[9] Visualization of Exocytosis by Quick Freezing and Freeze-Fracture

Carrie J. Merkle; Douglas E. Chandler

doi:10.1016/0076-6879(93)21011-V

[9] Visualization of Exocytosis by Quick Freezing and Freeze-Fracture

Carrie J. Merkle
, Douglas E. Chandler

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

This chapter discusses the visualization of exocytosis by quick freezing and freeze-fracture. Exocytosis occurs extremely rapidly. Quick freezing in combination with freeze-fracture offers advantages for visualizing exocytosis in that it captures events occurring within milliseconds after stimulation and reveals panoramic views of membranes. Cells frozen ultrarapidly—that is, by removing heat so fast that ice crystals are undetectable by electron microscopy (EM), can yield superbly preserved ultrastructure without membrane fusion-mimicking artifacts resulting from slow, selective chemical fixatives and membrane-disrupting cryoprotectants. The aluminum planchet to which specimens are frozen is designed to fit directly onto the Balzers (Nashua, NH) 301 specimen table and is secured with a screw cap. Alternatively, tables fitted with a pair of flange clamps can be custom designed for many freeze-fracture units, including the Balzers 400 model.

Original language	English (US)
Pages (from-to)	112-123
Number of pages	12
Journal	Methods in Enzymology
Volume	221
Issue number	C
DOIs	https://doi.org/10.1016/0076-6879(93)21011-V
State	Published - Jan 1 1993
Externally published	Yes

ASJC Scopus subject areas

Biochemistry
Molecular Biology

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10.1016/0076-6879(93)21011-V

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@article{d87137cbbb03474fb2082f65841c4713,

title = "[9] Visualization of Exocytosis by Quick Freezing and Freeze-Fracture",

abstract = "This chapter discusses the visualization of exocytosis by quick freezing and freeze-fracture. Exocytosis occurs extremely rapidly. Quick freezing in combination with freeze-fracture offers advantages for visualizing exocytosis in that it captures events occurring within milliseconds after stimulation and reveals panoramic views of membranes. Cells frozen ultrarapidly{\textemdash}that is, by removing heat so fast that ice crystals are undetectable by electron microscopy (EM), can yield superbly preserved ultrastructure without membrane fusion-mimicking artifacts resulting from slow, selective chemical fixatives and membrane-disrupting cryoprotectants. The aluminum planchet to which specimens are frozen is designed to fit directly onto the Balzers (Nashua, NH) 301 specimen table and is secured with a screw cap. Alternatively, tables fitted with a pair of flange clamps can be custom designed for many freeze-fracture units, including the Balzers 400 model.",

author = "Merkle, \{Carrie J.\} and Chandler, \{Douglas E.\}",

note = "Funding Information: concerning specimen stage configurationsf or cold metal block freezingm achines, Lisa Moore for bibliographica ssistance,a nd Charles Kazilek for photographic assistance.R esearch from our laboratory described herein was supported by National Science Foundation Grant IBN-9117509.",

year = "1993",

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doi = "10.1016/0076-6879(93)21011-V",

language = "English (US)",

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pages = "112--123",

journal = "Methods in Enzymology",

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T1 - [9] Visualization of Exocytosis by Quick Freezing and Freeze-Fracture

AU - Merkle, Carrie J.

AU - Chandler, Douglas E.

N1 - Funding Information: concerning specimen stage configurationsf or cold metal block freezingm achines, Lisa Moore for bibliographica ssistance,a nd Charles Kazilek for photographic assistance.R esearch from our laboratory described herein was supported by National Science Foundation Grant IBN-9117509.

PY - 1993/1/1

Y1 - 1993/1/1

N2 - This chapter discusses the visualization of exocytosis by quick freezing and freeze-fracture. Exocytosis occurs extremely rapidly. Quick freezing in combination with freeze-fracture offers advantages for visualizing exocytosis in that it captures events occurring within milliseconds after stimulation and reveals panoramic views of membranes. Cells frozen ultrarapidly—that is, by removing heat so fast that ice crystals are undetectable by electron microscopy (EM), can yield superbly preserved ultrastructure without membrane fusion-mimicking artifacts resulting from slow, selective chemical fixatives and membrane-disrupting cryoprotectants. The aluminum planchet to which specimens are frozen is designed to fit directly onto the Balzers (Nashua, NH) 301 specimen table and is secured with a screw cap. Alternatively, tables fitted with a pair of flange clamps can be custom designed for many freeze-fracture units, including the Balzers 400 model.

AB - This chapter discusses the visualization of exocytosis by quick freezing and freeze-fracture. Exocytosis occurs extremely rapidly. Quick freezing in combination with freeze-fracture offers advantages for visualizing exocytosis in that it captures events occurring within milliseconds after stimulation and reveals panoramic views of membranes. Cells frozen ultrarapidly—that is, by removing heat so fast that ice crystals are undetectable by electron microscopy (EM), can yield superbly preserved ultrastructure without membrane fusion-mimicking artifacts resulting from slow, selective chemical fixatives and membrane-disrupting cryoprotectants. The aluminum planchet to which specimens are frozen is designed to fit directly onto the Balzers (Nashua, NH) 301 specimen table and is secured with a screw cap. Alternatively, tables fitted with a pair of flange clamps can be custom designed for many freeze-fracture units, including the Balzers 400 model.

UR - https://www.scopus.com/pages/publications/0027160965

UR - https://www.scopus.com/pages/publications/0027160965#tab=citedBy

U2 - 10.1016/0076-6879(93)21011-V

DO - 10.1016/0076-6879(93)21011-V

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AN - SCOPUS:0027160965

SN - 0076-6879

VL - 221

SP - 112

EP - 123

JO - Methods in Enzymology

JF - Methods in Enzymology

IS - C

ER -

[9] Visualization of Exocytosis by Quick Freezing and Freeze-Fracture

Abstract

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