A new method for searching intracellular stereo structures in plant tissues

1990 ◽  
Vol 48 (3) ◽  
pp. 720-721
Author(s):  
Yaqin Yao ◽  
Xuanli Jiang ◽  
Dongwei Hu
Keyword(s):  
Room Temperature ◽  
Plant Tissues ◽  
Specimen Preparation ◽  
List Type ◽  
Type Number ◽  
Cell Organelles ◽  
Mesophyll Cells ◽  
Thin Sections ◽  
Wheat Leaves ◽  
Ethanol Series

The procedure for specimen preparation consist of the following. 1.fixing the plant tissues routinely;2.washing the blocks thoroughly in PBS buffer;3.immersing the blocks in a PEG (polyethylene glycol 4000) series (50, 70, 90, 100%) and embedding in pure PEG at 50°C, and cooling slowly to room temperature;4.cutting into thin sections 5-10 nm thick by LKB-V microtome;5.washing the sections a few times in water, dehydrating in a graded ethanol series, and drying in a critical-point dryer using liquid CO2;6.sputter-coating with gold palladium; and7.investigating in a Hitachi S-450 scanning electron microscope.By this method, the distinct, uninjured intracellular stereo structures can be conveniently found without Joss of cell organelles. Moreover, there are advantages to studying their stereo relationships between various components of cells. Using wheat leaves, one can find chloroplasts, mitochondria, nuclei, nucleoli, and parts of the cytoskeleton system in the mesophyll cells and their sizes and distributions directly.

scholarly journals Getting Epoxy Semi-Thin Sections to Stick to Glass Slides

2007 ◽  
Vol 15 (4) ◽  
pp. 52-52
Author(s):  
Gilbert (Gib) Ahlstrand
Keyword(s):  
Toluidine Blue ◽  
Room Temperature ◽  
Water Drop ◽  
List Type ◽  
Distilled Water ◽  
Type Number ◽  
Blow Down ◽  
Simple Method ◽  
Thin Sections ◽  
Glass Slides

Semi-thin sections don't always want to stick to glass slides, but subbing slides is usually not needed. For sections about 2 μm thick and no larger than 4 mm on a slide, this simple method works well for me:1)Clean 1x3 inch glass slides with an ethanol rinse, then air dry at room temperature or blow down with a hair dryer.2)Collect sections on a drop or two of distilled water on the slide, transferred there from the microtome with a clean fine tipped artists brush. Collect about 8-12 sections per drop.3)Warm the slide beneath the water drop from below using an alcohol lamp, fairly hot, but not to boil, of course. After drying by heating the sections stick quite well.4)Stain, usually with 0.2 μm filtered toluidine blue, again heating but gently this time, for about a minute, until stain “develops” the section.5)Rinse that stain off with distilled water from a squirt bottle, even directing the spray right onto the sections to get rid of any precipitate. Dry again gently with flame.The heating is the trick. There should not be any need for subbing or otherwise treating slides other than cleaning them.

Hydration Chamber for Jeolco 200 Kv Microscope

1970 ◽  
Vol 28 ◽  
pp. 542-543
Author(s):  
V. R. Matricardi ◽  
G. G. Hausner ◽  
D. F. Parsons
Keyword(s):  
Electron Microscope ◽  
Water Vapor ◽  
Vapor Pressure ◽  
Pressure Gradient ◽  
Room Temperature ◽  
List Type ◽  
Type Number ◽  
Equilibrium Vapor Pressure

In order to observe room temperature hydrated specimens in an electron microscope, the following conditions should be satisfied: The specimen should be surrounded by water vapor as close as possible to the equilibrium vapor pressure corresponding to the temperature of the specimen.The specimen grid should be inserted, focused and photo graphed in the shortest possible time in order to minimize dehydration.The full area of the specimen grid should be visible in order to minimize the number of changes of specimen required.There should be no pressure gradient across the grid so that specimens can be straddled across holes.Leakage of water vapor to the column should be minimized.

Mitochondria microcrystals deposition in some inherited diseases of lipid metabolism.

1978 ◽  
Vol 36 (2) ◽  
pp. 256-257
Author(s):  
S. Laoussadi ◽  
A. Kahan ◽  
G. Aubouy ◽  
F. Delbarre
Keyword(s):  
Synovial Tissue ◽  
Storage Disease ◽  
Metabolic Diseases ◽  
Uranyl Acetate ◽  
List Type ◽  
Type Ii ◽  
Type Number ◽  
Lipid Storage Disease ◽  
Thin Sections ◽  
Mean Size

