A simple method of preparing a cell suspension for scanning electron microscopy

1975 ◽  
Vol 31 (10) ◽  
pp. 1244-1246 ◽  
Author(s):  
G. E. Jones ◽  
R. Gillett
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.


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.


1971 ◽  
Vol 17 (6) ◽  
pp. 822-823 ◽  
Author(s):  
David J. Bibel ◽  
John W. Lawson

A simple procedure, using Millipore filters, for the observation of L-form colonies of streptococci by scanning electron microscopy is described.


Author(s):  
E.C. Chew ◽  
D.J. Riches ◽  
P.P.L. Tam ◽  
G.S.W. Tsao ◽  
T.K. Lam ◽  
...  

The use of microwave irradiation for fixation of human and animal tissue has been proven satisfactory at light microscopic and electron microscopic levels. The present communication reports the study of the same method of fixation of cell cultures for scanning electron microscopy.Trophoblasts were isolated from the placentae of mouse conceptuses at 10.5 days of gestation. The placenta was dissected out from the decidua and placed in Ca and Mg-free PBS, minced and then forced through a gauge-21 syringe needle. The tissue fragments were digested with 0.25% trypsin in Ca and Mg-free PBS for 20 - 30 minutes at 4°C. The digested tissue was then washed with complete PB1 medium. A single-cell suspension was obtained by spinning down the larger fragments by centrifugation. A known volume of the single-cell suspension was added to the culture medium (DCMEM and 20% FCS). The culture medium was changed after 24 hours to remove any unattached cells.


2017 ◽  
Vol 11 (6) ◽  
pp. 925-931 ◽  
Author(s):  
Yuta Kurashina ◽  
◽  
Shogo Miyata ◽  
Jun Komotori

A cell culture module capable of cooling stimulus to collect cells efficiently on a metal culture substrate was developed. We evaluated the cell collection ratio and morphology of the collected cells. Following a cooling stimulus (0°C) for 20 min, the number of collected cells was increased by 50% compared to that collected after trypsin treatment without pipetting from the metal culture substrate. Following the cooling stimulus, cells were observed by fluorescence microscopy and scanning electron microscopy; the cell filopodia were shrunken compared to non-cooling-stimulated cells. Furthermore, the combination of collagenase and cooling stimulation resulted in the collection of a comparable number of cells as that obtained using only trypsin. Thus, cell proliferation was improved compared to that following trypsin treatment. Therefore, this method can be applied for culturing cells that are susceptible to trypsin damage.


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