MSCs-Y201 were the most potent sub-population at reducing serum levels of several pro-inflammatory cytokines.
The cardiac interstitium comprises a heterogeneous population of cells relevant to adult heart mu... more The cardiac interstitium comprises a heterogeneous population of cells relevant to adult heart muscle homeostasis. The detailed embryonic origin, quantitative and qualitative composition of cardiac interstitial cells remains unknown. By means of cell lineage tracing technologies, we have mapped the cellular components of the murine developing interstitium from the embryo to postnatal stages. Our results demonstrate that embryonic epicardial-derived cells (EPDC) pioneer the colonization of the interstitial space in the developing heart, displaying a typical transmural patterning across cardiac chamber walls. This study indicates that most EPDC remain at the cardiac interstitium in the form of fibroblastic(-like) cells, dynamically interacting with other non-epicardial-derived cells (e.g. bone marrow-derived cells). Using a variety of in vitro assays, we have characterized subpopulations of EPDC following criteria related to their molecular profile and mobilization properties (cell adhesion; proteolytic activity). From our data we conclude that: 1) EPDC are the first cells that populate the cardiac interstitium, where they home and remain along adulthood; 2) EPDC display a unique proteolytic program from early stages of cardiac development and 3) these cells might be instrumental to the homing of other cell types late in development/postnatal life. In summary, our work provides new information on the biology of cardiac interstitial cells (most specially cardiac fibroblasts), offering clues to understand paracrine and/or autocrine signals, as well as the ECM-related mechanisms (protein deposition and proteolysis), involved in clinically relevant patophysiological phenomena like ventricular remodeling after myocardial infarction.
<p>HPC-7 cells express (A) CD11a, but not (B) CD11b or (C) CD11c. Flow cytometry also revea... more <p>HPC-7 cells express (A) CD11a, but not (B) CD11b or (C) CD11c. Flow cytometry also revealed expression of (D) CD18 and (E) CD49d on the surface of HPC-7. (F) Whole bone marrow was depleted for lineage postive cells and labelled for Sca-1, c-kit and either IgG, CD18 or CD49d. The gating used is shown in panel F (FL2: Sca-1; FL3: c-Kit). (G) KSL cells express cell surface CD49d, while CD18 could not be detected. (H) CD18 and CD49d could be identified by flow cytometry following cell permeabilization.</p
Journal of Gastroenterology and Hepatology, Aug 1, 2000
Background and Aims : We have previously demonstrated that 60% ethanol (EtOH) increases macromole... more Background and Aims : We have previously demonstrated that 60% ethanol (EtOH) increases macromolecular leakage and induces focal lesion formation in areas of permanent flow stasis within gastric mucosal vessels. Nitric oxide (NO) may prevent lesion formation by inhibiting leakage. This study used fluorescent in vivo microscopy to investigate (i) whether L‐arginine (NO precursor) prevented EtOH induced injury and (ii) the mechanisms of protection. Methods : Experiments were carried out on anaesthetized rats (hypnorm/diazepam) receiving either intra‐arterial fluorescein isothiocyanate–bovine serum albumin (0.2 mL/100 g), a marker for quantitating leakage, or Acridine red (0.1 mL/100 g) which labels leukocytes. Animals then received 100, 300 or 500 mg/kg L‐arginine (i.a.) followed by 60% EtOH or distilled water, topically applied to the gastric mucosa (n = 6 for each group). Vessel diameter, macromolecular leakage of labelled albumin from post capillary venules (PCV) and capillaries and leukocyte activity were quantitated using image analysis. Results : L‐Arginine (100 mg/kg) did not increase vessel diameter or prevent EtOH‐induced lesion formation and leakage. Both 300 mg/kg and 500 mg/kg alone induced significant and sustained increases in PCV diameter after 15 (P &lt; 0.01) and 5 min (P &lt; 0.001), respectively. Lesion formation was only prevented by 300 mg/kg L‐arginine, whereas 500 mg/kg exacerbated haemorrhagic lesion formation over the entire exposed mucosa. Neither 300 mg/kg nor 500 mg/kg L‐arginine prevented leakage following EtOH. No leukocyte activity was observed following EtOH with or without L‐arginine pretreatment. Conclusion : L‐Arginine (300 mg/kg) prevented lesion formation. The mechanism of protection probably involved the increased blood flow in the dilated PCV and not the inhibition of leakage. The combined effects of EtOH and the possible high NO levels exacerbate gastric mucosal damage despite the increases observed in vessel diameter.
