ABSTRACT
Super-resolution optical fluctuation imaging provides a resolution beyond the diffraction limit by analysing stochastic fluorescence fluctuations with higher-order statistics. Using nth order spatio-temporal cross-cumulants the spatial resolution and the sampling can be increased up to n-fold in all spatial dimensions. In this study, we extend the cumulant analysis into the spectral domain and propose a multicolor super-resolution scheme. The simultaneous acquisition of two spectral channels followed by spectral cross-cumulant analysis and unmixing increases the spectral sampling. The number of discriminable fluorophore species is thus not limited to the number of physical detection channels. Using two color channels, we demonstrate spectral unmixing of three fluorophore species in simulations and experiments in fixed and live cells. Based on an eigenvalue/vector analysis, we propose a scheme for an optimized spectral filter choice. Overall, our methodology provides a route for easy-to-implement multicolor sub-diffraction imaging using standard microscopes while conserving the spatial super-resolution property.
ABSTRACT
Super-resolution microscopy opened diverse new avenues of research by overcoming the resolution limit imposed by diffraction. Exploitation of the fluorescent emission of individual fluorophores made it possible to reveal structures beyond the diffraction limit. To accurately determine the resolution achieved during imaging is challenging with existing metrics. Here, we propose a method for assessing the resolution of individual super-resolved images based on image partial phase autocorrelation. The algorithm is model-free and does not require any user-defined parameters. We demonstrate its performance on a wide variety of imaging modalities, including diffraction-limited techniques. Finally, we show how our method can be used to optimize image acquisition and post-processing in super-resolution microscopy.
Subject(s)
Algorithms , Cells/ultrastructure , Image Processing, Computer-Assisted/standards , Microscopy, Confocal/methods , Microscopy, Fluorescence/methods , Animals , COS Cells , Chlorocebus aethiops , Computer Simulation , Fluorescent Dyes/chemistry , HeLa Cells , Humans , Image Processing, Computer-Assisted/methodsABSTRACT
Even though the incidence of toxic epidermal necrolysis (TEN) is low, it is also associated with a high mortality rate. The condition predominantly affects the skin, but may also affect the gastrointestinal tract, dramatically increasing mortality. We present a case of perforated sigmoid diverticulitis in the presence of TEN. The patient was taking medication, known to be a risk factor, and presented an affected total body surface area and temporal development similar to previously reported cases of TEN. Characteristic abdominal symptoms, however, were missing. Gastrointestinal involvement in TEN appears to be a poor prognostic factor; medical staff must therefore be alert to patients with TEN who complain of abdominal discomfort. The exact pathogenesis, however, remains unclear.
Subject(s)
Hepatic Encephalopathy/diagnosis , Hernia, Umbilical/etiology , Liver Cirrhosis/diagnosis , Emergencies , Female , Hepatic Encephalopathy/complications , Hernia, Umbilical/diagnosis , Hernia, Umbilical/surgery , Humans , Liver Cirrhosis/complications , Middle Aged , Prognosis , Rupture, SpontaneousSubject(s)
Colonic Pseudo-Obstruction/diagnosis , Hypothyroidism/complications , Acute Disease , Aged, 80 and over , Cholinesterase Inhibitors/therapeutic use , Colonic Pseudo-Obstruction/complications , Colonic Pseudo-Obstruction/diagnostic imaging , Colonic Pseudo-Obstruction/drug therapy , Colonic Pseudo-Obstruction/surgery , Colonic Pseudo-Obstruction/therapy , Diagnosis, Differential , Humans , Laparotomy , Male , Neostigmine/therapeutic use , Parasympatholytics/therapeutic use , Radiography, Abdominal , Radiography, Thoracic , Treatment OutcomeSubject(s)
Dyspepsia/etiology , Gallstones/diagnosis , Ileal Diseases/diagnosis , Ileus/diagnosis , Vomiting/etiology , Abdominal Pain/etiology , Aged, 80 and over , Female , Gallstones/surgery , Humans , Ileal Diseases/surgery , Ileus/surgery , Postoperative Complications/diagnosis , Postoperative Complications/surgery , Recurrence , Reoperation , Tomography, X-Ray ComputedABSTRACT
We present a case of a 65 year old female patient with diverticulitis. We performed a Hartmann's operation. The left paraduodenal hernia was diagnosed during the procedure and we incised the hernia sack. It was only retrospectively, that we saw the hernia on the preoperative CT scan pictures.