Light microscopy of wood using sanded surface instead of slides

IAWA Journal ◽  
2021 ◽  
pp. 1-14
Author(s):  
Peter Kitin ◽  
John C. Hermanson ◽  
Hisashi Abe ◽  
Satoshi Nakaba ◽  
Ryo Funada

Abstract Xylem anatomy is fundamental in studies of the evolution of terrestrial plants, tree ecophysiology, forestry, and wood science. Traditional xylem anatomical studies by light microscopy utilize wood sections. However, the procedures are laborious, and high-quality histological sections have been particularly challenging to achieve from recalcitrant wood species and dry wood material. Modern microscopy offers opportunities for speeding up the xylem anatomical preparations. In this regard, the merits of using a sanded surface for wood anatomical research have been largely overlooked. Sanding of wood surfaces is practiced in dendrochronology and wood identification studies exclusively for the investigation of macro features, such as tree rings, wood porosity, or parenchyma patterns. We conducted microscopic level investigations of sanded surfaces of difficult-to-section high-density woods such as Dalbergia and Quercus species by reflected white light and epifluorescence microscopy. Reflected white light or combinations of reflected light and fluorescence could clearly show xylem micro-features in sanded wood surfaces. The resolution of cell types after sanding with 1000-grit was similar to the resolution obtained by transmitted light microscopy in histological slides. The advantages of sanded wood surfaces compared to traditional wood sections can be summarized as cost- and time-effective sample preparation, large sample area, intact cell walls and tissue structure, preservation of chemical content and extractives, and even focus of the field of view. A simple procedure of wood sanding instead of microscopic slides can be used for xylem microscopy and automatic image analysis of xylem structure.

2014 ◽  
Vol 21 (1) ◽  
pp. 239-248 ◽  
Author(s):  
Ambroise Marin ◽  
Emmanuel Denimal ◽  
Stéphane Guyot ◽  
Ludovic Journaux ◽  
Paul Molin

AbstractIn biology, cell counting is a primary measurement and it is usually performed manually using hemocytometers such as Malassez blades. This work is tedious and can be automated using image processing. An algorithm based on Fourier transform filtering and the Hough transform was developed for Malassez blade grid extraction. This facilitates cell segmentation and counting within the grid. For the present work, a set of 137 images with high variability was processed. Grids were accurately detected in 98% of these images.


Author(s):  
Fred Y. Shen ◽  
Margaret M. Harrington ◽  
Logan A. Walker ◽  
Hon Pong Jimmy Cheng ◽  
Edward S. Boyden ◽  
...  

AbstractMapping neuroanatomy is a foundational goal towards understanding brain function. Electron microscopy (EM) has been the gold standard for connectivity analysis because nanoscale resolution is necessary to unambiguously resolve chemical and electrical synapses. However, molecular information that specifies cell types is often lost in EM reconstructions. To address this, we devised a light microscopy approach for connectivity analysis of defined cell types called spectral connectomics. We combined multicolor genetic labeling (Brainbow) of neurons with a multi-round immunostaining Expansion Microscopy (miriEx) strategy to simultaneously interrogate morphology, molecular markers, and connectivity in the same brain section. We applied our multimodal profiling strategy to directly link inhibitory neuron cell types with their network morphologies. Furthermore, we showed that correlative Brainbow and endogenous synaptic machinery immunostaining can be used to define putative synaptic connections between spectrally unique neurons, as well as map putative inhibitory and excitatory inputs. We envision that spectral connectomics can be applied routinely in neurobiology labs to gain insights into normal and pathophysiological neuroanatomy across multiple animals and time points.


1992 ◽  
Vol 103 (3) ◽  
pp. 823-830 ◽  
Author(s):  
S.T. Furlong ◽  
K.S. Thibault ◽  
R.A. Rogers

Schistosomes do not make sterols or fatty acids de novo and thus require host lipids for survival. The acquisition of host lipids may also be an important factor in the schistosome's defense from host immunity; however, little is known about the regulation of this process. Here we have examined binding of radiolabeled and fluorescently labeled liposomes to schistosomula, and followed incorporation of fluorescent phospholipids into the worm by both morphological and biochemical methods. Saturable binding of radiolabeled phosphatidylcholine containing liposomes was observed and epifluorescence microscopy showed binding of C6-NBD-phosphatidylcholine (C6-NBD-PC), C12-NBD-phosphatidylcholine (C12-NBD-PC) and C6-NBD-phosphatidyl-ethanolamine (C6-NBD-PE) containing liposomes on the surface of the parasite. Following back-exchange with unlabeled liposomes, NBD-PC and NBD-PE were observed to be preferentially incorporated into specific cell types within the worm. Furthermore, cells which had accumulated the fluorescent lipid formed an interconnecting cellular network immediately below the tegument, identified as cytons. By contrast, fluorescein-PE was found only on the surface of the parasite and in the gut but not in the cytons. Biochemical analysis demonstrated that > 90% of the C6-NBD-PC and C12-NBD-PC remained as the intact molecule after a one hour incubation with the parasite, but that greater than 70% of the NBD-PE was converted to other lipids. These studies demonstrate that incorporation of phospholipid analogs into schistosomula can be followed morphologically and biochemically. As there was little localization of NBD-PE or NBD-PC in the gut, these analogs must be assimilated by crossing the tegument.(ABSTRACT TRUNCATED AT 250 WORDS)


