A Novel Microwave Device Designed to Preserve Cell Structure in Milliseconds

1990 ◽  
Vol 189 ◽  
Author(s):  
Gary R. Login ◽  
Susan Kissell ◽  
Barbara K. Dwyer ◽  
Ann M. Dvorak
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Cell Structure ◽  
Light And Electron Microscopy ◽  
Cell Suspensions ◽  
Microwave Device ◽  
Tissue Fixation ◽  
Microwave Exposure ◽  
Excellent Preservation ◽  
Reflected Power

ABSTRACTWe describe an innovative microwave instrument, designed in collaboration with and owned by Raytheon Company. The instrument permits the manipulation of biological specimens in their fluid milieu during the actual period of rapid tissue fixation. The specimen chamber is designed for sample containers up to 1.7 cm in diameter and 4.5 cm in height. Reflected power is reproducibly low, limiting the need for pretuning the microwave output to the sample. Microwave exposure can be controlled in 1 msecond increments with a range of 10 mseconds to 10 seconds. Mammalian cells and tissues fixed by this microwave device were evaluated by light and electron microscopy. Preliminary findings show large regions of excellent preservation in tissues and in cell suspensions in -100 mseconds.

scholarly journals Correlative 3D imaging of whole mammalian cells with light and electron microscopy

2011 ◽  
Vol 176 (3) ◽  
pp. 268-278 ◽  
Author(s):  
Gavin E. Murphy ◽  
Kedar Narayan ◽  
Bradley C. Lowekamp ◽  
Lisa M. Hartnell ◽  
Jurgen A.W. Heymann ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
3D Imaging ◽  
Light And Electron Microscopy

scholarly journals Distinguishing signal from autofluorescence in cryogenic correlated light and electron microscopy of mammalian cells

2017 ◽  
Author(s):  
Stephen D. Carter ◽  
Shrawan K. Mageswaran ◽  
Zachary J. Farino ◽  
João I. Mamede ◽  
Catherine M. Oikonomou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Fluorescent Protein ◽  
Organic Dyes ◽  
Fluorescent Proteins ◽  
Secretory Granules ◽  
Light And Electron Microscopy ◽  
Low Contrast ◽  
Fluorescent Protein Tags ◽  
Protein Tags

AbstractCryogenic correlated light and electron microscopy (cryo-CLEM) is a valuable tool for studying biological processes in situ. In cryo-CLEM, a target protein of interest is tagged with a fluorophore and the location of the corresponding fluorescent signal is used to identify the structure in low-contrast but feature-rich cryo-EM images. To date, cryo-CLEM studies of mammalian cells have relied on very bright organic dyes or fluorescent protein tags concentrated in virus particles. Here we describe a method to expand the application of cryo-CLEM to cells harboring genetically-encoded fluorescent proteins. We discovered that a variety of mammalian cells exhibit strong punctate autofluorescence when imaged under cryogenic conditions (80K). Compared to fluorescent protein tags, these sources of autofluorescence exhibit a broader spectrum of fluorescence, which we exploited to develop a simple, robust approach to discriminate between the two. We validate this method in INS-1 E cells using a mitochondrial marker, and apply it to study the ultrastructural variability of secretory granules in a near-native state within intact INS-1E pancreatic cells by high-resolution 3D electron cryotomography.

scholarly journals Correlative cryo super-resolution light and electron microscopy on mammalian cells using fluorescent proteins

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Maarten W. Tuijtel ◽  
Abraham J. Koster ◽  
Stefan Jakobs ◽  
Frank G. A. Faas ◽  
Thomas H. Sharp
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Fluorescent Proteins ◽  
Light And Electron Microscopy ◽  
Super Resolution

scholarly journals Distinguishing signal from autofluorescence in cryogenic correlated light and electron microscopy of mammalian cells

2018 ◽  
Vol 201 (1) ◽  
pp. 15-25 ◽  
Author(s):  
Stephen D. Carter ◽  
Shrawan K. Mageswaran ◽  
Zachary J. Farino ◽  
João I. Mamede ◽  
Catherine M. Oikonomou ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Light And Electron Microscopy

Improved filtration techniques for the concentration and cytological preservation of microalgae for electron microscopy

