A flow cell imaging system combined diffraction and bright field microscopy

Author(s):  
Siqi Yuan ◽  
Yu Sa ◽  
Pengpeng Sun ◽  
Yuangguang Han ◽  
Jingwen Feng
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.


2021 ◽  
Vol 9 (8) ◽  
pp. 848
Author(s):  
Elise E. B. LaDouceur ◽  
Linda A. Kuhnz ◽  
Christina Biggs ◽  
Alicia Bitondo ◽  
Megan Olhasso ◽  
...  

Sea pigs (Scotoplanes spp.) are deep-sea dwelling sea cucumbers of the phylum Echinodermata, class Holothuroidea, and order Elasipodida. Few reports are available on the microscopic anatomy of these deep-sea animals. This study describes the histologic findings of two, wild, male and female Scotoplanes sp. collected from Monterey Bay, California. Microscopic findings were similar to other holothuroids, with a few notable exceptions. Sea pigs were bilaterally symmetrical with six pairs of greatly enlarged tube feet arising from the lateral body wall and oriented ventrally for walking. Neither a rete mirabile nor respiratory tree was identified, and the large tube feet may function in respiration. Dorsal papillae protrude from the bivium and are histologically similar to tube feet with a large, muscular water vascular canal in the center. There were 10 buccal tentacles, the epidermis of which was highly folded. Only a single gonad was present in each animal; both male and female had histologic evidence of active gametogenesis. In the male, a presumed protozoal cyst was identified in the aboral intestinal mucosa, and was histologically similar to previous reports of coccidians. This work provides control histology for future investigations of sea pigs and related animals using bright field microscopy.


2017 ◽  
pp. 307-344
Author(s):  
Onkar D. Dhingra ◽  
James B. Sinclair

2021 ◽  
Vol 44 (4) ◽  
Author(s):  
Fabio Giavazzi ◽  
Antara Pal ◽  
Roberto Cerbino

Abstract Soft and biological materials are often composed of elementary constituents exhibiting an incessant roto-translational motion at the microscopic scale. Tracking this motion with a bright-field microscope becomes increasingly challenging when the particle size becomes smaller than the microscope resolution, a case which is frequently encountered. Here we demonstrate squared-gradient differential dynamic microscopy (SG-DDM) as a tool to successfully use bright-field microscopy to extract the roto-translational dynamics of small anisotropic colloidal particles, whose rotational motion cannot be tracked accurately in direct space. We provide analytical justification and experimental demonstration of the method by successful application to an aqueous suspension of peanut-shaped particles. Graphic abstract


2004 ◽  
Vol 56 (3) ◽  
pp. 325-332 ◽  
Author(s):  
L.A. Barros ◽  
R. Tortelly ◽  
R.M. Pinto ◽  
D.C. Gomes

Rabbits were infected per os with 10 Eustrongylides ignotus L4 and with 50 Contracaecum multipapillatum L3 per rabbit, recovered from naturally infected freshwater fishes (Hoplias malabaricus) in order to evaluate the patogenicity of these two nematode species in mammalian host. Two rabbits (20%) infected with E. ignotus died before the fourth day post-inoculation (one after 51 and the other after 78 hours). Six rabbits (60%) were inappetent until the fifth day following experimental inoculation. No clinical signs in rabbits inoculated with C. multipapillatum were observed; nevertheless, eight (80%) animals were positive for this nematode species. Rabbits inoculated with E. ignotus, had gastric congestion with hematoma of the gastric wall in 60% of the cases. Peritoneum was congested in 20% of the animals with the presence of peritoneal abscess in 10% of the cases. All inoculated animals showed hyperemia of the gastric mucosa with hemorrhagic gastritis due to infections with E. ignotus. In C. multipapillatum inoculated animals, the hyperemia was followed by disruption of the epithelial mucosa in the sites of parasite attachment. In the gastric mucosa, miscellaneous leukocitary infiltrates, with multifocal necrosis reaching the submucosa in the infections with C. multipapillatum were observed under bright field microscopy. Perforating lesions in several organs, mainly in the gastric wall, pancreas and liver, always in the presence of a mixed inflammatory process, intensely fibrous, with hemorrhage and necrosis were observed in animals infected with E. ignotus.


2019 ◽  
Vol 16 (1) ◽  
Author(s):  
Yuki Takamatsu ◽  
Olga Dolnik ◽  
Takeshi Noda ◽  
Stephan Becker

Abstract Background Live-cell imaging is a powerful tool for visualization of the spatio-temporal dynamics of moving signals in living cells. Although this technique can be utilized to visualize nucleocapsid transport in Marburg virus (MARV)- or Ebola virus-infected cells, the experiments require biosafety level-4 (BSL-4) laboratories, which are restricted to trained and authorized individuals. Methods To overcome this limitation, we developed a live-cell imaging system to visualize MARV nucleocapsid-like structures using fluorescence-conjugated viral proteins, which can be conducted outside BSL-4 laboratories. Results Our experiments revealed that nucleocapsid-like structures have similar transport characteristics to those of nucleocapsids observed in MARV-infected cells, both of which are mediated by actin polymerization. Conclusions We developed a non-infectious live cell imaging system to visualize intracellular transport of MARV nucleocapsid-like structures. This system provides a safe platform to evaluate antiviral drugs that inhibit MARV nucleocapsid transport.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
C. Martins ◽  
A. P. Rodrigo ◽  
L. Cabrita ◽  
P. Henriques ◽  
A. J. Parola ◽  
...  

Abstract True green pigments in the animal kingdom are scarce and are almost invariably porphyrinoids. Endogenous porphyrins resulting from the breakdown of haem are usually known as “bile pigments”. The pigmentation of intertidal Polychaeta has long gained attention due to its variety and vivid patterning that often seems incompatible with camouflage, as it occurs with Eulalia viridis, one of the few truly green Polychaeta. The present study combined UV and bright-field microscopy with HPLC to address the presence and distribution of pigments in several organs. The results showed two major types of porphyrin-like pigments, yellowish and greenish in colour, that are chiefly stored as intraplasmatic granules. Whereas the proboscis holds yellow pigments, the skin harbours both types in highly specialised cells. In their turn, oocytes and intestine have mostly green pigments. Despite some inter-individual variation, the pigments tend to be stable after prolonged storage at −20 °C, which has important implications for future studies. The results show that, in a foraging predator of the intertidal where melanins are circumscribed to lining the nervous system, porphyrinoid pigments have a key role in protection against UV light, in sensing and even as chemical defence against foulants and predators, which represents a remarkable adaptive feature.


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