Expression of CD151 in the MDA-MB-231/CD151(+), MDA-MB-231/CD151(-), MDA-MB-231/CD151rec and MDA-... more Expression of CD151 in the MDA-MB-231/CD151(+), MDA-MB-231/CD151(-), MDA-MB-231/CD151rec and MDA-MB-231/CD151-QRD cells. Shown means of fluorescence intensity (MFI) for a representative experiment. The following mAbs (both IgG1) were used: 4C5G-negative control; 5C11-anti-CD151.
Medical & Biological Engineering & Computing, 2000
It has previously been shown that it is possible to differentiate between squamous and columnar e... more It has previously been shown that it is possible to differentiate between squamous and columnar epithelia in rat and resected human tissues using an impedance probe to make in vitro measurements. This probe can be passed down an endoscope allowing measurements to be made in patients. However, the probe emerges parallel to the oesophageal wall, with little room to manoeuvre. The conditions of control required to give reliable readings have been investigated. The importance of pressure applied and the angle of approach to the oesophagus was assessed. Pressures in the range 26.6Pa to 46.3kPa and angles in the range 15-90 degrees were considered. In in vitro studies it was observed that it was possible to obtain consistent readings with pressures greater than 2.9kPa and with angles greater than 15 degrees between the probe and the oesophagus. These conditions can be achieved in vivo, and readings obtained from twelve patients are shown (45 readings on normal squamous, 34 on Barrett's oesophagus and 22 on stomach). At low frequencies (9.6-153.2kHz), a Mann-Whitney test shows a significant difference (p<O.O01) when comparing the means from squamous and columnar, and also when readings from Barrett's and normal gastric epithelia are compared (p< 0.001).
Introduction Aspirin and a potent platelet P2Y12 inhibitor, such as prasugrel or ticagrelor, are ... more Introduction Aspirin and a potent platelet P2Y12 inhibitor, such as prasugrel or ticagrelor, are not always sufficient to prevent thrombus formation in patients with ST-elevation MI (STEMI), leading to “slow flow” or “no reflow” effects after stenting. GPIIb/IIIa inhibitors, such as eptifibatide, may help in this setting, but are not used routinely due to their bleeding risk. GPVI has critical roles in thrombosis and a minimal role in haemostasis. Here we tested whether depletion of GPVI has effects on thrombus formation after MI in an animal model and investigated the effects of a novel platelet GPVI inhibitor, glenzocimab (a Fab fragment of a monoclonal antibody), on platelet activation and thrombus formation when combined with aspirin and ticagrelor. Methods We used intravital microscopy in a murine model of ST-elevation myocardial infarction and ischaemia-reperfusion injury to investigate microvascular thrombosis. We investigated the antithrombotic effects of adding glenzocimab ...
The ability to image biological tissues is critical to our understanding of a range of systems an... more The ability to image biological tissues is critical to our understanding of a range of systems and processes. In the case of in situ living tissue, such imaging is hampered by the innate mechanical properties of the tissue. In many cases, this provides challenges in how to process large amounts of image data which may contain aberrations from movement. Generally, current tools require the provision of reference images and are unable to maintain temporal correlations within an image set. Here, we describe a tool-Tify-which can accurately predict a numerical quality score versus human scoring and can analyse image sets in a manner that allows the maintenance of temporal relationships. The tool uses regression-based techniques to link image statistics to image quality based on user provided scores from a sample of images. Scores calculated by the software correlate strongly with the scores provided by human users. We identified that, in most cases, the software requires users to score between 20-30 frames in order to be able to accurately calculate the remaining images. Importantly, our results suggest that the software can use coefficients generated from consolidated image sets to process images without the need for additional manual scoring. Finally, the tool is able to use a frame windowing technique to identify the highest quality frame from a moving window, thus retaining macro-chronological connections between frames. In summary, Tify is able to successfully predict the quality of images in an image set based on a small number of sample scores provided by end-users. This software has the potential to improve the effectiveness of biological imaging techniques where motion artefacts, even in the presence of stabilisation, pose a significant problem.