Several patients with Fabry's, Gaucher's diseases and hyperlipoproteinemia type II and with arthropatic manifestations were observed.As no histological explanation for these symptoms was available,an ultrastructural study of synovial tissue was done to establish an anatomoclinical relation.Material and Methods :synovial membrane samples were obtained by needle biopsies of the knee from three patients with arthropatic manifestations of each disease.They were fixed in 5% glutaraldehyde, postfixed in 1% osmium tetraoxyde and embedded in Epon 812. Thin sections coloured by uranyl acetate and lead citrate were observed with an Elmiskop I Siemens electron microscope.Two important phenomena were observed in synovial tissue:Specific patterns of each lipid storage disease,which are now well known.In all the three metabolic diseases, hydroxyapatite-like crystals were found. They are characterized by their intramitochondrial localization, without any relation with cristae,an anarchic disposition and a mean size of 550 A.Crystals may be found also free in the cytoplasm of synoviocytes Some micrographs suggest an evolution in four steps :a. mitochondria with only a few microcrystalsb. mitochondria stuffed with these structuresc. disruption of mitochondria membranesd. microcrystals appear free in the cytoplasm

scholarly journals A New Universal Acrylic Embedding Resin for Both Light and Electron Microcopy

1994 ◽  
Vol 2 (4) ◽  
pp. 21-22
Author(s):  
Donald P. Cox
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Light Scattering ◽  
Tissue Structure ◽  
Original Structure ◽  
Plant Tissues ◽  
List Type ◽  
Type Number ◽  
Tissue Specimen ◽  
Low Viscosity

Successful immunolabeling in electron microscopy of animal and plant tissues requires a combination of excellent antigen preservation while maintaining the original structure of the tissue. One important element is tissue embedding which accomplishes two goals for the immunohistochemist, the preservation of tissue specimen structure and maintenance of biological antigenicity. Tissue embedding in plastic resins is a common method in which several important elements must be considered.1.Fine tissue structure must not be damaged by the polymerization.2.The plastic must be stable to the electron beam.3.Light scattering properties of the plastic should be minimal.4.The plastic should cut easily.5.The plastic must be of sufficiently low viscosity to infiltrate the tissue.

scholarly journals Processing Cochlea For Paraffin Sectioning And SEM

1998 ◽  
Vol 6 (5) ◽  
pp. 28-29
Author(s):  
Donna C. Montague
Keyword(s):  
Guinea Pig ◽  
Formic Acid ◽  
Room Temperature ◽  
Test Tube ◽  
List Type ◽  
Distilled Water ◽  
Type Number ◽  
Guinea Pig Cochlea ◽  
Specimen Volume ◽  
Gentle Agitation

We have successfully sectioned decalcified paraffin embedded “inner ears” from mice, rats and humans. We have also prepared guinea pig cochlea for SEM, So, briefly:Paraffin:1)Immediately after dissection, place the cochlea in 10% neutral buffered formalin (NSF) and let fix with gentle agitation 24 hours per 2 mm thickness at room temperature.2)Decalcify by immersing in 5% formic acid at least 4X specimen volume.3)Gently agitate at room temperature and change at least once per 24 hours. Check for completeness of decalcification by decanting 4 mLs of “spent” formic acid into a clean, clear test tube, Add 1 mL of 5% ammonium oxaiate in distilled water. Let stand undisturbed for 15 to 20 minutes, White precipitate of calcium oxaiate indicates that the sample is not completely decalcified, Change formic acid at least once per 24 hours until the spent formic tests clear.

Effects of Heat Shock on Nucleolar Organization in Tobacco Cells

1988 ◽  
Vol 46 ◽  
pp. 264-265
Author(s):  
Y. H. Yen ◽  
Y. R. Chen
Keyword(s):  
Heat Shock ◽  
Cultured Cells ◽  
Uranyl Acetate ◽  
Cell Organelles ◽  
Step Method ◽  
Thin Sections ◽  
Culture Cells ◽  
Nucleolar Organization ◽  
Root Meristematic Cells ◽  
Ethanol Series

In the previous study, supraoptimal and lethal temperatures affected on the structure of cell organelles in root meristematic cells of soybean were reported(l). In the present study, nucleolar organization in cultured cells responsing to heat shock will be invastigated.Suspension culture cells of tobacco(Nicotiana tobaccum W 38) growing in MS medium containing 1 ppm 2,4-D and 1 ppm kinetin were divided into several groupes and incubated as follow: 1), 28°C(2h); 2), 38°C(2h); 3), 42°C(2h); 4), 45°C(2h); 5), actino- mycin D(2 ug/ml, 3h). After incubation, cells were collected, fixed in 3% glutaraldehyde, post-fixed in 1% osmic tetraoxide, dehydrated in ethanol series and embedded in Spurr's resin. Thin sections were collected on formvar-supported grids and stained with uranyl acetate and lead citrate. Silver-staining adopted the method of Medina et al.(2) and sample were prefixed in 3% glutaraldehyde, post-fixed in Carnoy's solution, and then impregnated with silver by 2-step method.

