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

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 3D Imaging of mammalian cells with ion-abrasion scanning electron microscopy

2009 ◽  
Vol 166 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Jurgen A.W. Heymann ◽  
Dan Shi ◽  
Sang Kim ◽  
Donald Bliss ◽  
Jacqueline L.S. Milne ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Mammalian Cells ◽  
3D Imaging ◽  
Scanning Electron

scholarly journals Correlative and integrated light and electron microscopy of in-resin GFP fluorescence, used to localise diacylglycerol in mammalian cells

2014 ◽  
Vol 143 ◽  
pp. 3-14 ◽  
Author(s):  
Christopher J. Peddie ◽  
Ken Blight ◽  
Emma Wilson ◽  
Charlotte Melia ◽  
Jo Marrison ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Mammalian Cells ◽  
Light And Electron Microscopy ◽  
Gfp Fluorescence

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.

The Morphology of Vacuolated Cells in the Liver Following Administration of Vitamin A.

1973 ◽  
Vol 31 ◽  
pp. 408-409
Author(s):  
Odell T. Minick ◽  
Hidejiro Yokoo ◽  
Fawzia Batti
Keyword(s):  
Electron Microscopy ◽  
Body Weight ◽  
Vitamin A ◽  
Light And Electron Microscopy ◽  
Liver Cells ◽  
Prominent Feature ◽  
Vascular Space ◽  
Vacuolated Cell ◽  
Vacuolated Cells ◽  
Vacuolated Cytoplasm

Vacuolated cells in the liver of young rats were studied by light and electron microscopy following the administration of vitamin A (200 units per gram of body weight). Their characteristics were compared with similar cells found in untreated animals.In rats given vitamin A, cells with vacuolated cytoplasm were a prominent feature. These cells were found mostly in a perisinusoidal location, although some appeared to be in between liver cells (Fig. 1). Electron microscopy confirmed their location in Disse's space adjacent to the sinusoid and in recesses between liver cells. Some appeared to be bordering the lumen of the sinusoid, but careful observation usually revealed a tenuous endothelial process separating the vacuolated cell from the vascular space. In appropriate sections, fenestrations in the thin endothelial processes were noted (Fig. 2, arrow).

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