RESUMO
In this work a handheld Fluorescent Lifetime IMaging (FLIM) system based on a distally mounted < 2 mm2 128 × 120 single photon avalanche diode (SPAD) array operating over a > 1 m long wired interface is demonstrated. The head of the system is â¼4.5â cm x 4.5â cm x 4.5â cm making it suitable for hand-held ex vivo applications. This is, to the best of the authors' knowledge, the first example of a SPAD array mounted on the distal end of a handheld FLIM system in this manner. All existing systems to date use a fibre to collect and relay fluorescent light to detectors at the proximal end of the system. This has clear potential biological and biomedical applications. To demonstrate this, the system is used to provide contrast between regions of differing tissue composition in ovine kidney samples, and between healthy and stressed or damaged plant leaves. Additionally, FLIM videos are provided showing that frame rates of > 1â Hz are achievable. It is thus an important step in realising an in vivo miniaturized chip-on-tip FLIM endoscopy system.
Assuntos
Imagem Óptica , Fótons , Animais , Ovinos , Microscopia de Fluorescência/métodos , CorantesRESUMO
Following pathogen recognition, plant cells produce a nitrosative burst resulting in a striking increase in nitric oxide (NO), altering the redox state of the cell, which subsequently helps orchestrate a plethora of immune responses. NO is a potent redox cue, efficiently relayed between proteins through its co-valent attachment to highly specific, powerfully reactive protein cysteine (Cys) thiols, resulting in formation of protein S-nitrosothiols (SNOs). This process, known as S-nitrosylation, can modulate the function of target proteins, enabling responsiveness to cellular redox changes. Key targets of S-nitrosylation control the production of reactive oxygen species (ROS), the transcription of immune-response genes, the triggering of the hypersensitive response (HR) and the establishment of systemic acquired resistance (SAR). Here, we bring together recent advances in the control of plant immunity by S-nitrosylation, furthering our appreciation of how changes in cellular redox status reprogramme plant immune function.