Cryotransfer device for cryo-SEM to observe unfixed frozen biological samples

1988 ◽  
Vol 46 ◽  
pp. 106-107
Author(s):  
T. Makita ◽  
T. Asahina ◽  
K. Manba ◽  
S. Kagabu
Keyword(s):  
Specimen Preparation ◽  
Rapid Freezing ◽  
List Type ◽  
Type Number ◽  
Thin Specimen ◽  
Specimen Holder ◽  
Solid Nitrogen ◽  
Check Points ◽  
Semi Solid ◽  
Transfer Device

Further to our previous reports on the combination of CRYO-SEM and WET-SEM, this is application of another type of cryo-SEM device(SP2000 Sputter cryo, ST2000 Semtran, and SC2000 Semcool of Emscope Lab,England) attached to WET- SEM(W-40,Akashi Beam Technol.Tokyo) and SEM(DS130,ABT.Tokyo) to observe unfixed tissues of rats. The main check points of this system were how its cryotransfer device could prevent frost and ample space of cryo chamber which is independent to SEM chamber could safely utilized.Briefly preparation steps before observation were as follows;To insert thin specimen into two metalic tubies and install this assembly in specimen holderFor rapid freezing immerse this whole assembly into semi-solid nitrogen.Enclose this frozen specimen within a small chamber of transfer device and evacuate the chamber.Transfer to precooled (less than-170°C) specimen preparation chamber.By a stroke of handle,crack two metalic tubes in which frozen samples are contained. Alternatively scrach the surface of samples by a surgical knife.

Practical Aspects of Preparing Thin Sections From Catalyst Specimens

1985 ◽  
Vol 43 ◽  
pp. 174-175
Author(s):  
I. Y. Chan ◽  
J. H. Cain ◽  
T. M. Rea
Keyword(s):  
Preparation Method ◽  
Image Resolution ◽  
Catalyst Characterization ◽  
Specimen Preparation ◽  
List Type ◽  
Thin Sections ◽  
Transmission Electron ◽  
Thin Sectioning ◽  
Thickness Variations ◽  
Number Of Publications

Transmission electron microscopy is an important technique for catalyst characterization. The thin-sectioning technique has been recognized as the preferable specimen preparation method in most cases. The merit of this technique has been discussed in a number of publications. It has been applied to catalysts containing alumina, carbon, silica, and zeolites. In this paper, we present the practical aspects we have found for obtaining high quality thin sections consistently.Requirements1.The sections should be mechanically and electrically stable under the electron beam.2.They should be as thin as possible in order to achieve the highest image resolution permitted by the instrument.3.The results should be readily reproducible so that comparison between similar samples can be made easily.4.The number of artifacts (e.g., those arising from shatter marks, knife marks, and thickness variations) should be minimized.

Systematics of Silicide Formation by High Dose Miplantation of Transition Metals into Si

1986 ◽  
Vol 74 ◽  
Author(s):  
F. Namavar ◽  
F. H. Sanchez ◽  
J. I. Budnick ◽  
A. H. Fasihuddin ◽  
H. C. Hayden
Keyword(s):  
Room Temperature ◽  
Hexagonal Structure ◽  
High Dose ◽  
List Type ◽  
Type Number ◽  
Silicide Formation ◽  
Metal Thin Films ◽  
X Ray ◽  
Cubic Structure ◽  
Fe Implantation

AbstractWe have systematically studied the formation of transition-metal thin films by high dose (up to 1018 ions/cm2) implantation of Ti, V, Cr, Mn, Fe, Co, Ni and Nb at room temperature and 350°C into Si <100>.For implantation at 350°C, our results, as obtained by Rutherford backscattering, X-ray diffractometry and Read Camera measurements, indicate that one can categorize these metals into two groups: 1.a chromium group which includes V, Cr, Nb, Ti and Mn. Metals V, Cr and Nb form compounds (VSi2, CrSi2. NbSi2) with a hexagonal structure of the CrSi2 type whereas Ti and Mn both form compounds (Ti5Si3, Mn5Si3) with a hexagonal structure of the Mn5Si2 type.2.an iron group which includes Fe, Co and Ni. These metals form compounds (FeSi, CoSi, NiSi) with a cubic structure of the FeSi type.In this paper the experimental results for Cr and Fe implantation at room temperature and 350°C will be discussed.

Solar Corona Investigation by the Coronal Line Photometer at Lomnický Peak

1994 ◽  
Vol 144 ◽  
pp. 431-434
Author(s):  
M. Minarovjech ◽  
M. Rybanský
Keyword(s):  
Solar Corona ◽  
Time Resolution ◽  
Active Regions ◽  
High Time ◽  
High Time Resolution ◽  
List Type ◽  
Type Number ◽  
Coronal Line ◽  
Global Cycle

AbstractThis paper deals with a possibility to use the ground-based method of observation in order to solve basic problems connected with the solar corona research. Namely:1.heating of the solar corona2.course of the global cycle in the corona3.rotation of the solar corona and development of active regions.There is stressed a possibility of high-time resolution of the coronal line photometer at Lomnický Peak coronal station, and use of the latter to obtain crucial observations.

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