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Since its introduction in 2017, MINFLUX nanoscopy has shown that it can visualise fluorescent molecules with an exceptional localisation precision of a few nanometres. In this overview, we provide a brief insight into technical implementations, fluorescent marker developments and biological studies that have been conducted in connection with MINFLUX imaging and tracking. We also formulate ideas on how MINFLUX nanoscopy and derived technologies could influence bioimaging in the future.

Super-resolution fluorescence microscopy for investigating bacterial cell biology.

Super-resolution fluorescence microscopy technologies developed over the past two decades have pushed the resolution limit for fluorescently labeled molecules into the nanometer range. These technologies have the potential to study bacterial structures, for example, macromolecular assemblies such as secretion systems, with single-molecule resolution on a millisecond time scale. Here we review recent applications of super-resolution fluorescence microscopy with a focus on bacterial secretion systems.

MINFLUX imaging of a bacterial molecular machine at nanometer resolution.

The resolution achievable with the established super-resolution fluorescence nanoscopy methods, such as STORM or STED, is in general not sufficient to resolve protein complexes or even individual proteins. Recently, minimal photon flux (MINFLUX) nanoscopy has been introduced that combines the strengths of STED and STORM nanoscopy and can achieve a localization precision of less than 5 nm. We established a generally applicable workflow for MINFLUX imaging and applied it for the first time to a bacterial molecular machine, i.

Live imaging of Yersinia translocon formation and immune recognition in host cells.

Yersinia enterocolitica employs a type three secretion system (T3SS) to translocate immunosuppressive effector proteins into host cells. To this end, the T3SS assembles a translocon/pore complex composed of the translocator proteins YopB and YopD in host cell membranes serving as an entry port for the effectors. The translocon is formed in a Yersinia-containing pre-phagosomal compartment that is connected to the extracellular space.

The Ustilago maydis repetitive effector Rsp3 blocks the antifungal activity of mannose-binding maize proteins.

To cause disease in maize, the biotrophic fungus Ustilago maydis secretes a large arsenal of effector proteins. Here, we functionally characterize the repetitive effector Rsp3 (repetitive secreted protein 3), which shows length polymorphisms in field isolates and is highly expressed during biotrophic stages. Rsp3 is required for virulence and anthocyanin accumulation.

Fungal encephalitis following bone marrow transplantation: clinical findings and prognosis.

Background: Central nervous system fungal infections (FI) are important complications and a cause of mortality in patients who receive hematopoietic stem cell transplantation (HSCT).

Aims: To study the clinical aspects of fungal encephalitis (FE).

Settings And Design: The study was carried out at the HSCT Center of the Hospital de Clínicas, Federal University of Paraná, Curitiba, Brazil.

Chronic inflammatory demyelinating polyradiculoneuropathy in chronic graft-versus-host disease following allogeneic hematopoietic stem cell transplantation: case report.

The chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an unusual but important complication of hematopoietic stem cell transplantation (HSCT) rarely reported to date. We describe a 17-year-old woman with a diagnosis of acute myeloid leukemia due to Fanconi's anemia who was submitted to allogeneic HSCT and developed CIDP as part of graft-versus-host disease. Investigation showed high cerebrospinal fluid protein; electrophysiological studies revealed sensory-motor demyelinating polyradiculoneuropathy; muscle and nerve biopsy were compatible with CIDP.

A novel missense mutation in the caveolin-3 gene in rippling muscle disease.

Rippling muscle disease (RMD) is a benign myopathy with symptoms and signs of muscular hyperirritability. We report a 17-year-old patient who presented with muscular hypertrophy, local mounding on percussion, and a rippling phenomenon. Needle electromyography showed electrical silence during the rippling phenomenon.

[Emery-Dreifuss muscular dystrophy: case report].

The Emery-Dreifuss muscular dystrophy is a form of muscular dystrophy that frequently presents early contractures and cardiac conduction defects, caused by emerin deficiency in the inner nuclear membrane of the muscular fibers. A 19-years-old man it presented muscle weakness and hypotrophy in the proximal upper and lower limbs, dysphagia and early contractures in elbows and ankles, with familiar history compatible with X-linked inheritance form. The investigation showed increased serum creatinekinase levels electrocardiogram had a first degree atrioventricular block and right bundle branch block normal electromyography and nerve conduction study muscle biopsy disclosed myopathic characteristics and nuclear protein immunohystochemical analysis showed deficiency of emerin.

Scab incidence on pig skin resulting from hot particle exposures under varying conditions.

External irradiations of pig skin using radioactive sources of less than 600 microm in the largest dimension (hot particles) were carried out. The objective of the study was to determine a threshold for scab induction. Hot particles used included fissioned 235U and activated 170Tm, 17SYb, and 46Sc with maximum beta-particle energies of approximately 1.