Life ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 40
Author(s):  
Kathryn Bywaters ◽  
Carol R. Stoker ◽  
Nelio Batista Do Nascimento ◽  
Lawrence Lemke

With the discovery of the persistent jets of water being ejected to space from Enceladus, an understanding of the effect of the space environment on potential organisms and biosignatures in them is necessary for planning life detection missions. We experimentally determine the survivability of microbial cells in liquid medium when ejected into vacuum. Epifluorescence microscopy, using a lipid stain, and SEM imaging were used to interrogate the cellular integrity of E. coli after ejected through a pressurized nozzle into a vacuum chamber. The experimental samples showed a 94% decrease in visible intact E. coli cells but showed a fluorescence residue in the shape of the sublimated droplets that indicated the presence of lipids. The differences in the experimental conditions versus those expected on Enceladus should not change the analog value because the process a sample would undergo when ejected into space was representative. E. coli was selected for testing although other cell types could vary physiologically which would affect their response to a vacuum environment. More testing is needed to determine the dynamic range in concentration of cells expected to survive the plume environment. However, these results suggest that lipids may be directly detectable evidence of life in icy world plumes.


2020 ◽  
Vol 147 ◽  
pp. 01001 ◽  
Author(s):  
Indah Istiqomah ◽  
Sukardi ◽  
Murwantoko ◽  
Alim Isnansetyo

Vibriosis is a bacterial disease that has been reported in Indonesian marine fish culture since the 1990s. The disease was reported mostly in grouper and shrimp (monodon and vanname) farming, although the infections in snapper (Lates calcarifer) and abalone (Haliotis squamata) aquaculture were also occurred. Causative agents of vibriosis in Indonesia marine fish is involving 14 species of vibrio namely Vibrio harveyi, V. anguillarum, V. alginoluticus, V. parahaemolyticus, V. fluvialis, V. furnisii, V. methcnikovii, V. vulnificus, V. ordalii, V. cincinnatiensis, V. carchariae, V. azureus, V. mimicus and V. damsela. Control of vibriosis is conducted with water quality management, applications of vaccines, antibiotics, probiotics and immunostimulants. Most vaccines developed and commercially available in Indonesia are in the form of inactive-whole cell types. The vaccine product is effective enough to protect fish from vibriosis. Probiotics have been widely studied since the 2000s and have been shown to increase fish and shrimp growth and resistances against vibriosis. Immunostimulants began to be developed since 2010 based on the extracts of terrestrial plants, seaweeds and chitosan. It is therefore concluded that the continuous existence of pathogenic strains in aquaculture farm requires further development of the control methods, including periodic updating of the vaccines, probiotics and immunostimulant formulas for more potent efficacies.


Parasitology ◽  
2013 ◽  
Vol 141 (4) ◽  
pp. 531-541 ◽  
Author(s):  
K. J. SOTO-ARREDONDO ◽  
L. L. FLORES-VILLAVICENCIO ◽  
J. J. SERRANO-LUNA ◽  
M. SHIBAYAMA ◽  
M. SABANERO-LÓPEZ

SUMMARYFree-living amoebae belonging to the genusAcanthamoebaare the causative agents of infections such as amoebic keratitis (AK), granulomatous amoebic encephalitis (GAE) and cutaneous lesions. The mechanisms involved in the establishment of infection are unknown. However, it is accepted that the initial phase of pathogenesis involves adherence to the host tissue. In this work, we analysed surface molecules with an affinity for epithelial and neuronal cells from the trophozoites ofAcanthamoeba castellanii. We also investigated the cellular mechanisms that govern the process of trophozoite adhesion to the host cells. We first used confocal and epifluorescence microscopy to examine the distribution of theA. castellaniiactin cytoskeleton during interaction with the host cells. The use of drugs, as cytochalasin B (CB) and latrunculin B (LB), revealed the participation of cytoskeletal filaments in the adhesion process. In addition, to identify the proteins and glycoproteins on the surface ofA. castellanii, the trophozoites were labelled with biotin and biotinylated lectins. The results revealed bands of surface proteins, some of which were glycoproteins with mannose andN-acetylglucosamine residues. Interaction assays of biotinylated amoebae proteins with epithelial and neuronal cells showed that some surface proteins had affinity for both cell types. The results of this study provide insight into the biochemical and cellular mechanisms of theAcanthamoebainfection process.


2006 ◽  
Author(s):  
Ran Yi ◽  
Kengyeh Chu ◽  
Jerome Mertz
Keyword(s):  

2019 ◽  
Vol 12 (1) ◽  
pp. 40-47 ◽  
Author(s):  
Andrew C. Jones ◽  
Nicholas M. Kearns ◽  
Jia-Jung Ho ◽  
Jessica T. Flach ◽  
Martin T. Zanni

2016 ◽  
Vol 55 (15) ◽  
pp. 4261 ◽  
Author(s):  
M. Blattmann ◽  
S. Kretschmer ◽  
S. Thiele ◽  
C. Ataman ◽  
H. Zappe ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document