High-Pressure Freezing and Low-Temperature Processing of Plant Tissue Samples for Electron Microscopy

2013 ◽  
pp. 147-157 ◽  
Author(s):  
Ichirou Karahara ◽  
Byung-Ho Kang
Keyword(s):  
Electron Microscopy ◽  
High Pressure ◽  
Low Temperature ◽  
Plant Tissue ◽  
High Pressure Freezing ◽  
Tissue Samples ◽  
Low Temperature Processing

Electron Microscopy immunocytochemistry following cryofixation by high-pressure freezing and Low-temperature embedding

1993 ◽  
Vol 51 ◽  
pp. 112-113
Author(s):  
Kent McDonald ◽  
Mary K. Morphew ◽  
J.R. McIntosh
Keyword(s):  
Electron Microscopy ◽  
Developmental Biology ◽  
High Pressure ◽  
Low Temperature ◽  
Colloidal Gold ◽  
Room Temperature ◽  
High Pressure Freezing ◽  
Lr White ◽  
Lowicryl K4m

A look at recent journals publishing cell and developmental biology papers shows that localization of antigens by immunoEM techniques is becoming increasingly popular. One can also see that a wide variety of immunoEM methods are being applied: pre-embed and post-embed labelling, cryofixation and room temperature fixation, cryosections and resin sections, LR White and Lowicryl resins, 5 - 10 nm gold and silver-enhanced 1 nm gold, and so on. This array of choices can be confusing to investigators unfamiliar with EM techniques but wanting to do EM immunolocalization of their antibodies. A very helpful overview of the problems can be found in the article by Kellenberger and Hayat and in the series of books on colloidal gold methods edited by Hayat. In this presentation, we will consider the different ways to fix cells for immunoEM and our experience with LR White and Lowicryl K4M resins.

High-pressure freezing of cells in suspension for low-voltage scanning electron microscopy (LVSEM)

1991 ◽  
Vol 49 ◽  
pp. 64-65
Author(s):  
Marek Malecki ◽  
James Pawley ◽  
Hans Ris
Keyword(s):  
Electron Microscopy ◽  
High Pressure ◽  
Low Voltage ◽  
Culture Media ◽  
Cell Structure ◽  
Freeze Substitution ◽  
Ice Crystal ◽  
High Pressure Freezing ◽  
Cell Culture Media ◽  
Voltage Scanning

The ultrastructure of cells suspended in physiological fluids or cell culture media can only be studied if the living processes are stopped while the cells remain in suspension. Attachment of living cells to carrier surfaces to facilitate further processing for electron microscopy produces a rapid reorganization of cell structure eradicating most traces of the structures present when the cells were in suspension. The structure of cells in suspension can be immobilized by either chemical fixation or, much faster, by rapid freezing (cryo-immobilization). The fixation speed is particularly important in studies of cell surface reorganization over time. High pressure freezing provides conditions where specimens up to 500μm thick can be frozen in milliseconds without ice crystal damage. This volume is sufficient for cells to remain in suspension until frozen. However, special procedures are needed to assure that the unattached cells are not lost during subsequent processing for LVSEM or HVEM using freeze-substitution or freeze drying. We recently developed such a procedure.

Low Temperature Processing of Nano-Sized Magnesia Ceramics Using Ultra High Pressure

2013 ◽  
Vol 50 (3) ◽  
pp. 226-230
Author(s):  
Jeongho Song ◽  
Junghye Eom ◽  
Yunyoung Noh ◽  
Young-Wook Kim ◽  
Ohsung Song
Keyword(s):  
High Pressure ◽  
Low Temperature ◽  
Ultra High Pressure ◽  
Low Temperature Processing

Morphology of the lateral fields of the infective juvenile of the entomopathogenic family Steinernematidae (Nematoda) using high pressure freezing (HPF)

Nematology ◽  
2019 ◽  
Vol 22 (1) ◽  
pp. 69-74
Author(s):  
Zdeněk Mráček ◽  
Jiří Nermut’ ◽  
Martina Tesařová ◽  
Vladimír Půža
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
High Pressure ◽  
Infective Juvenile ◽  
Field Pattern ◽  
Taxonomic Character ◽  
High Pressure Freezing ◽  
Lateral Field ◽  
Transmission Electron ◽  
Scanning Electron

Summary The lateral field pattern of infective juveniles of the nematode family Steinernematidae is an important taxonomic character. Scanning electron microscopy (SEM) shows the number of ridges and lines or incisures clearly, but does not provide other details. In the present study, ten species from six clades of Steinernematidae have been studied for their lateral field morphology using SEM and high pressure freezing (HPF) with transmission electron microscopy (TEM). Both methods indicated the same number of ridges and lines, although HPF/TEM resulted in a more detailed morphology with differences between the species. The tips of the ridges are either finely rounded or pointed and the lines are V-shaped or have a broadened bottom. These characters represent an additional pattern that may be characteristic for some species within the phylogenetic clades. Further studies of the lateral field morphology of other species is needed to ascertain whether each pattern is clade specific and phylogenetically valuable.

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