An improved simple method of specimen preparation for replicas or scanning electron microscopy*

1971 ◽  
Vol 94 (2) ◽  
pp. 185-187 ◽  
Author(s):  
W. B. Watters ◽  
R. C. Buck
Author(s):  
Toshihiko Takita ◽  
Tomonori Naguro ◽  
Toshio Kameie ◽  
Akihiro Iino ◽  
Kichizo Yamamoto

Recently with the increase in advanced age population, the osteoporosis becomes the object of public attention in the field of orthopedics. The surface topography of the bone by scanning electron microscopy (SEM) is one of the most useful means to study the bone metabolism, that is considered to make clear the mechanism of the osteoporosis. Until today many specimen preparation methods for SEM have been reported. They are roughly classified into two; the anorganic preparation and the simple preparation. The former is suitable for observing mineralization, but has the demerit that the real surface of the bone can not be observed and, moreover, the samples prepared by this method are extremely fragile especially in the case of osteoporosis. On the other hand, the latter has the merit that the real information of the bone surface can be obtained, though it is difficult to recognize the functional situation of the bone.


Author(s):  
M. Yamada ◽  
K. Ueda ◽  
K. Kuboki ◽  
H. Matsushima ◽  
S. Joens

Use of variable Pressure SEMs is spreading among electron microscopists The variable Pressure SEM does not necessarily require specimen Preparation such as fixation, dehydration, coating, etc which have been required for conventional scanning electron microscopy. The variable Pressure SEM allows operating Pressure of 1˜270 Pa in specimen chamber It does not allow microscopy of water-containing specimens under a saturated vapor Pressure of water. Therefore, it may cause shrink or deformation of water-containing soft specimens such as plant cells due to evaporation of water. A solution to this Problem is to lower the specimen temperature and maintain saturated vapor Pressures of water at low as shown in Fig. 1 On this technique, there is a Published report of experiment to have sufficient signal to noise ratio for scondary electron imaging at a relatively long working distance using an environmental SEM. We report here a new low temperature microscopy of soft Plant cells using a variable Pressure SEM (Hitachi S-225ON).


Author(s):  
T. Inoué ◽  
H. Koike

Low temperature scanning electron microscopy (LTSEM) is useful to avoid artifacts such as deformation and extraction, because specimens are not subjected to chemical fixation, dehydration and critical-point drying. Since Echlin et al developed a LTSEM, many techniques and instruments have been reported for observing frozen materials. However, intracellular structures such as mitochondria and endoplasmic reticulum have been unobservable by the method because of the low resolving power and inadequate specimen preparation methods. Recently, we developed a low temperature SEM that attained high resolutions. In this study, we introduce highly magnified images obtained by the newly developed LTSEM, especially intracellular structures which have been rapidly frozen without chemical fixation.[Specimen preparations] Mouse pancreas and brown adipose tissues (BAT) were used as materials. After the tissues were removed and cut into small pieces, the specimen was placed on a cryo-tip and rapidly frozen in liquid propane using a rapid freezing apparatus (Eiko Engineering Co. Ltd., Japan). After the tips were mounted on the specimen stage of a precooled cryo-holder, the surface of the specimen was manually fractured by a razor blade in liquid nitrogen. The cryo-holder was then inserted into the specimen chamber of the SEM (ISI DS-130), and specimens were observed at the accelerating voltages of 5-8 kV. At first the surface was slightly covered with frost, but intracellular structures were gradually revealed as the frost began to sublimate. Gold was then coated on the specimen surface while tilting the holder at 45-90°. The holder was connected to a liquid nitrogen reservoir by means of a copper braid to maintain low temperature.