Frontiers in cardiovascular medicine, Feb 12, 2024
Introduction: Investigating coronary microvascular perfusion responses after myocardial infarctio... more Introduction: Investigating coronary microvascular perfusion responses after myocardial infarction (MI) would aid in the development of flow preserving therapies. Laser speckle contrast imaging (LSCI) is a powerful tool used for real-time, non-contact, full-field imaging of blood flow in various tissues/organs. However, its use in the beating heart has been limited due to motion artifacts. Methods: In this paper, we report the novel use of LSCI, combined with custom speckle analysis software (SpAn), to visualise and quantitate changes in ventricular perfusion in adult and aged mice undergoing ischaemia-reperfusion (IR) injury. The therapeutic benefit of inhibiting the actions of the proinflammatory cytokine interleukin-36 (IL-36) was also investigated using an IL-36 receptor antagonist (IL-36Ra). Results: Imaging from uncovered and covered regions of the left ventricle demonstrated that whilst part of the LSCI flux signal was derived from beating motion, a significant contributor to the flux signal came from ventricular microcirculatory blood flow. We show that a biphasic flux profile corresponding to diastolic and systolic phases of the cardiac cycle can be detected without mathematically processing the total flux data to denoise motion artifacts. Furthermore, perfusion responses to ischaemia and postischaemia were strong, reproducible and could easily be detected without the need to subtract motionrelated flux signals. LSCI also identified significantly poorer ventricular perfusion in injured aged mice following IR injury which markedly improved with IL-36Ra. Discussion: We therefore propose that LSCI of the heart is possible despite motion artifacts and may facilitate future investigations into the role of the coronary microcirculation in cardiovascular diseases and development of novel therapies.
Platelets play a fundamental role in hemostasis and thrombosis. They are also involved in patholo... more Platelets play a fundamental role in hemostasis and thrombosis. They are also involved in pathologic conditions resulting from blocked blood vessels, including myocardial infarction and ischemic stroke. Platelet adhesion, activation, and aggregation at sites of vascular injury are regulated by a diverse repertoire of tyrosine kinase-linked and G proteincoupled receptors. Src family kinases (SFKs) play a central role in initiating and propagating signaling from several platelet surface receptors; however, the underlying mechanism of how SFK activity is regulated in platelets remains unclear. CD148 is the only receptor-like protein tyrosine phosphatase identified in platelets to date. In the present study, we show that mutant mice lacking CD148 exhibited a bleeding tendency and defective arterial thrombosis. Basal SFK activity was found to be markedly reduced in CD148-deficient platelets, resulting in a global hyporesponsiveness to agonists that signal through SFKs, including collagen and fibrinogen. G protein-coupled receptor responses to thrombin and other agonists were also marginally reduced. These results highlight CD148 as a global regulator of platelet activation and a novel antithrombotic drug target. (Blood. 2009;113: 4942-4954)
Introduction: Opening occluded coronary arteries in patients with myocardial infarction (MI) dama... more Introduction: Opening occluded coronary arteries in patients with myocardial infarction (MI) damages the delicate coronary microvessels through a process called myocardial ischaemia-reperfusion injury. Although mesenchymal stromal cells (MSCs) have the potential to limit this injury, clinical success remains limited. This may be due to (i) poor MSC homing to the heart (ii) infused MSCs, even if derived from the same site, being a heterogeneous population with varying therapeutic efficacy and (iii) conventional 2D culture of MSCs decreasing their homing and beneficial properties. This study investigated whether 3D culture of two distinctly different bone marrow (BM)-derived MSC sub-populations could improve their homing and coronary vasculoprotective efficacy. Methods: Intravital imaging of the anaesthetised mouse beating heart was used to investigate the trafficking and microvascular protective effects of two clonallyderived BM-derived MSC lines, namely CD317 neg MSCs-Y201 and CD317 pos MSCs-Y202, cultured using conventional monolayer and 3D hanging drop methods. Results: 3D culture consistently improved the adhesive behaviour of MSCs-Y201 to various substrates in vitro. However, it was their differential ability to reduce neutrophil events within the coronary capillaries and improve ventricular perfusion in vivo that was most remarkable. Moreover, dual therapy combined with heparin further improved the vasculoprotection afforded by 3D cultured MSCs-Y201 by also modifying platelet as well as neutrophil recruitment, which subsequently led to the greatest salvage of viable myocardium. Therapeutic benefit could mechanistically be explained by reductions in coronary endothelial oxidative stress and intercellular adhesion molecule-1 (ICAM-1)/vascular cell adhesion molecule-1 (VCAM-1) expression. However, since this was noted by both 2D and 3D cultured MSCs-Y201, therapeutic benefit is likely explained by the fact that 3D cultured MSCs-Y201 were the most potent sub-population at reducing serum levels of several pro-inflammatory cytokines.