Scab diameters on pig skin resulting from hot particle exposures under varying conditions.

The National Council on Radiation Protection and Measurements (NCRP) in NCRP Report Number 106 recommended a limit of 10(10) beta particles emitted from radioactive particles with sizes less than 1 mm (hot particles) to prevent acute deep ulceration. This recommendation was determined, in part, by regressing the diameter of the scabs induced by fissioned 235UC2 hot particles as a function of the logarithm of the number of beta particles emitted from the sources for one study. To validate this recommendation and the approach used by the NCRP, external irradiations of pig skin using radioactive sources of less than 600 microm in the largest dimension were carried out.

Effective half-lives for patients administered radiolabeled antibodies and calculated dose to the public in close proximity to patients.

Effective half-lives (T(e)) for radiolabeled antibodies can be much longer than that of traditional radiopharmaceuticals, potentially resulting in larger doses to members of the public. Clearance-rate data from patients treated with radio-labeled antibodies (RABs) were obtained from ten institutions. Calculations were made to determine if a single- or bi-exponential clearance-rate model was statistically justified; the results indicated that the former model was justified for more than 95% of the data.

Tritium radiobiology and relative biological effectiveness.

During the past decade, a large number of radiobiological studies have become available for tritium--many of them focusing on the relative biological effectiveness of tritium beta rays. These and previous studies indicate that tritium in body water produces the same spectrum of radiogenic effects (e.g.

Severity of organ injury as a predictor of acute mortality for disparate patterns of absorbed dose distribution.

Nonuniform distribution of absorbed dose is frequently encountered in the irradiated mammal; the degree of nonuniform distribution is generally more severe as the size of the animal increases and the energy or penetrating power of the radiation decreases. However, acute mortality under these conditions, e.g.

A system for simultaneous exposure of small animals to 60-Hz electric and magnetic fields.

Equipment designed for simultaneous exposure of rodents to 60-Hz electric and magnetic fields is described. Three identical systems were constructed, each capable of continuous exposure of 256 rats or 640 mice to a nominal electric field at less than 50 kV/m, and to horizontal and vertical magnetic fields at less than 1 mT. Design features, construction details, and results of various tests of the systems are described.

Cytogenetic analysis of mouse metaphase II oocytes following exposure to tritiated water.

Female mice were given different dosages (0, 3.0, 7.5, 15.

Effects of low level radiation upon the hematopoietic stem cell: implications for leukemogenesis.

These studies have addressed firstly the effect of single small doses of x-rays upon murine hematopoietic stem cells to obtain a better estimate of the Dq. It is small, of the order of 20 rad. Secondly, a dose fractionation schedule that does not kill or perturb the kinetics of hemopoietic cell proliferation was sought in order to investigate the leukemogenic potential of low level radiation upon an unperturbed hemopoietic system.

Regulation of diffusion-chamber granulopoiesis by colony-stimulating factor.

Studies were undertaken to examine the role of colony-stimulating factor (CSF) in controlling murine peritoneal diffusion-chamber (DC) granulopoiesis. Serum CSF was unchanged for 1-3 days after 700-750 rad total body irradiation. Variable increments were noted on the fourth and seventh days after irradiation.

The tritium toxicity program in the Medical Department at Brookhaven National Laboratory.

A summary of all the findings to date is given in Table 2. It appears from this information that it is possible to detect somatic, cytogenetic, and genetic effects resulting from exposures at 33 to 100 times the mpc's for HTO. Similar effects also result from exposure to external gamma rays at an equivalent dose.

Sister chromatid exchanges in bone marrow cells of mice maintained on tritiated water.

The ability of tritium to induce sister chromatid exchanges (SCEs) has been investigated in male mice of the Hale-Stoner-Brookhaven strain maintained on drinking water containing 3.0 microCi/ml tritiated water (HTO). At selected intervals after 28-261 days of consuming HTO, the frequency of SCEs and the kinetics of cellular proliferation were measured in bone marrow cells of animals maintained on HTO, and in age-matched control groups, by 5-bromo-2'-deoxyuridine labelling methods.

Residual toxicity in hematopoietic cells following a single dose of methylnitrosourea.

The residual injury to the proliferation capability of hemopoietic stem cells (CFU-S) which results from their exposure to leukemogenic agents was evaluated in mice given a single leukemogenic dose of methyl nitrosourea (MNU 50 mg/kg body weight, i.v.).

The distribution of tritium among the amino acids of proteins obtained from mice exposed to tritiated water.

The distribution of tritium among the amino acids of serum proteins in mice chronically exposed to tritiated water was determined by ion exchange chromatography of the protein hydrolysate. The specific activity of nonexchangeable tritium in these amino acids relative to the specific activity of tritium in the tissue water of mice ranged from 0.04 for phenylalanine and threonine to 1.