1973 ◽  
Vol 51 (2) ◽  
pp. 371-377 ◽  
Author(s):  
T. Bisalputra ◽  
J. Y. Cheng ◽  
F. J. R. Taylor ◽  
N. J. Antia
Keyword(s):  
Electron Microscopy ◽  
Natural Waters ◽  
Light And Electron Microscopy ◽  
Subsequent Removal ◽  
Freeze Etching ◽  
Excellent Preservation ◽  
Frequent Sampling ◽  
Convenient Preparation ◽  
Filtration Technique ◽  
Algal Cultures

A gentle filtration technique is described for concentration and fixation of phytoplankters directly on Millipore membranes, followed by subsequent removal of the membrane and embedment of the cellular material in epoxy resin. The technique shows excellent preservation of cell morphology, especially of fragile algal cells (dinoflagellates, zoospores, etc.), with little loss of material, and affords permanently fixed samples for both light and electron microscopy. It is ideally suitable for handling small amounts of algal cultures and for frequent sampling of such cultures in time-sequence studies. Adaptations of the technique are described which enable (1) high proportion of flagella preservation, (2) successful concentration of algal cells from water samples of low population density, and (3) convenient preparation of samples for "freeze-etching" or histochemical studies. A further application to scanning electron microscopy is described, where the replacement of Millipore membranes by Nuclepore filters affords the differentiation of nannoplanktonic cells from non-living debris and detritus, thereby holding promise for ecological studies of natural waters.

scholarly journals The istant changes of the salivary glands to stress

2010 ◽  
Vol 9 (1) ◽  
pp. 31-35
Author(s):  
A. V. Gerasimov ◽  
S. V. Logvinov ◽  
V. P. Kostyuchenko
Keyword(s):  
Electron Microscopy ◽  
Salivary Glands ◽  
Guinea Pigs ◽  
Light And Electron Microscopy ◽  
Secretory Activity ◽  
Duct Cell ◽  
Cell Stress ◽  
Microwave Exposure ◽  
Submandibular Salivary Glands ◽  
Granular Ducts

Night lighting and microwave exposure have been influence on structures of stress realization. The endocrine, fotoperiodical and adaptive functions of rodent submandibular salivary glands belonging to hormone produced duct cells. To evaluate their morphofunctional state at guinea-pigs and rats using the methods of light and electron microscopy there have been analyzed striated and granular ducts. It has been revealed that instant and twenty-four-hour strengthening of duct cell stress induced secretory activity was similar. It is concluded that salivary glands take part in circadian expectations to stress.

Hypobaric helium atmospheres for light and electron microscopy of mammalian cells

1976 ◽  
Vol 106 (3) ◽  
pp. 303-310
Author(s):  
Norman C. Lyon ◽  
Stanley E. Kraus ◽  
Donald F. Parsons
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Light And Electron Microscopy

scholarly journals Correlative fluorescence microscopy, transmission electron microscopy and secondary ion mass spectrometry (CLEM-SIMS) for cellular imaging

2020 ◽  
Author(s):  
Felix Lange ◽  
Paola Agüi-Gonzalez ◽  
Dietmar Riedel ◽  
Nhu T.N. Phan ◽  
Stefan Jakobs ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mass Spectrometry ◽  
Fluorescence Microscopy ◽  
High Performance ◽  
Secondary Ion Mass Spectrometry ◽  
Cell Structure ◽  
Light And Electron Microscopy ◽  
Epifluorescence Microscopy ◽  
Ion Mass Spectrometry ◽  
Secondary Ion

AbstractElectron microscopy (EM) has been employed for decades to analyze cell structure. To also analyze the positions and functions of specific proteins, one typically relies on immuno-EM or on a correlation with fluorescence microscopy, in the form of correlated light and electron microscopy (CLEM). Nevertheless, neither of these procedures is able to also address the isotopic composition of cells. To solve this, a correlation with secondary ion mass spectrometry (SIMS) would be necessary. SIMS has been correlated in the past to EM or to fluorescence microscopy in biological samples, but not to CLEM. We achieved this here, using a protocol based on transmission EM, conventional epifluorescence microscopy and nanoSIMS. The protocol is easily applied, and enables the use of all three technologies at high performance parameters. We suggest that CLEM-SIMS will provide substantial information that is currently beyond the scope of conventional correlative approaches.

Sign in / Sign up

Export Citation Format

Share Document