The cardiac interstitium comprises a heterogeneous population of cells relevant to adult heart mu... more The cardiac interstitium comprises a heterogeneous population of cells relevant to adult heart muscle homeostasis. The detailed embryonic origin, quantitative and qualitative composition of cardiac interstitial cells remains unknown. By means of cell lineage tracing technologies, we have mapped the cellular components of the murine developing interstitium from the embryo to postnatal stages. Our results demonstrate that embryonic epicardial-derived cells (EPDC) pioneer the colonization of the interstitial space in the developing heart, displaying a typical transmural patterning across cardiac chamber walls. This study indicates that most EPDC remain at the cardiac interstitium in the form of fibroblastic(-like) cells, dynamically interacting with other non-epicardial-derived cells (e.g. bone marrow-derived cells). Using a variety of in vitro assays, we have characterized subpopulations of EPDC following criteria related to their molecular profile and mobilization properties (cell adhesion; proteolytic activity). From our data we conclude that: 1) EPDC are the first cells that populate the cardiac interstitium, where they home and remain along adulthood; 2) EPDC display a unique proteolytic program from early stages of cardiac development and 3) these cells might be instrumental to the homing of other cell types late in development/postnatal life. In summary, our work provides new information on the biology of cardiac interstitial cells (most specially cardiac fibroblasts), offering clues to understand paracrine and/or autocrine signals, as well as the ECM-related mechanisms (protein deposition and proteolysis), involved in clinically relevant patophysiological phenomena like ventricular remodeling after myocardial infarction.
<p>HPC-7 cells express (A) CD11a, but not (B) CD11b or (C) CD11c. Flow cytometry also revea... more <p>HPC-7 cells express (A) CD11a, but not (B) CD11b or (C) CD11c. Flow cytometry also revealed expression of (D) CD18 and (E) CD49d on the surface of HPC-7. (F) Whole bone marrow was depleted for lineage postive cells and labelled for Sca-1, c-kit and either IgG, CD18 or CD49d. The gating used is shown in panel F (FL2: Sca-1; FL3: c-Kit). (G) KSL cells express cell surface CD49d, while CD18 could not be detected. (H) CD18 and CD49d could be identified by flow cytometry following cell permeabilization.</p
Journal of Gastroenterology and Hepatology, Aug 1, 2000
Background and Aims : We have previously demonstrated that 60% ethanol (EtOH) increases macromole... more Background and Aims : We have previously demonstrated that 60% ethanol (EtOH) increases macromolecular leakage and induces focal lesion formation in areas of permanent flow stasis within gastric mucosal vessels. Nitric oxide (NO) may prevent lesion formation by inhibiting leakage. This study used fluorescent in vivo microscopy to investigate (i) whether L‐arginine (NO precursor) prevented EtOH induced injury and (ii) the mechanisms of protection. Methods : Experiments were carried out on anaesthetized rats (hypnorm/diazepam) receiving either intra‐arterial fluorescein isothiocyanate–bovine serum albumin (0.2 mL/100 g), a marker for quantitating leakage, or Acridine red (0.1 mL/100 g) which labels leukocytes. Animals then received 100, 300 or 500 mg/kg L‐arginine (i.a.) followed by 60% EtOH or distilled water, topically applied to the gastric mucosa (n = 6 for each group). Vessel diameter, macromolecular leakage of labelled albumin from post capillary venules (PCV) and capillaries and leukocyte activity were quantitated using image analysis. Results : L‐Arginine (100 mg/kg) did not increase vessel diameter or prevent EtOH‐induced lesion formation and leakage. Both 300 mg/kg and 500 mg/kg alone induced significant and sustained increases in PCV diameter after 15 (P &lt; 0.01) and 5 min (P &lt; 0.001), respectively. Lesion formation was only prevented by 300 mg/kg L‐arginine, whereas 500 mg/kg exacerbated haemorrhagic lesion formation over the entire exposed mucosa. Neither 300 mg/kg nor 500 mg/kg L‐arginine prevented leakage following EtOH. No leukocyte activity was observed following EtOH with or without L‐arginine pretreatment. Conclusion : L‐Arginine (300 mg/kg) prevented lesion formation. The mechanism of protection probably involved the increased blood flow in the dilated PCV and not the inhibition of leakage. The combined effects of EtOH and the possible high NO levels exacerbate gastric mucosal damage despite the increases observed in vessel diameter.