2021 ◽  
Author(s):  
Rowan Mclachlan ◽  
Ashruti Patel ◽  
Andrea G Grottoli

Coral morphology is influenced by genetics, the environment, or the interaction of both, and thus is highly variable. This protocol outlines a non-destructive and relatively simple method for measuring Scleractinian coral sub-corallite skeletal structures (such as the septa length, theca thickness, and corallite diameter, etc.) using digital images produced as a result of digital microscopy or from scanning electron microscopy. This method uses X and Y coordinates of points placed onto photomicrographs to automatically calculate the length and/or diameter of a variety of sub-corallite skeletal structures in the Scleractinian coral Porites lobata. However, this protocol can be easily adapted for other coral species - the only difference may be the specific skeletal structures that are measured (for example, not all coral species have a pronounced columella or pali, or even circular corallites). This protocol is adapted from the methods described in Forsman et al. (2015) & Tisthammer et al. (2018). There are 4 steps to this protocol: 1) Removal of Organic Tissue from Coral Skeletons 2) Imaging of Coral Skeletons 3) Photomicrograph Image Analysis 4) Calculation of Corallite Microstructure Size This protocol was written by Dr. Rowan McLachlan and was reviewed by Ashruti Patel and Dr. Andréa Grottoli. Acknowledgments Leica DMS 1000 and Scanning Electron Microscopy photomicrographs used in this protocol were acquired at the Subsurface Energy Materials Characterization and Analysis Laboratory (SEMCAL), School of Earth Sciences at The Ohio State University, Ohio, USA. I would like to thank Dr. Julie Sheets, Dr. Sue Welch, and Dr. David Cole for training me on the use of these instruments.


1998 ◽  
Vol 4 (S2) ◽  
pp. 40-41
Author(s):  
N.R. Smith ◽  
R.A. Quinta

A partnership has developed between the Microscope and Graphic Imaging Center (MAGIC) at California State University, Hayward and Ohlone Community College. The purpose of the collaboration is to develop a program to allow community college students to gain experience in preparing and viewing samples using scanning electron microscopy technology. The learning module involves students from the Ohlone College Biology Majors Program and student mentors from CSUH. An additional component is the introduction of under-represented students into a Biology Fellowship Program in which they also participate in the SEM learning module. Participants for these programs are selected on the basis of their interest and how this experience will benefit them as expressed in a one-page written essay. Ten students are selected to participate in the programs.The objectives of the learning module are to: 1) learn specimen preparation techniques and develop skills in SEM technology; 2) gain hands-on experience and develop some laboratory skills necessary for effective use of a SEM in studying biological specimens; 3) share the experience gained with peers at their home institution.


2002 ◽  
Vol 8 (I1) ◽  
pp. 20-20

Topic: Characterization of Non-Conductive or Charging Materials by Microbeam AnalysisThe goal of this topical conference is to present the state of the art for materials characterization of non-conductive or charging materials using microbeam analysis. Examples of charging materials include polymeric materials, ceramic materials, and photoresist materials in the microelectronic industry. Also, the characterization of biological specimens will be covered because they are prone to problems related to charging. These materials are of great technological importance and their characterization is still a great challenge because they charge when analyzed with an electron beam. The techniques of microbeam analysis that will be considered are: X-ray Microanalysis in the Electron Microprobe, Low Voltage Scanning Electron Microscopy, Environmental Scanning Electron Microscopy, Analytical Electron Microscopy with Field Emission Transmission Electron Microscopy, and Focused Ion Beam Milling for specimen preparation. World experts will present papers on these topics. Papers from this topical conference will be published in a special issue of Microscopy & Microanalysis.


2012 ◽  
Vol 535-537 ◽  
pp. 280-283 ◽  
Author(s):  
Hao Ran An ◽  
Feng Shi Cai ◽  
Xue Wei Wang ◽  
Zhi Hao Yuan

Different morphology CuO nanostructures, including platelets, flower-like were simply synthesized at 350 °C controlled by droplet on hydrophobic substrate. This is a simple method which does not require any template, catalyst, or surfactant but can control the morphology of CuO from platelets to flowerlike. The morphologies are strongly dependent on the volume of droplet. Scanning electron microscopy (SEM), Optical microscope and X-ray diffraction (XRD) were used to observe the morphology, crystallinity, and chemical composition of the CuO structures. Growth mechanisms for shape selective CuO synthesis were proposed based on these results.


Sign in / Sign up

Export Citation Format

Share Document