Expression of CD151 in the MDA-MB-231/CD151(+), MDA-MB-231/CD151(-), MDA-MB-231/CD151rec and MDA-... more Expression of CD151 in the MDA-MB-231/CD151(+), MDA-MB-231/CD151(-), MDA-MB-231/CD151rec and MDA-MB-231/CD151-QRD cells. Shown means of fluorescence intensity (MFI) for a representative experiment. The following mAbs (both IgG1) were used: 4C5G-negative control; 5C11-anti-CD151.
Medical & Biological Engineering & Computing, 2000
It has previously been shown that it is possible to differentiate between squamous and columnar e... more It has previously been shown that it is possible to differentiate between squamous and columnar epithelia in rat and resected human tissues using an impedance probe to make in vitro measurements. This probe can be passed down an endoscope allowing measurements to be made in patients. However, the probe emerges parallel to the oesophageal wall, with little room to manoeuvre. The conditions of control required to give reliable readings have been investigated. The importance of pressure applied and the angle of approach to the oesophagus was assessed. Pressures in the range 26.6Pa to 46.3kPa and angles in the range 15-90 degrees were considered. In in vitro studies it was observed that it was possible to obtain consistent readings with pressures greater than 2.9kPa and with angles greater than 15 degrees between the probe and the oesophagus. These conditions can be achieved in vivo, and readings obtained from twelve patients are shown (45 readings on normal squamous, 34 on Barrett's oesophagus and 22 on stomach). At low frequencies (9.6-153.2kHz), a Mann-Whitney test shows a significant difference (p<O.O01) when comparing the means from squamous and columnar, and also when readings from Barrett's and normal gastric epithelia are compared (p< 0.001).
Introduction Aspirin and a potent platelet P2Y12 inhibitor, such as prasugrel or ticagrelor, are ... more Introduction Aspirin and a potent platelet P2Y12 inhibitor, such as prasugrel or ticagrelor, are not always sufficient to prevent thrombus formation in patients with ST-elevation MI (STEMI), leading to “slow flow” or “no reflow” effects after stenting. GPIIb/IIIa inhibitors, such as eptifibatide, may help in this setting, but are not used routinely due to their bleeding risk. GPVI has critical roles in thrombosis and a minimal role in haemostasis. Here we tested whether depletion of GPVI has effects on thrombus formation after MI in an animal model and investigated the effects of a novel platelet GPVI inhibitor, glenzocimab (a Fab fragment of a monoclonal antibody), on platelet activation and thrombus formation when combined with aspirin and ticagrelor. Methods We used intravital microscopy in a murine model of ST-elevation myocardial infarction and ischaemia-reperfusion injury to investigate microvascular thrombosis. We investigated the antithrombotic effects of adding glenzocimab ...
The ability to image biological tissues is critical to our understanding of a range of systems an... more The ability to image biological tissues is critical to our understanding of a range of systems and processes. In the case of in situ living tissue, such imaging is hampered by the innate mechanical properties of the tissue. In many cases, this provides challenges in how to process large amounts of image data which may contain aberrations from movement. Generally, current tools require the provision of reference images and are unable to maintain temporal correlations within an image set. Here, we describe a tool-Tify-which can accurately predict a numerical quality score versus human scoring and can analyse image sets in a manner that allows the maintenance of temporal relationships. The tool uses regression-based techniques to link image statistics to image quality based on user provided scores from a sample of images. Scores calculated by the software correlate strongly with the scores provided by human users. We identified that, in most cases, the software requires users to score between 20-30 frames in order to be able to accurately calculate the remaining images. Importantly, our results suggest that the software can use coefficients generated from consolidated image sets to process images without the need for additional manual scoring. Finally, the tool is able to use a frame windowing technique to identify the highest quality frame from a moving window, thus retaining macro-chronological connections between frames. In summary, Tify is able to successfully predict the quality of images in an image set based on a small number of sample scores provided by end-users. This software has the potential to improve the effectiveness of biological imaging techniques where motion artefacts, even in the presence of stabilisation, pose a significant problem.
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